Basler Microscopy Software Owner's Manual

Basler Microscopy Software Owner's Manual
User Manual
Basler Microscopy
Software
Contents
1.
About your software..........................................................................................3
2.
Working with the Basler Microscopy Software...............................................4
2.1. Overview - User interface ........................................................................................................ 4
2.2. Working with documents ......................................................................................................... 5
3.
Acquiring images ..............................................................................................7
3.1. Acquiring a single image ......................................................................................................... 7
3.2. Camera Control ....................................................................................................................... 9
3.2.1.
3.2.2.
3.2.3.
3.2.4.
3.2.5.
Maximizing the live-image to screen .................................................................... 14
Automatic exposure time ...................................................................................... 14
Manual exposure time .......................................................................................... 16
Setting the camera resolution ............................................................................... 17
Toggle Subarray ................................................................................................... 18
3.3. Acquisition Settings ............................................................................................................... 20
3.3.1.
3.3.2.
3.3.3.
3.3.4.
3.3.5.
3.3.6.
3.3.7.
3.3.8.
4.
Acquisition Settings - Acquisition - General ......................................................... 20
Acquisition Settings - Document Name ................................................................ 22
Acquisition Settings - Saving ................................................................................ 23
Acquisition Settings - Camera - General .............................................................. 24
Acquisition Settings - Camera - Adjustment ......................................................... 26
Acquisition Settings - Camera - Color .................................................................. 27
Acquisition Settings - Camera - Exposure ............................................................ 28
Acquisition Settings - Camera - Information ......................................................... 28
Working with images.......................................................................................29
4.1. Changing the way an image is displayed .............................................................................. 29
4.2. Processing images ................................................................................................................ 32
4.3. Performing a white balance on an image .............................................................................. 33
4.4. Working with drawing objects ................................................................................................ 34
5.
Measuring images ...........................................................................................36
5.1. Overview................................................................................................................................ 36
5.2. Measuring images ................................................................................................................. 38
6.
Configuring the system ..................................................................................43
6.1. Overview................................................................................................................................ 43
6.2. Configuring the system.......................................................................................................... 44
6.3. Manual Magnification Calibration .......................................................................................... 45
About your software
1.
About your software
Acquiring images
Note: Different camera and acquisition settings are available depending on the
camera you are using.
Your system enables you to acquire high resolution images of a sample in just a
few steps. Your system is comprised of your software, a Basler camera, and your
microscope. With the image acquisition process, data is read out from the
camera that is attached to the microscope and is displayed on the computer's
monitor.
You can first observe the live-image and make good settings for it. The liveimage is constantly refreshed. This means that if you move the stage, the liveimage changes correspondingly. You can switch the live-image on and off and
acquire an image of the parts of the sample that interest you. When you do this,
you create a digital image that you can save and process or measure with a
variety of your software's functions.
Measuring images
You can make various measurements on images. You can measure the length of
a line for example. The measurement objects are displayed in the image's
drawing layer. All of the image information is therefore retained. The
measurement results are shown in a table and are saved together with the
image.
Processing images
You can process the images you acquired and optimize the image quality for
your purposes. Sharpen filters and contrast optimization functions are available
for this.
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Working with the Basler Microscopy Software
2.
Working with the Basler
Microscopy Software
2.1. Overview - User interface
The graphical user interface determines your software's appearance. It
determines which menus are shown, how the individual functions are called up,
how and where images are displayed, and much more. In the following, the basic
elements of the user interface are described.
Appearance of the user
interface
The illustration shows the schematic user interface with its basic elements.
(1) Menu bar
(2) Document group
(3) Toolbars
(4) Tool windows
(5) Status bar
(1) Menu bar
(2) Document group
(3) Toolbars
(4) Tool windows
(5) Status bar
You can call up many commands by using the corresponding menu. You can find
a variety of commands in the View menu with which you can change the
appearance of the user interface.
The document group contains all of the loaded images.
When you start your software, the document group is empty. While you use your
software it gets filled - e.g., when you load or acquire images, or perform various
image processing operations to change the source image and create a new one.
Commands you use frequently are linked to a button providing you with quick
and easy access to these functions. Please note, there are many functions which
are only accessible via a toolbar, the drawing functions required for annotating an
image for example.
Tool windows combine functions into groups. These may be very different
functions. For example, in the Properties tool window you will find all the
information available on the active image. In contrast to dialog boxes, tool
windows remain visible on the user interface as long as they are switched on.
That gives you access to the settings in the tool windows at all times.
The status bar contains information, a brief description of each function for
example. Position the mouse pointer on the name of a command or on a button
to display a brief description of a function.
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2.2. Working with documents
There are a number of ways to open, activate, save, or close images.
Saving images
You should always save important images immediately following their acquisition.
You can recognize images that have not been saved by the star icon after the
image's name.
There are a number of ways in which you can save images.
Automatic save
1.
To save an individual image, activate it in the document group. Then use
the File > Save As... command, or the [Ctrl + S] keyboard shortcut.
2.
Use the Documents tool window.
Select the desired document and use the Save command in the context
menu. For the selection of documents, the standard MS-Windows
conventions for multiple selection are valid.
3.
Use the Gallery tool window.
Select the desired document and use the Save command in the context
menu. For the selection of documents, the standard MS-Windows
conventions for multiple selection are valid.
1.
When you exit your software, all of the images that have not yet been
saved will be listed in the Unsaved Documents dialog box. This gives
you the chance to decide which images you still want to save.
2.
You can also configure your software in such a way that all images are
saved automatically after image acquisition. To do so, use the
Acquisition Settings > Saving dialog box. Here, you can also configure
your software in such a way that all images are automatically saved in a
database after the image acquisition.
Closing images
There are a number of ways in which you can close images in your software.
Closing all images
Closing an image
immediately
1.
Use the Documents tool window.
Select the desired images and use the Close command in the context
menu. The standard MS-Windows conventions for multiple selection
apply for the selection of images.
2.
To close a single image, activate the image in the document group and
use the File > Close command. Alternatively, you can click the button
with the cross [ x ]. You can find this button at the top right of the image's
tab's header next to the name of the image.
3.
Use the Gallery tool window.
Select the desired images and use the Close command in the context
menu. The standard MS-Windows conventions for multiple selection
apply for the selection of images.
To close all of the images that are loaded, use the Close All command or the [Ctrl
+ Alt + W] keyboard shortcut. You can find this command in the File menu, and in
both the Documents and the Gallery tool windows' context menu.
To close an image immediately without being prompted to save it, press the
[Shift] key while closing it. Data you have not saved will be lost.
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Working with the Basler Microscopy Software
Opening images
There are a number of ways in which you can open or load images.
1.
Use the File > Open... command.
2.
Drag the images you want directly out of the MS-Windows Explorer into
your software's document group.
Note: At the same time, up to 150 documents can be loaded in the document
group.
Generating a test
image
If you want to get used to your software, then sometimes any image suffices to
try out a function.
Press [Ctrl + Shift + Alt + T] to generate a color test image.
With the [Ctrl + Alt + T] shortcut, you can generate a test image that is made up
of 256 gray values.
Activating images in the document group
There are several ways to activate one of the images in the document group and
display it on your monitor.
1.
Use the Documents tool window. Click the desired image there.
2.
Use the Gallery tool window. Click the desired image there.
3.
Click the title of the desired image in the document group.
4.
Click the small arrow
at the top right of the document group to open a
list of all of the loaded images. Left click the image that you want to have
displayed on your monitor.
Note: You can choose whether you want to activate the images in the
Documents tool window and in the Gallery with a single or a double mouse click.
Use the Tools > Options... command. Select the Environment > General entry in
the tree view.
Select the Activate documents with single click check box. Then it's sufficient to
click once with the left mouse button to have the image you have chosen
displayed in the document group.
Clear the check box. Now you activate the image with a double click.
Attaching images to an e-mail
1.
Load the images you want to attach to your e-mail.
2.
Use the File > Send E-mail... command.
3.
Check whether all images you want to attach are selected.
4.
Click the Send button to generate an e-mail with the selected images
included as attachments.
5.
•
You will receive a warning message if the sum of file sizes of all
images exceeds the maximum permitted size.
•
A new e-mail form will be opened by your e-mail program. Your email program does not have to be already running for this to happen.
The e-mail contains all of the selected image files as attachments.
Enter the recipient’s address and your message and then send off your
e-mail.
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Acquiring images
3.
Acquiring images
3.1. Acquiring a single image
You can use your software to acquire high quality images of a sample in just a
few steps. For your first acquisition you should carry out these instructions step
for step. Then, when you later make other acquisitions, you will notice that for
similar types of sample many of the settings you made for the first acquisition can
be adopted without change.
1.
Use the View > Layout > Reset Current Layout command.
•
You can find the Microscope Control (1) toolbar at the upper edge of
the user interface, right below the menu bar.
To the right of the document group, you can find the Camera Control
(2) tool window.
Setting the
magnification
2.
On the Microscope Control toolbar, click the button with the objective that
you use for the image acquisition.
Switching on the liveimage
3.
In the Camera Control tool window, click the Live button.
•
4.
The live-image (3) will now be shown in the document group. A new
image document will automatically be created for the live-image.
Go to the position on the sample you are interested in.
Preferences in the Acquisition Settings
Use the acquisition settings to make default settings for the image acquisition.
5.
Open the Acquisition Settings dialog box. To do so, click the Acquisition
Settings button in the Camera Control tool window's toolbar.
6.
Select the Camera > General entry in the tree view.
From the Color temperature list, select a predefined color setting to
acquire the sample in particular light conditions. Select the color
temperature you want.
7.
Carry out a Gamma correction.
Select the Camera > Adjustment entry in the tree view.
Move the Gamma slide control to increase the image contrast either in
bright image areas or only in dark image areas.
8.
Close the Acquisition Settings dialog box with OK.
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Acquiring images
Settings in the Camera Control
Use the functions of the Camera Control tool window to set the image quality.
9.
Acquiring and saving
an image
Bring the sample into focus. The Focus Indicator toolbar is there for you
to use when you are focusing on your sample.
10.
Check the color reproduction. If necessary, carry out a white balance.
11.
Check the exposure time. You can either use the automatic exposure
time function, or enter the exposure time manually.
12.
Select the resolution you want.
13.
In the Camera Control tool window, click the Snap button.
•
14.
The acquired image will be shown in the document group.
Use the File > Save As... command to save the image. Use the TIF file
format.
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Acquiring images
3.2. Camera Control
Use the Camera Control tool window to acquire images. This tool window is
displayed on the right side of the user interface by default. Should this tool
window not be visible, use the View > Tool Windows > Camera Control
command to show it.
Note: Some of the settings in the Camera Control tool window are camera
specific (e.g., the available resolutions). For this reason, your camera's
functionality can differ from what is described here.
Structure of the tool
window
The tool window is made up of several parts.
(1) Toolbar of the tool window
(2) Acquiring images
(3) Adjusting the exposure time
(4) Setting the resolution
(5) Entering an image comment
(6) Changing the camera settings
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Acquiring images
(1) Toolbar of the tool window
In the Camera Control tool window, you can find the following buttons. You can
find these buttons in a toolbar, at the top of the tool window.
White Balance
White balance your camera on the live-image.
Focus Indicator
Click this button to make the Focus Indicator toolbar
appear. Use this toolbar as a tool to help you bring the
sample into focus.
Toggle RGB /
Grayscale Mode
You can select whether you acquire true-color images
or gray-value images with your camera.
Acquisition Settings
Clicking this button will enable you to change a variety
of settings for the image acquisition.
Adjust Display
Click this button to have its corresponding tool window
Adjust Display displayed. This tool window offers you
possibilities for changing the appearance of the current
image.
When the tool window is on display, this button appears
clicked. You can recognize this status by the button's
background color. To close this tool window again, click
the button with a cross
in the tool window's header.
Toggle Saturation
Indicator
Apply a pseudo color scheme in live mode.
Maximize to Screen
Click this button to see only the live image on your
monitor. The user interface will be completely faded out.
You can save all current camera settings in the Camera
Load Camera Settings Control tool window as parameter sets and load them
again at any point in time. Use this when you want to
set certain defined acquisition conditions for specific
Save Camera Settings samples, for example.
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Acquiring images
White Balance
In live mode, click the small arrow next to the button to open a list of available
white balance functions and then select the function you want. The active
function is indicated by the icon on the button. The selected function is also
identified with a check mark in the menu.
Prerequisite: Your camera is operating in RGB mode.
Toggle RGB/Grayscale
mode
One Touch White
Balance
Select the One Touch White Balance command to
perform a white balance on the active live image. The
color settings that were determined are used until you
click the One Touch White Balance button again.
Automatic White
Balance
The color settings are continually checked and adjusted
in this mode.
White Balance on ROI
Use the White Balance on ROI command to define a
particular image segment that you want to use for the
white balance.
Some cameras can also be used to acquire gray-value images.
The Toggle RGB/Grayscale mode button looks like this when your camera is in
RGB mode and you are acquiring true-color images. Release this button to
acquire gray-value images with your camera.
The Toggle RGB/Grayscale mode button looks like this when your camera is in
grayscale mode and you are acquiring gray-value images. Click the button to
acquire color images with your camera.
Toggle Saturation
Indicator
Prerequisite: Your camera is operating in grayscale mode.
The saturation indicator tells you whether an image (or a part of an image) is
underexposed or overexposed. When the saturation indicator is switched on, all
of the pixels whose intensity value is too low are shown in blue and all pixels
whose intensity value is too high are shown in red in the live-image. If too many
blue pixels can be seen, the image is underexposed. If too many red pixels can
be seen, the image is overexposed.
The color table is only applied to the live-image. The image you acquire will be a
normal gray-value image.
(2) Acquiring images
Acquiring images
Click the Live button to switch your camera to the live mode. The live-image will
then be displayed in the image window. The button will change its appearance.
This enables you to immediately recognize that you are in the live mode.
When you are in the live mode, click the Live button once more to end this mode
without acquiring an image. You can change this behavior. To do so, use the
Acquisition Settings > Acquisition > General dialog box.
Click the Snapshot button to acquire a snapshot. The live mode will then, as a
rule, be exited. The acquired image will be shown in the document group.
Image acquisition several ways
You have several ways of switching to the live mode to acquire snapshots:
• Use the Camera Control tool window.
• Use either the Acquire > Live or the Acquire > Snap menu commands.
• Use the [F7] key to switch to the live mode.
Use the [F8] key to acquire an image.
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Acquiring images
(3) Adjusting the exposure time
Use the Exposure group, to set the exposure time.
The functions in the exposure time control depend on whether you choose to use
the automatic exposure time option or to set the exposure time manually. The
Automatic option is always set by default.
(1) Automatic exposure time
(2) Manual exposure time
Exposure time in the
image properties
The exposure time is adopted in the Properties tool window after each image
acquisition. You can find this information in the Camera group. This
information will be retained if the image is saved in the TIF file format.
(4) Setting the resolution
Use the Resolution group to set the camera resolution.
Note: Different resolutions are available, depending on the camera being used.
You can set different resolutions for the live-image and the acquired image.
Select the resolution you want from the Live list or the Snap list (1).
Another way of reducing your camera's resolution, is to reduce the size of the
image area (2) by using a subarray.
(1) Setting the camera resolution
(2) Toggle Subarray
(5) Entering an image comment
In the Camera Control tool window, you can enter text in the Note field. This text
will then be saved along with the image as a comment for every image
acquisition.
Note: The text in the Note field isn't saved when you save the current camera
settings. The text in the Note field is deleted when you close your software.
Viewing a comment
Displaying the
comment in the image
window
After the image has been acquired, you can view and edit the image comment in
the Properties tool window's Document > Note field. To do so, load the image,
and use, e.g., the View > Tool Windows > Properties command to make the tool
window appear.
You can have the image comment displayed in the image window, together with
the info stamp.
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Acquiring images
Deleting or editing
comments for individual
images
1.
Go to the Documents group, located in the Properties tool window.
•
2.
You will see that the comment you entered in the Camera Control
tool window has been adopted in the Note field.
Click in the Note field.
•
When the Note field is active, a small button with three dots will
appear.
3.
Click this button.
4.
Delete or change the comment in the Note dialog box.
(6) Changing the camera settings
Use the Camera Settings group to change brightness, contrast and the color
settings.
Brightness and
Contrast
Use the Brightness slide control, if the whole image is too bright or too dark.
Values smaller than 0 lower the image brightness. Values greater than 0
increase the image brightness.
Move the Contrast slide control to change the contrast values of the image.
Values smaller than 1 lower the image contrast. Values greater than 1 enhance
the image contrast.
When changing the image contrast the brightest part of the image always
remains the same.
After changing the image contrast it may be that dark or bright image areas are
not displayed optimally anymore. In this case use the Brightness slide control to
make the whole image brighter or darker.
You can also change the Contrast and Brightness values in the Acquisition
Settings > Camera > Adjustment dialog box.
The values are automatically saved together with the image. You can view the
values in the Properties tool window.
Hue and Saturation
Increase the Saturation value when there is a preponderance of one color in your
images. In this way you will attain a better reproduction of the other colors in your
image.
To change the parameter, move the slide control. A change made in the
Saturation will have an effect on all of the color channels.
Use the Hue value to alienate the color impression of the acquired images.
Certain structures in the image can thus be better recognized, for example.
A value of 180 means that the color in the image corresponds to the color of the
sample.
You can also change the Saturation and Hue values in the Acquisition Settings >
Camera > Color dialog box.
The values are automatically saved together with the image. You can view the
values in the Properties tool window.
Live frame rate
The Live frame rate field shows the current frame rate (images per seconds) in
the live mode.
In the live mode, a huge amount of data is transferred from the camera to your
PC. It may occur that the USB connection to your system is not sufficiently high
to support the full data rate. This may lead to certain errors in the live-image, for
example, stuttering or color distortion.
In this case, use the Data rate slider to reduce the amount of data that is
transferred. You can find this slider in the Acquisition Settings > Camera >
Adjustment dialog box.
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Acquiring images
3.2.1.
Maximizing the live-image to screen
To see only the live image on your monitor, click the Maximize to Screen button.
The user interface will be completely faded out. You can find this button in the
Camera Control tool window.
Size of the live-image
The live-image's zoom factor will not be changed when you switch to the
Maximize mode. If you want to see the whole of the live-image in Maximize
mode, click the Fit to Window button before you switch to the Maximize mode.
Switching off the
Maximize mode
To switch off the mode and return to the normal user interface, use the [Esc] key
or click the Exit Maximized Screen button.
Setting the image
background
In the Maximize mode, the live-image does not necessarily take up all of your
monitor's screen. You can set a different color for the background. When you are
working with a fluorescence acquisition, for example, the diffused light can cause
a disturbance when you are looking at an image with a gray background. In this
case you can set a black background.
To do this, use the Tools > Options... command. Select the Images > General
entry in the tree view. Select the background you want from the Background list.
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3.2.2.
Automatic exposure time
Some of the exposure time correction settings depend on the model of camera
that you are using. For this reason, the Exposure group might have a somewhat
different appearance for your camera.
Switching on the
automatic exposure
time
Functions in the
"Exposure" group
The functions for setting the exposure time can be found in the Camera Control
tool window.
Select the Automatic option located in the Exposure group to choose an
automatic exposure time.
In the automatic exposure time mode, your system will continuously calculate the
optimum exposure time for the live-image. The snapshot acquisition always uses
the last exposure time that was calculated in the automatic exposure time mode.
The Exposure group in the Camera Control tool window offers options for
effecting the functioning of the automatic exposure time mode.
(1) Displaying the exposure time
(2) Locking the current exposure time
(3) Setting the region
(4) Setting the gain
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Acquiring images
(1) Displaying the exposure time
The current exposure time is displayed in the Camera Control tool window and
on the Camera Control toolbar.
The current exposure time is also shown in the function group's header in the
Camera Control tool window. This ensures that the exposure time is always
displayed, even if you minimize the function group and only the header remains
visible.
(2) Locking the current exposure time
Click the Lock Current Exposure Time
button to have the currently calculated
exposure time used for the complete duration of the live-acquisition.
In this way you will switch off the automatic adjustment for the duration of the
acquisition. That means that even when you move to a completely different place
on your sample, the exposure time will not be adjusted accordingly.
The Lock Current Exposure Time button remains active (also for later liveimages) until you switch it off again.
(3) Setting the region
The exposure time is always calculated on the basis of the histogram, that is, the
intensity distribution in the image. From the Region list, select the image segment
that is to be used for the calculation of the histogram and thus of the automatic
exposure time.
Full Image
Spot
Select the Full Image option so that the histogram of the entire image goes into
the calculation of the exposure time. This method is recommended if the
structures to be observed are equally dispersed throughout the entire image.
Select the Spot option to have the histogram of only a rectangular image
segment calculated. The sizes shown in the Region list depend on the camera
model being used. As soon as you switch to the live-image you will see a white
dashed rectangle that identifies the relevant segment of the image. With some
cameras you can move the rectangle to any position in the image by dragging it
there with your left mouse button pressed, and in this way you will then influence
the exposure time. You can use this method if relevant details are only visible in
a part of the image. The appearance can thus be optimized for precisely this
segment of the image.
You can also change the image segment you've selected, however you want to,
at a later date.
(4) Setting the gain
Use the Gain slide control when you want to decrease the exposure time without
making the image darker. The more you increase the gain, the shorter the
exposure time becomes. This increases the image noise, however. So, only use
the Gain slide control if it's especially important to have a short exposure time.
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Acquiring images
3.2.3.
Manual exposure time
Switching on the
manual exposure
The functions for setting the exposure time can be found in the Camera Control
tool window.
Select the Manual option. Then you can freely set the exposure time.
Functions in the
"Exposure" group
(1) Setting and displaying the exposure time
(2) Setting the gain
(1) Setting and displaying the exposure time
Adjusting the exposure
time
Displaying the
exposure time
There are several different ways of manually setting the exposure time.
•
To do this, move the slide controls.
•
Click the [-] and [+] buttons to alter the exposure time in small steps.
•
Enter an exposure time in the edit field, then press the [Enter] key. Or
change the exposure time by using the arrow buttons next to the edit
field.
The current exposure time is displayed in the Camera Control tool window and
on the Camera Control toolbar.
The current exposure time is also shown in the function group's header in the
Camera Control tool window. This ensures that the exposure time is always
displayed, even if you minimize the function group and only the header remains
visible.
(2) Setting the gain
Use the Gain slide control when you want to increase the sensitivity without
changing the microscope's exposure time or illumination intensity. The more you
increase the gain, the brighter the image becomes. It makes sense to use this
setting when the fluorescence is weak, for example.
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Acquiring images
3.2.4.
Setting the camera resolution
In the Camera Control tool window, you can use the Resolution group to set the
camera's resolution. Should this group be hidden, just double click on the
Resolution area.
Note: Most cameras can acquire images in various resolutions. Which
resolutions are possible, depends on the camera being used.
Functions in the
"Resolution" group
(1) Setting the resolution for a live image
(2) Setting the resolution for a snapshot
(3) Using the same resolution for live and snapshot
(4) Toggle Subarray
(1) Setting the resolution for a live image
Select the value you want for the live-image's resolution, in the Live list. A
resolution of 1920x1080 means, for example, that the image you acquire will be
made up of 1920x1080=2073600 pixels.
With most cameras you can reduce the resolution in the live-image. The method
will be specified in brackets behind the resolution. The smaller the resolution you
select is, the quicker the live-image will react. Inversely, the spatial resolution in
the image will improve with the increase in resolution.
Binning
With binning, pixels on the camera will be combined when they are read out.
Then less data will be transferred from the camera into the software, and the liveimage will be updated more quickly. At the same time, the noise in the image will
decrease.
When you select a resolution using the binning mode the term 'binning' will be
added in brackets after the resolution.
Toggle Subarray
When you reduce the size of the image area the speed in the live-image will be
increased, just as it is with binning, and you reduce the size of the file. In contrast
to binning, however, when you reduce the image area, the image segment
displayed in the live-image will be displayed in the same spacial resolution, that's
to say, without loss of quality.
You can find the functions for reducing the image area, in the Resolution group.
(2) Setting the resolution for a snapshot
Select the value you want for the snapshot's resolution, in the Snapshot list.
If you acquire images with a lower resolution, it will provide you with files that are
smaller. When you use a camera resolution of 1920x1080 an image with the
highest resolution will need approx. 6000 KB of storage capacity, and the same
image with the lowest resolution around 500 KB.
After the image has been saved, its size will be shown in the Size on Disk field in
the Documents group in the Properties tool window.
Display of the
resolution
The current resolution for the image acquisition is also shown in the function
group's header in the Camera Control tool window. This ensures that the
resolution is always displayed, even if you minimize the function group and only
the header remains visible.
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Acquiring images
(3) Using the same resolution for live and snapshot
To use the same resolution for the live-image and the snapshot acquisition, click
the button with the chain icon. Then you only need to set another resolution in
one place.
From that moment on, the Live list will be grayed out. Then you can't set anything
there. The value shown will, however, be updated if you set another value in the
Snapshot field.
(4) Toggle Subarray
Trim the image that your camera acquires.
When does which resolution make sense?
High resolution
In the highest resolution even sample structures that are very near to each other
are still shown separately in an image. This image contains the most information
on the sample. The disadvantage is, that a high resolution reduces the speed at
which the live-image is acquired.
Low resolution for liveimage
If you want to increase the live-image's reaction speed, it can make sense to set
a lower resolution. Then less data will be transferred from the camera into the
software, and the live-image will be updated more quickly. Admittedly, a
reduction of the resolution will also always result in less image information being
acquired.
Different resolution for
live-image and
snapshot
Note: You can set a different (lower) resolution for the live-image than that which
you use for the snapshot. That makes sense, for example, when you wish to
have a live-image that reacts quickly, but do not want to accept a loss of image
quality when you acquire a snapshot.
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3.2.5.
Toggle Subarray
The Toggle Subarray button is located in the Camera Control tool window. You
can find the button in the Resolution group. With it you trim the image that your
camera acquires.
Why use the "Toggle Subarray" mode?
You can use the Toggle Subarray mode for a variety of purposes.
Quick live-image
With cameras that support this function, the Toggle Subarray mode will lead to a
quicker live-image acquisition. You define a rectangular image segment and the
readout for the live acquisition limits itself to this area. A higher frame rate is
attained because less data needs to be transferred. Please note that you have to
select the Apply to Live check box before you can use the Toggle Subarray mode
on the live-image.
Toggle Subarray Binning
Like with the Toggle Subarray mode, binning (decreasing the resolution)
increases the speed of the live-image.
As opposed to binning, with the Toggle Subarray mode you will in fact see only a
small sample segment, this sample segment can, however, be displayed with the
greatest possible camera resolution. Use the Toggle Subarray mode when you
need an active-image that reacts quickly for focusing, and you only want to
reproduce very small sample structures.
The binning functions can also be found in the Resolution group.
You can also combine binning with the Toggle Subarray mode. In this way you
will increase the speed in the live-image still further, and also reduce the size of
your files even more.
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Acquiring images
Trimming the image
Sometimes it can be useful to trim an image you have acquired. By using the
Toggle Subarray mode, you can already trim uninteresting borders from your
images when you acquire them, and in this way considerably reduce the size of
your files.
Should you not have set an area for the Toggle Subarray mode before you
acquired an image, but are only interested in a part of this image, you can trim it
at any time by using the Image > Crop command.
Description of the function
Switching "Toggle
Subarray" mode on or
off
Switch to the live mode. Click the Toggle Subarray button to switch on the Toggle
Subarray mode. The button will retain its clicked appearance. You can then
immediately see whether the Toggle Subarray mode is switched on or off.
Note: You can only switch the Toggle Subarray mode on and off in the liveimage.
Using the "Toggle
Subarray" mode for the
live-image too
Specifying the image
segment for the
"Toggle Subarray"
mode
The Toggle Subarray mode will, in every case, have an effect on the snapshot
acquisition. Should you also want to use the Toggle Subarray mode with the liveimage, select the Apply to live check box.
When the Toggle Subarray mode is switched on, a blue frame in the image
window will show the active image segment.
You can at any time change the image segment that has been selected in the
live-image with the help of your mouse.
Note: The image segment for the Toggle Subarray mode can only be changed
when the Apply to live check box has not been selected. This is because, when
the Toggle Subarray mode has been activated, only a part of the image will be on
display, which means it would only be possible to define a new image segment
within the segment on display, but not outside this segment.
Specifying the image segment for the "Toggle
Subarray" mode
1.
Make the Camera Control tool window appear. To do this, you can use
the View > Tool Windows > Camera Control command.
2.
Switch to the live mode. To do this, click the Live button.
3.
If necessary, clear the Apply to Live check box in the Resolution group.
4.
Switch on the Toggle Subarray mode. To do so, click the Toggle
Subarray button.
•
In the image window a blue frame will then identify the active image
segment for the Toggle Subarray mode. If you can't see the frame,
click the Toggle Subarray button once more.
5.
To change its position, with the left mouse button depressed, take the
frame and drag it into the position you want. Position the image segment
for the Toggle Subarray method over an important image detail.
To change its size, move your mouse pointer over one of its handles. By
dragging the marker with the mouse button depressed, you can adjust
the frame's size as wished.
6.
Click the Snap button to stop the live mode and thus acquire a new
image.
•
The acquired image will now only show the image segment you have
chosen.
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Acquiring images
3.3. Acquisition Settings
Use the Acquisition Settings dialog box to change a variety of settings for the
image acquisition.
Opening the dialog box
To open this dialog box, click the Acquisition Settings button. You can find this
button on the Camera Control toolbar and on the Camera Control tool window's
toolbar.
The dialog box's tree view offers you access to dialog boxes with a variety of
setting options. Select an entry in the tree view to display the respective dialog
box.
Note: The exact functionality in the camera settings depends on the camera
used. For this reason, the functionality in your software may differ from that
described here.
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3.3.1.
Acquisition Settings - Acquisition - General
This dialog box offers you several general options for working with live images.
Live
The live-image will be allocated its own window in the document group. This
window's title will be Live (active). The behavior of this live window depends on
the settings in the Live group.
Closing the live window
when stopping the liveimage
Select the Close document when Live is stopped option to have the live window
closed every time you exit the live mode.
When you exit the live mode by clicking the Live button, in this case, no image
will be acquired.
To acquire an image, exit the live mode by clicking the Snap button.
Keeping the live
window when stopping
the live-image
Select the Keep document when Live is stopped option to keep the live-image
when you exit the live mode.
When you exit the live mode by clicking the Snap button, the live-image will be
stopped and additionally an image acquired. You will find the stopped live -image
in the image window with the title Live (stopped). You can save the stopped liveimage just as you can every other image.
Note: A stopped live-image is not the same image as the acquired snapshot. This
means that you can set different resolutions for the live-image and the snapshot
acquisition. In that case, a stopped live-image and an acquired individual image
each have different resolutions.
When you exit the live mode by clicking the Live button, the behavior in this
mode depends on the status of the Create new document when Live is started
check box.
Creating a new
document when the live
mode is started
The Create new document when Live is started check box is only relevant when
you've selected the Keep document when Live is stopped option.
Clear the Create new document when Live is started check box to have only one
single live window.
When you exit the live mode by clicking the Live button, you will find the stopped
live-image in the Live (stopped) live window. The next time you start the live
mode, the contents of the live window will be overwritten.
Select the Create new document when Live is started check box, if you want to
have a new window opened every time you start the live mode. In this way you'll
20
Acquiring images
prevent the overwriting of the last live window's contents.
If you finish the live mode now, the stopped live-image is kept in all cases. A new
image document will be created for the stopped live-image.
You can use this setting (Keep document when Live is stopped together with
Create new document when live is started) when you want to make a
measurement in the live-image. When you have finished making your
measurements, stop the live-image. You can then save the image you have
measured along with the measurements. When you start the next live-image, you
can perform a new measurement.
Continuing the live
mode after a snapshot
Select the Continue live after acquisition check box to only make a pause in the
live mode while you acquire a snapshot. A snapshot acquisition will then create a
new image window, but the window for the live-image will remain active and will
immediately switch back into the live mode.
To exit the live mode, click the Live button, located in the Camera Control tool
window.
Calibration
Choosing the basic unit
You can set the basic unit for the X/Y-calibration that is to be used for the image
acquisition. To do so, select the unit you want to use from the Basic Unit list. As
basic units Meters [m] and Inches [in] are available.
When you select another basic unit, all of the images that you from that moment
on acquire will be automatically calibrated in this new basic unit. Now, all values
that apply to the X/Y-calibration will be specified in this new basic unit. These
could be:
•
•
•
•
the labeling of the scale bar
the calibration data in the Properties tool window
the measurement results when you make measurements on an image
the label for the ruler that you can shown in the image window. Use the
Options > Images > View dialog box to show or hide rulers. The ruler's
label doesn't change until you restart your software.
Note: The basic unit for the X/Y-calibration of images you have already acquired
will not be changed, when you alter the basic unit. If you have acquired an image
with the basic unit Meters it will remain calibrated in meters or in a unit derived
therefrom, such as mm or µm. If you wish to measure this image in another basic
unit, use the Image > Calibrate Image... command to alter the basic unit.
Confirm after acquisition
Select the Confirm magnification after acquisition check box to have the Calibrate
Image dialog box automatically opened after every image acquisition.
Use this feature whenever you want to have the possibility to still change the
objective magnification for the image acquisition after switching to the live-image.
In the Calibrate Image > XY-Calibration dialog box, select the Magnification
option. Now, you can choose the objective magnification you have acquired the
image with from the Set magnification > Objective list. This ensures that the
image is properly calibrated.
When you clear the check box you have to select the current objective
magnification, e.g., by clicking the corresponding button on the Microscope
Control toolbar, before you start the image acquisition. Only then is the image
you acquire correctly calibrated.
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Acquiring images
3.3.2.
Acquisition Settings - Document Name
When an image is acquired, your software allocates it a default name. For
instance, the first image that your software acquires will be named Image_01, by
default. You can change this name in the Acquisition Settings > Document Name
dialog box.
Preview
The Preview field shows the name of the next image that you acquire. This
preview will be updated as soon as you change the image's name.
Customize
An automatically created name is made up of different parts. In the Customize
group, you define a prefix and specify the numbering system.
Defining a prefix
Enter the first part of the image's name in the Text field. Image is the default
prefix. By default, the acquired images are called Image01, Image02 ... .
If possible don't use any special characters for the image name. Certain special
characters, e.g. ? are not accepted when you enter them.
Shifting the prefix
You can also move the prefix that you defined in the Text field to any other
position in the name of the image.
1.
2.
Defining counters
To do this, click the All Options... button.
•
A dialog box will open in which you can assemble an automatically
created name from various placeholders.
•
You can find the prefix defined in the Text field in the top position of
the Selected properties column.
Select the prefix and use the Up and Down buttons.
In the Counter digits field, define how many digits the numbering should consist
of, e.g., 3 for the number 001. Please note that the value entered in the Counter
digits field will not place an upwards limit on the numbering. This means that if
you have entered a value of 2, for example, and the last image you acquired was
Image_99, the next image will be called Image_100.
If you want to start the numbering from a certain value, change the value in the
Counter start field. You can, for example, return the numbering to 1 if you have
acquired a great many images. Or you can continue the numbering of a series of
images from the previous day. If you change the value in the Counter start field
then the next image will always start with the number that has been set. All
additional images will be counted up by the value 1.
Each time your software restarts, the numbering of the images will start with the
value set in the Counter start field.
You can only change the value in the Counter start field if you have cleared the
Reset automatically check box.
Note: The value in the Counter start field may be ignored if automatic saving is
active. In this case, your software checks whether a file with the name wanted
already exists in the current directory. Should this file exist, the next higher
available number will automatically be used. To switch the automatic storage
process on or off, use the Acquisition Settings > Saving dialog box.
Automatically resetting
the counter
Select the Reset automatically check box when you want to be sure that images
that belong together will be consecutively, serially numbered without any breaks.
You can then no longer change the numbering of the images manually.
Customizing document
names
Your software supplies you with a number of placeholders that you can use in
image names. Click the All Options... button to choose the placeholders that you
want.
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Acquiring images
3041 30092013
3.3.3.
Acquisition Settings - Saving
By default, when you make an image acquisition, a new image document will be
created and displayed in the document group. You can rename and save this
image. If you have not already saved it when you end your software, you will be
asked if you want to do so. In the Acquisition Settings > Saving dialog box, you
can also have the image automatically saved after the acquisition.
Automatic save
You can automatically save all images after acquisition.
Automatically saving
images after acquisition
Select the File system entry from the Destination list if you want to save your
images as files. Now, in the Directory group, define the destination path for
saving your documents.
Note: At the same time, up to 150 documents can be loaded in the document
group. When you're acquiring an image, you receive an error message if the
maximum possible number of documents is already loaded. When you have
switched on automatic saving, the image is acquired and saved to the current
directory even though it can no longer be displayed in the document window.
Deactivating automatic
saving
File type
Select the No automatic save entry to switch off the automatic saving of images.
You will then have to save the images yourself by using e.g., the File > Save
As... command, when you have acquired an image and want to keep the image
file. In this case, no other functions will be available in this dialog box.
In the File type list, select the file format in which the images should be saved
after the acquisition. For the image file formats TIF and JPEG, there are also
additional settings that should be taken into account when you save images.
Click the Options... button if you want to see these settings or change them.
Note: Images acquired with your software always contain a range of additional
information which can be seen in the Properties tool window. This additional
information will only be retained if the images are saved in the TIF format.
Close after save
Select the Close after save check box to have the image document close
immediately after the image has been saved. The images will then be saved as
files. Please note that you then can't see the images in your software after the
image acquisition. Use this possibility to avoid taking up too much of your PC's
memory capacity when you acquire images.
Directory
Prerequisite: The Directory group is only available if you select the File system
entry from the Automatic save > Destination list.
Selecting a path
Creating a subdirectory
The Path field shows the directory that will currently be used when your images
are automatically saved.
Click the [...] button next to the Path field to choose a different directory into
which the images are to be saved after their acquisition.
When you save images automatically after the image acquisition, you have the
possibility of saving images that belong together in their own individual directory.
By default, all of the images that you acquire in one day will then be saved in one
separate directory. On the following day, a new directory will be automatically
opened. This enables you to always have a clear overview, even when you
acquire a great number of images.
Select the Create Subdirectory check box to have the acquired images saved in
their own subdirectory.
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Acquiring images
Click the Customize... button if you want to change the name that has been
suggested for the subdirectory. The Customize Subdirectory Name dialog box
opens.
Your software supplies you with a number of placeholders that you can use in
directory names. With your choice of the placeholder you also determine the
criterion with which your directory tree structure will be organized. This enables
you to set up a subdirectory for each user, for instance.
Checking the preview
The Preview field shows you the current subdirectory for the next image
acquisition. This preview will be updated as soon as you change the
subdirectory's name.
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3.3.4.
Acquisition Settings - Camera - General
Use this dialog box to make a variety of general camera settings. For example,
you can mirror the acquired image, use a color camera in black & white mode, or
select a color table.
Note: Some of the settings are camera specific. For this reason, your camera's
functionality can differ from what is described here.
Image type
Setting the bit depth
Some cameras can acquire images of different image types. From the Bit depth
list, select the type of image that you want to acquire with your camera. The
image types differ basically in their depth of bit, i.e., the amount of intensity
values that each pixel can adopt.
Which entries are available in the Bit depth list depends on your camera.
Select the 8-bit RGB color entry to acquire 24-bit true-color images.
You can use some cameras in a black & white mode. Select the 8-bit grayscale
entry to acquire 8-bit gray-value images.
Switching between the
color and grayscale
modes
Choosing the color
temperature
If you have to frequently switch between the color and the grayscale mode, use
the Toggle RGB/Gray Scale Mode button. You can find this button on the
Camera Control tool window's toolbar. You can then switch from one mode to the
other with a single mouse click.
A camera can only reproduce correctly colored images when it knows the
prevailing color temperature. When you use this process, the individual colors in
the image (Red, Green, Blue) will be scaled in such a way that the white area of
the image is correctly reproduced as white.
From the Color temperature list, select a predefined color setting to acquire the
sample in particular light conditions. This automatically chooses the suitable color
setting for your camera.
5000 K corresponds to morning or evening sunlight.
6500 K corresponds to overcast daylight.
2800 K corresponds to tungsten light.
Select the Off entry if you do not need to adjust the color settings to particular
light conditions. That makes sense, for example, when you wish to quantitatively
compare images that were acquired under different light conditions. The color
impression becomes considerably paler.
Note: You can change the settings for the color temperature directly in the live
mode. You can thus see what effect a change has on the color reproduction
straight away.
Use the white balance to adjust these default settings, if necessary.
Different methods for determining the white balance are available in the Camera
Control tool window, depending on the camera that you are using.
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Acquiring images
Mirror
Horizontal
Vertical
Horizontal and Vertical
Select the Horizontal check box to mirror the image horizontally during
acquisition. The illustration shows left the source image and right the horizontally
mirrored image.
Select the Vertical check box to mirror the image vertically during acquisition.
The illustration shows left the source image and right the vertically mirrored
image.
The two forms of mirroring can be combined with each other. The illustration
shows left the source image and right the horizontally and vertically mirrored
image.
Pseudo Color
Prerequisite: The Pseudo color group is only available if you selected the 8-bit
grayscale entry in the Bit depth list.
You can use color tables to have images displayed in color on your monitor.
Then every intensity value in an image will be allotted a hue.
Using no pseudo colors
Using the saturation
indicator
By default, when you acquire images, no pseudo color tables will be used. In this
case, the Pseudo Color > Off option will be selected.
Select the Use saturation indicator option to have all of the pixels with a low
intensity value displayed in blue, and all of those with a high intensity value
displayed in red. This color table being used now only applies to the live-image.
When you acquire this image, it will show all of the pixels back in their actual
color, in the document group.
With this pseudo color table you can immediately see if the image is well
illuminated. When there are both blue and red pixels in the image, the camera's
complete dynamic range will be in use. If too many blue pixels can be seen, the
image is underexposed. If too many red pixels can be seen, the image is
overexposed.
If you switch the Use saturation indicator option on and off frequently, use the
Toggle Saturation Indicator button. You can find this button on the Camera
Control tool window's toolbar. This allows you to switch the option on and off with
a single click of the mouse.
Using a pseudo color
table for gray-value
images
You can apply a color table at as early as the image acquisition. To do this,
select the Use pseudo color table for grayscale images option.
Selecting the pseudo
color table
Click the Pseudo color... button to change the pseudo color table you have
chosen, or to define a new one.
You can color only the live-image, or only the acquired image, or both, with the
pseudo color table you choose. To do so, select the one of the following entries
For live only, For snapshot only or For live and snapshot from the list.
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Acquiring images
3.3.5.
Acquisition Settings - Camera - Adjustment
Use this dialog box to manually change settings on the camera that affect the
image quality.
Note: Some of the settings are camera specific. For this reason, your camera's
functionality can differ from what is described here.
Changing the image
contrast in the liveimage
Before you change the settings, you should switch to the live-image, so that you
can immediately see how it changes. To do this, use the Acquire > Live
command.
Enhancement
Gamma
Move the Gamma slide control to increase the image contrast either in bright
image areas or only in dark image areas.
When each image acquisition is made, the Gamma value will be adopted in the
Properties tool window. This information will be retained if the image is saved in
the TIF file format.
You will change the parameters for all color channels simultaneously. If you want
to accentuate or tone down one particular color, use the functions in the
Acquisition Settings > Camera > Color dialog box.
Contrast and
Brightness
Move the Contrast slide control to change the contrast values of the image.
Values smaller than 1 lower the image contrast. Values greater than 1 enhance
the image contrast.
When changing the image contrast the brightest part of the image always
remains the same.
After changing the image contrast it may be that dark or bright image areas are
not displayed optimally anymore. In this case use the Brightness slide control to
make the whole image brighter or darker.
Use the Brightness slide control, if the whole image is too bright or too dark.
Values smaller than 0 lower the image brightness. Values greater than 0
increase the image brightness.
You can also view the Contrast and Brightness values in the Camera Control tool
window and you can also change them there.
The values are automatically saved together with the image. You can view the
values in the Properties tool window.
Limit throughput
In the live mode, a huge amount of data is transferred from the camera to your
PC. It may occur that the USB connection to your system is not sufficiently high
to support the full data rate. This may lead to certain errors in the live-image, for
example, stuttering or color distortion.
Note: You should only reduce the data rate if errors in the live mode occur on
your system.
In this case, use the Data rate slider to reduce the amount of data that is
transferred. In the Camera Control tool window, a correspondingly smaller frame
rate is now shown in the live mode.
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Acquiring images
3.3.6.
Acquisition Settings - Camera - Color
Use this dialog box to change the camera's color settings manually.
Note: Some of the settings are camera specific. For this reason, your camera's
functionality can differ from what is described here.
Changing the color
settings in the liveimage
Before you change the color settings, you should switch to the live-image, so that
you can immediately see how it changes. To do this, use the Acquire > Live
command.
Description of dialog box
White Balance
Prerequisite: You can only change the color settings when you are working in
your camera's color mode.
With the gain value, you can weight the individual color channels separately.
Increase, e.g. the R value, to increase the amount of red in the image.
You shouldn't usually change the gain value manually. Instead, use your
camera's white balance functions. The result of the white balance will be
automatically adopted in the fields of this dialog box.
You can change the gain individually for each color channel. To change a
parameter, move the slide control. R, G and B stand for the Red, Green and Blue
color channels.
The gain values being used are transferred to the Properties tool window with
every image acquisition. You will find the Gain (White Balance) entry in the
Camera group. This information will be retained if the image is saved in the TIF
file format.
Device dependent
white balance
The white balance is influenced by every optical component within the light path.
In particular, the objectives can affect the color impression of an image.
Select the Device dependent white balance check box to automatically save
each white balance performed. When you acquire images later on using an
objective with a valid white balance, this white balance will automatically be
loaded and applied.
If you have cleared the Device dependent white balance check box the latest
white balance will be used, regardless of the objective used.
Saturation and Hue
The sliders Saturation and Hue offer additional possibilities to influence the color
settings of your camera. Switch your system to the live mode before you open
the Acquisition Settings dialog box. When you change the saturation and hue
value now, you can observe directly in the image window how the color
reproduction is influenced.
Increase the Saturation value when there is a preponderance of one color in your
images. In this way you will attain a better reproduction of the other colors in your
image. To change the parameter, move the slide control. A change made in the
Saturation will have an effect on all of the color channels.
Use the Hue value to alienate the color impression of the acquired images.
Certain structures in the image can thus be better recognized, for example. A
value of 180 means that the color in the image corresponds to the color of the
sample.
You can also view the Saturation and Hue values in the Camera Control tool
window and you can also change them there. The values are automatically
saved together with the image. You can view the values in the Properties tool
window.
Default
Click the Default button to select the preset color settings. The current settings
will then be lost.
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Acquiring images
3.3.7.
Acquisition Settings - Camera - Exposure
Use this dialog box to set the maximum possible exposure times for the image
acquisition. By doing this you can prevent the exposure times from becoming
unnecessarily long.
Maximum exposure time
You can set the maximum possible exposure time for the live-image and for the
snap. To do this, use the slide controls. Alternatively, you can also enter the
value you want directly in the text field. The maximum value that you can enter
here depends on the camera you use.
The Manual and Automatic slide controls correspond to the two possible modes
for determining the exposure time, the automatic and the manual mode. You can
determine which mode you want to use in the Camera Control tool window, for
example.
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3.3.8.
Acquisition Settings - Camera - Information
Use this dialog box to look at data pertaining to your camera.
Displayed information
Camera name
The type of camera being used
Serial number
The camera's serial number
API version
Firmware version
Driver version
Information about the camera driver being used
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Working with images
4.
Working with images
4.1. Changing the way an image is displayed
An image can be displayed in very different ways in your software's image
window. There are several ways in which you can change the way your image is
displayed.
Enlarging or reducing the size of the image in the
image window
There are several different ways to change your image's zoom factor in the
image window. The current zoom factor is displayed on the bottom right in the
status bar.
On the left, the whole image is displayed in the image window. On the right, the
zoom factor has enlarged the image segment so that it can be viewed in higher
resolution. The scale bar relates to the magnification of the image in the image
window and is adjusted accordingly.
The different zoom options are described in the text that follows.
1.
Use the buttons on the Zoom toolbar.
2.
Use the Zoom Tool button on the Toolbox toolbar.
3.
If you are not in zoom mode: Right click on an image window. In the
context menu you can find several commands with which you can alter
the image's zoom factor.
4.
Rotate the mouse wheel to change the zoom factor.
•
5.
You can set whether you want to enlarge or reduce the size of the
image in the image window when you move the mouse wheel
forwards. To do this, check or clear the Zoom in the image by
scrolling the mouse wheel away from you (forward motion) check
box. You can find this check box in the Tools > Options > Images >
Zoom dialog box.
Use the Image Navigator tool window.
•
In the Image Navigator tool window, while keeping the left mouse
button depressed, drag the navigation frame to a smaller size. As
soon as you release the mouse button, only the image segment you
have selected will be shown in the image window.
•
Enter the zoom factor you want directly in the edit field below the
image area in the Image Navigator tool window and press the [Enter]
key, or use the slide control.
29
Working with images
Changing an image's color mapping
You can display gray-value images in color. You can do this using color tables.
Task
Create a color table which displays dark gray-values in yellow, and light grayvalues in different shades of red. And, the lighter the gray values are; the lighter
the shades of red are to be.
On the left is the gray-value image. On the right the image has been colored
with a color table. The coloring allows you to see more details in the dark and
light gray-value range.
Creating a color table
1.
Load a gray-value image. You can use the [Ctrl + Alt + T] shortcut to
create a test image.
2.
Use the Image > Pseudo color... command to open the Pseudo color
dialog box.
•
3.
Move your mouse pointer onto the color table in the dialog box.
•
4.
The current color table is shown in the Current Color Table group.
Your mouse pointer then takes on the form of a hand.
While keeping the left mouse button depressed, drag the mouse over the
lower gray values in the color table.
•
A blue bar under the color table indicates the gray-value range that
has been selected. You can now assign this gray-value range a
color.
5.
Click the left color field in the Edit Color Table group once.
6.
Select the required color, yellow for example, in the Colors dialog box
and confirm it with OK.
7.
Click the right color field in the Edit Color Table group once.
8.
Select the required color, yellow for example, in the Colors dialog box
and confirm it with OK.
9.
Click the Apply button in the Edit Color Table group.
10.
•
All of the current image's gray values that are within the selected
gray-value range will then be displayed in yellow.
•
The current color table will be correspondingly updated.
While keeping the left mouse button depressed, drag the mouse over the
upper gray values in the color table.
•
11.
A blue bar under the color table indicates the gray-value range that
has been selected. You can now assign this gray-value range a
color.
Click the left color field in the Edit Color Table group once.
30
Working with images
Saving a color table
12.
Select the required color, dark red for example, in the Colors dialog box
and confirm it with OK.
13.
Click the right color field in the Edit Color Table group once.
14.
Select the required color, white for example, in the Colors dialog box and
confirm it with OK.
15.
Click the Apply button in the Edit Color Table group.
16.
•
All of the current image's gray values that are within the selected
gray-value range will now be shown in either yellow or red. In
contrast to the yellow pixels, the red pixels become constantly
brighter, the higher their intensity value is.
•
The current color table will be correspondingly updated.
Click the Save Color Table button. You can find this button at the top of
the Pseudo Color tool window.
•
The Save Color Table As dialog box opens.
17.
Enter a descriptive name for the new color table in the Name field,
Yellow-Red for example.
18.
Click the Save button to save the color table and to return to the Pseudo
Color dialog box.
•
19.
You can apply the Yellow-Red color table to additional gray-value
images from now on.
Close the Pseudo Color dialog box with OK.
•
The active gray-value image is now displayed in color.
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Working with images
4.2. Processing images
The Process menu offers several image processing functions, with which you
can change an acquired image (e.g., increase the image contrast or the image
sharpness).
1.
Load the image you want to process, or activate the image in the
document group.
•
2.
Use one of the commands in the Process menu, e.g., Process >
Enhancements > Adjust Intensity... .
•
3.
4.
Please note that the Process menu will only be visible when an
image is loaded and active in the document group.
The image processing dialog box opens. The image processing
operation that is active will be shown in the dialog boxes header.
Click the small arrow next to the Preview button to open a list of all of the
preview functions. Select the Original and Preview entry.
•
This preview function displays the same image segment twice in the
dialog box. The first one shown is the source image. The second is
the image that results when the current parameters are used.
•
Most of the image processing operations need one or two of the
parameters that are shown in the Settings group.
Change the image processing operation's parameters. Decrease the
gamma value and increase the brightness, for example.
•
After every change that is made in a parameter, the operation will be
immediately applied to the source image, and the resulting image will
be shown in the preview window.
5.
Click the Default button, to readopt the preset parameters in the Settings
group, when the current parameter doesn't make sense to you.
6.
When you have found the optimal parameters, click the OK button to
have the active image processing operation applied to the image with the
active parameters.
•
The image processing dialog box will closed.
•
Please note that the image processing operation changes the source
image. No new image document will be created. You can, however
use the Edit > Undo command to restore the source image.
•
The new image is not automatically saved. The fact that a change
hasn't been saved is indicated by an asterisk shown behind the
image name in the document group.
The source image (left) has low contrast. Adjust the intensity to get a resulting
image (right) with significantly better contrast.
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32
Working with images
4.3. Performing a white balance on an image
Should an area in your image that ought to appear white look reddish, or is
another color, this means your image has a tinge. In such cases, use the
Process > Enhancement > White Balance... command to perform a white
balance.
1.
Load the image you want to process, or activate the image in the
document group.
2.
Use the Process > Enhancements > White Balance... command.
•
The Filter: White Balance dialog box opens.
3.
For the start, define a white point in the image. This should be a point or
surface that is actually white, but isn't displayed in white in the image.
4.
For example, click the Select white point using the pipette button.
5.
•
The dialog box disappears. The mouse pointer has turned into a
pipette.
•
The size of the pipette's point is predefined at 3x3 pixels.
Alternatively, you can also use the ellipse or the rectangle to define
the white point, and by doing so, define an arbitrarily large area as
the white point.
Click a position in the image that you want to define as the white point.
•
6.
7.
The position is then marked with a red cross. When that has
occurred, you can still move the point. To do this, move your mouse
over the red cross so that it takes on this shape. Then, while
pressing the left mouse button, you can move the white point
wherever you want to.
Confirm your choice by clicking your right mouse button, then using the
Confirm Input command in the context menu.
•
The Filter: White Balance dialog box reopens.
•
In the Interactive white-point definition group, the color values of the
three colors for the defined white point are displayed.
•
In the White-point correction factors group, the correction values
determined for each color, are displayed.
•
In the dialog box's preview image, you can see what the corrected
image will look like, should you adopt these values.
•
If necessary, you can still manually change the white point correction
factors that have been determined. The preview window will then be
immediately, correspondingly updated.
When you are satisfied with the preview image, click the Apply button,
then click OK.
•
The white balance will be performed immediately. This means that
the colors in the image are adjusted to make the selected white point
and all of the areas with the same color values appear as white. You
can see the results immediately in the image.
8. If you are not satisfied with the results: Please note that you can undo
the white balance by using the Edit > Undo command.
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Working with images
4.4. Working with drawing objects
Various drawing functions are available that you can label your images with.
Inserting drawing objects
Task
Inserting an arrow
You want to insert an arrow into an image and label it.
1.
Load the image that you want to label.
2.
Set the zoom factor of the image window to display the image in a size
that makes the label is easy to read. You can rotate the mouse wheel to
change the zoom factor in the image window.
3.
Use the View > Toolbars > Drawing command to have the Drawing
toolbar displayed.
4.
Click the Arrow button in the Drawing toolbar.
•
Inserting a text object
Showing and hiding
drawing objects
There is a button for each drawing object on the Drawing toolbar.
Click the button for the drawing object that you want to insert and
then define the drawing object on the image.
5.
Keep the left mouse button pressed to draw the arrow.
6.
Double click the arrow to open the Drawing Object Properties dialog box.
Make the settings for the arrow's color and line width here. The size of
the head of the arrow is automatically adjusted to the width of the line.
7.
Click the Text button on the Drawing toolbar.
8.
Keep the left mouse button pressed draw a rectangle. Enter the required
text and format it.
9.
Use the [Esc] key to leave text-entry mode.
10.
You can display the image and the drawing layer together, on your
monitor, or alternatively the image without the drawing layer. Click the
Show Drawings button in the Drawing toolbar, to switch between
showing and not showing the drawing layer.
Two drawing objects have been defined on the image: an arrow and a label for
the arrow.
Selecting drawing objects
Selecting several
objects
For certain actions it is useful to select more than one drawing object. This
enables you to copy several drawing objects from one image to another at the
same time, or to format several drawing objects simultaneously.
1.
Use the buttons in the Drawing toolbar to create several drawing objects.
2.
If necessary, switch to the edit-object mode. To do so, click the Select
Drawing Objects button on the Drawing toolbar. The button will appear
clicked when the edit-object mode is active. You can recognize this
34
Working with images
status by the button's colored background.
When the button already appears clicked, you will already be working in
the edit-object mode.
3.
Clearing the selection
Use one of the following methods to select a collection of drawing
objects.
•
Left click an object.
While keeping the [Shift] key depressed, click other objects you want
to edit.
Objects you have selected will be identified by white or gray markers.
•
While keeping the left mouse button depressed, drag a rectangle to
encompass several objects.
All objects that are completely inside this rectangle will be selected.
All of the other existing selections will be deleted.
•
To select all of the drawing objects on the image, use the [Ctrl + A]
shortcut.
4.
If necessary, switch to the edit-object mode. To do so, click the Select
Drawing Objects button on the Drawing toolbar.
If the button already appears clicked, this means that you are already in
the edit-object mode.
5.
There are several ways of clearing a selection of drawing objects:
•
Use the Edit > Deselect command.
•
Use the [Ctrl + D] keyboard shortcut.
•
Click any place in the background of a document with your left
mouse button. The complete current selection will be cleared.
•
To deselect individual objects, click an object that has already been
selected.
Determining the default formatting for drawing
objects
1.
Insert a drawing object and assign the properties to it that you want to
use as default settings. For example, change the line color from black to
red if you want the lines in your images to be red by default.
2.
If necessary, select the drawing object that you want to use as a
template for the formatting.
3.
Use the Draw > Set As Default For Drawings command. You can find the
Draw menu on the Drawing toolbar.
•
4.
The drawing object that has been selected is now used as a template
for all new drawing objects.
Insert another drawing object, e.g., an ellipse.
•
The new drawing object will then be formatted in exactly the way you
determined that the first drawing object should be.
35
Measuring images
5.
Measuring images
5.1. Overview
You can measure distances and areas with your software. All of the results are
saved together with the image.
Prerequisite
For making measurements, correctly calibrated images are an essential
prerequisite.
Images that you have acquired with your software will have been automatically
correctly calibrated when you have specified the objective you used.
Should the image not yet have been calibrated, use the Image > Calibrate
Image... command to carry out a calibration.
Selecting the measurement environment
Measuring with help of
the tool window
Use the View > Tool Windows > Measurement command. In the Measurement
tool window, you have fast access to the measurement functions and to the
settings which effect measurements. This tool window is at the same time the
measurement display and contains all of the values that have been measured on
the active image.
Starting a measurement
Begin a measurement by selecting the measurement function you want. You can
find the measurement function in the Measurement tool window, on the
Measurement toolbar, or in the Measure menu.
Working in the default
measurement mode
As soon as you have clicked a measurement function, your software will
automatically switch to measurement mode. You can measure image structures
in measurement mode. When you have finished making your measurements you
automatically leave measurement mode.
You can now carry out another measurement, or use any other software
functions that you want.
Working in continuous
measurement mode
Double click a measurement function to switch to continuous measurement
mode. In continuous measurement mode, your mouse pointer turns into a cross.
In the continuous measurement mode, you can immediately make further
measurements with these measurement functions. The continuous measurement
mode is valid for all loaded images. You can, therefore, easily measure
numerous images one after the other.
The button that has been selected in the Measurement toolbar remains active to
identify the current measurement function.
You will remain in this measurement mode until you explicitly switch it off. To do
so, click the Select Measurement Objects button on the Measurement toolbar.
Displaying and saving measurement results
The measurement results will be displayed directly on the image and in the
Measurement tool window. Should this tool window not be visible, use the View >
Tool Windows > Measurement command to display the tool window.
Saving the
measurement results
The measurements will be saved along with the image, if you save the image in
TIF format.
36
Measuring images
Editing measurements
You can edit existing measurement objects at any time. The measurement
values in the Measurement tool window will be correspondingly updated.
Note: When you load an image file that has measurement objects, it is only
possible to edit the measurement objects if the image file has been saved in the
TIF image file format.
Selecting measurement
objects
Before you can edit measurement objects, you have to select them. To do so,
click the Select Measurement Objects button, and then select the measurement
object(s). You can find the button both in the Measurement tool window and on
the toolbar.
Moving measurement
objects
You can move a whole measurement object while keeping the left mouse button
pressed.
Increasing/decreasing
the size of
measurement objects
You can also change the size of a measurement object. Move the pointer onto a
marker. By dragging the marker with the mouse button depressed, you can
adjust the frame's size as wished.
Change the measurement object by moving the handles.
Deleting measurement
objects
Click the [Del] key on your keyboard in order to delete the selected measurement
object. You can select measurement objects that you want to delete in the image
or in the table in the Measurement tool window.
Displaying helper lines
You can display helper lines for the Arbitrary Line measurement function. Select
a measurement object in an image and right click to open a context menu that
contains the Helper Lines command.
Measuring in the live mode
The measurement functions are also available in the live-image. You can
therefore, e.g., quickly measure a segment in the live-image.
Note: The measurements that you perform in the live-image aren't transferred to
the image when it is acquired.
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Measuring images
5.2. Measuring images
You can measure distances and areas with your software.
Measuring image objects interactively
Task
You want to measure the filaments in a superconductor.
To do this, load a suitable image, or acquire one. Measure the diameter of some
of the hexagonal filaments, between opposite corners. Then edit the
measurement and delete some of the measurements that have been performed.
1.
Use the View > Tool Windows > Measurement command to display the
Measurement tool window.
•
2.
Setting the labeling
color
You'll find the tool window (1) at the lower edge of the user interface.
Acquire an image or load one.
The measurement results will be written into the image according to the default
settings, in a red font and without a background. This can make the
measurement results hard to read on some images. Change the labeling
settings.
3.
Use the Tools > Options... command.
4.
Click the Measurement > Measurement Display entry in the tree view.
5.
Click in the Background Color field and choose a color, black for
example.
38
Measuring images
Measuring lengths
6.
Select the Text color > Fixed colors option and select a suitable color
from the palette. Select the color white to display the measurements in
white and the labels in white on a black background.
7.
Close the dialog box with OK.
8.
Click the Arbitrary Line button. You can find the button on the toolbar at
the top of the tool window.
9.
Click with your left mouse button at the starting point and end point of the
reference distance.
10.
Repeat the measurement several times.
11.
Take a look at the results in the tool window and in the image.
•
Deleting measurements
12.
Click one of the measurement results in the Measurement tool window.
•
13.
Closing the image
14.
The corresponding line will be selected in the image.
Press the [Del] key.
•
The measurement will be deleted both in the image and in the tool
window.
•
When a measurement has been deleted, the image and the tool
window contain one measurement less. The IDs of the remaining
measurements won't be changed by the deletion of a measurement.
Click the button with the cross [ x ] to the right of the image name in the
document group.
•
15.
The illustration shows the image with three executed measurements.
The measurement 2 has been selected.
You have changed the image because you've added interactive
measurements. For this reason, you'll receive a query whether you
wish to save the image or not.
Save the image in the TIF file format. The measurements will then also
be saved in the image file. They can at any time, be edited deleted or
augmented.
39
Measuring images
Outputting various measurement parameters
Task
Measuring areas
You want to measure the area of a workpiece. Have a variety of measurement
parameters, such as the area, the perimeter and the diameter, output. Have the
diameter shown in the image.
1.
Acquire or load an image.
2.
In the Measurement tool window, click the Closed Polygon button.
3.
Left click to define individual points on the edge of the object. Right click
to finish the measurement.
4.
Take a look at the result in the Measurement tool window.
•
Viewing the list of
measurement
parameters
5.
Outputting additional
measurement
parameters
6.
The illustration shows the image with a measurement object.
In the Measurement tool window, click the Select Measurements button.
•
In the dialog box you'll see a list with all of the available
measurement parameters. At the bottom of the dialog box you'll see
a list of the measurement parameters that are currently calculated for
all objects.
Go to the list of all of the available parameters, then click the Diameter
measurement parameter.
•
On the right, an illustration shows you how the parameter is
calculated.
You can see that there are different ways in which the diameter of a
2D object can be calculated.
7.
Click the Max entry in the list under the illustration, to select the Max
(Diameter) measurement parameter. This determines the workpiece's
largest span.
8.
Click the Add 'Max (Diameter)' button.
40
Measuring images
•
9.
Outputting
measurement
parameters in the
image
This measurement parameter will be added to the list of
measurement parameters to be calculated. All of these measurement
parameters will be displayed in the tool window.
Close the dialog box with OK.
10.
Take a look at the result for the diameter in the Measurement tool
window.
11.
Use the Tools > Options... command and select the Measurement >
Measurement Display entry in the tree view.
12.
Select the Text > Show value check box and close the dialog box with
OK.
•
Now the measurement values will be displayed in the image.
13.
Open the Select Measurements dialog box.
14.
At the bottom of the list of all of the calculated measurement parameters,
click the Max. (Diameter) measurement parameter.
15.
To the right of this list you'll see a button with a blue arrow. Click this
button to move the measurement parameter to the top of the list.
16.
Close the dialog box with OK.
17.
Take a look at the result for the diameter in the image.
Note: The measurement display in the image has to be updated before the new
settings are taken into account. You can update the measurement display by
adding another measurement or by selecting an existing measurement in the
image.
41
Measuring images
Measuring several images
Task
You want to measure cells on multiple images. To do so, acquire some images
and measure them one after another. Have the results from all images displayed
simultaneously. Take a look at the mean value for all of the measurements.
Loading images
1.
Acquire or load some images.
Measuring cells
2.
Activate the first image in the document group.
3.
Double click the Arbitrary Line button, located on the toolbar at the top of
the Measurement tool window.
•
4.
Measure the diameter of several cells.
5.
Activate the next image. Measure the diameter of several cells on this
image, too.
6.
Click the Arbitrary Line button again, and switch off the length
measurement.
•
Displaying the
measurement results of
all of the images
You will then switch to continuous measurement mode. In the
continuous measurement mode, you can immediately make further
measurements.
Cells have been measured on both images.
7.
Use the Tools > Options... command and select the Measurement >
Results entry in the tree view.
8.
Clear the Show measurement objects > Only of the active image check
box.
9.
Close the dialog box with OK.
•
Now the results for both images will be shown simultaneously in the
tool window.
•
Use the Document measurement parameter to display the name of
the image with which the measurement results are associated in the
results sheet. Now you can match the measurement results
unambiguously to an image, even if all measurement results are
displayed together in the tool window.
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42
Configuring the system
6.
Configuring the system
6.1. Overview
Prerequisite: Your software has been installed and the camera is connected to
the computer. The camera driver is installed in MS-Windows. When starting, your
software now recognizes the camera that is connected.
Process flow of the configuration
To set up your system, the following steps are necessary:
Configure the hardware
Calibrate the system
Configure the hardware
Every microscope is equipped with a different objective. In addition, your
microscope can also have a magnification changer or a camera adapter with a
particular magnification. Use the Acquire > Devices > Device Settings dialog box
to enter your microscope components.
You can also find all of the camera settings in the Device Settings dialog box.
Calibrating the system
When you have entered the objectives being used and, if necessary, the camera
adapter's and the magnification changer's magnifications, all of the images that
you acquire with your software are automatically X/Y-calibrated. Use the Manual
Magnification Calibration calibration process if this calibration is not precise
enough.
About the system configuration
When do you have to
configure the system?
You should configure and calibrate your system when you start the computer
after installing the software for the first time. If you later change the way your
microscope is equipped, you will need to change the configuration of certain
hardware components, and possibly also recalibrate them.
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43
Configuring the system
6.2. Configuring the system
Your software requires information about your camera, the objectives, and the
magnification of the microscope's camera adapter in order to be able to acquire
correctly calibrated images. That's why you need to configure the system.
Preconditions
Your software has been installed and the camera is connected to the computer.
The camera driver is installed in MS-Windows.
Configuring the specified hardware
1.
Use the Acquire > Devices > Device Settings... command.
•
Configuring the camera
Saving the hardware
configuration
2.
Select the Lightpath entry in the Sort by list.
3.
In the tree view on the left-hand side, expand the Camera > <camera
name> entry (Basler USB3 Vision for example).
4.
Select the Camera Adapter entry.
5.
Select your camera adapter's magnification in the Magnification list on
the right. The magnification is imprinted on your camera adapter. Typical
values would be 1.00 or 0.63.
6.
For your software, a typical hardware configuration is predefined. The
name of this hardware configuration is BaslerDefault. In the
Configuration group, click the Copy Device Configuration button.
7.
Enter a name for the new hardware configuration in the Copy Device
Configuration dialog box.
•
Configuring the
objective nosepiece
8.
10.
The copied hardware configuration is added to the list of the
hardware configurations.
Select the General > Manual Nosepiece entry in the tree view.
•
9.
The current configuration of the nosepiece is shown on the right side
of the dialog box. When you configure the software for the first time,
several typical objectives are preset.
Check the preset objectives and adjust the values if necessary.
Select the objectives with which your nosepiece is currently equipped
from the Magnification lists on the right. Start with the smallest
magnification, then a higher magnification, and so on. The magnification
is printed on the objective itself.
•
Configuring the
zoom/magnification
changer
In the tree view on the left side, you can find the hardware
components that are required for the correct calibration of the images
that are acquired.
In the Description field, a description of the objective will be
suggested. You may change the description of the objective in the
Description field, if you wish.
11.
If the objectives don't use air as their refraction medium, select the
immersion medium from the Refraction Index list. In this case, you find
an appropriate label on the objective.
12.
Select the General > Manual Zoom / Magnification Changer entry in the
tree view.
13.
Enter one of your magnification changer's magnifications in the
Magnification field and click the Add button.
•
All of the magnifications that have been entered will be available on
the Microscope Control toolbar.
44
Configuring the system
•
Finishing the
configuration of the
system
14.
Close the Device Settings dialog box with OK.
•
15.
If your microscope doesn't have a magnification changer, make the
default settings yourself.
You may get a message asking you to check the calibrations. You
can perform the calibration either now or later.
Use the View > Toolbars > Microscope Control command to display the
toolbar.
•
The Microscope Control toolbar contains buttons with all of your
objectives, correctly color coded.
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6.3. Manual Magnification Calibration
All images you acquire using your software are automatically X/Y-calibrated. Use
the Manual Magnification Calibration calibration process if this calibration is not
precise enough. You need a calibration standard to perform the calibration
process.
How is the preset
magnification
calibration determined?
The X/Y-calibration of an image you have acquired with your software is
calculated from the size of a pixel and from the total magnification. The total
magnification at the time of acquisition is usually a combination of the objective
magnification and the camera adapter's magnification. If you are using a
magnification changer, this will also have an effect on the total magnification.
The preset magnification calibration uses the hardware components from the
Device Settings dialog box and the camera's pixel size, which can be read out
from the camera driver.
Starting the manual
magnification
calibration
Returning to the preset
magnification
calibration
Use the Acquire > Calibrations... command. Select the Manual Magnification
Calibration entry in the Calibrations dialog box. Then click the Calibrate... button.
When you have manually calibrated your objectives, you can at any time return
to the preset values. Select the Manual Magnification Calibration calibration
process in the Calibrations dialog box. Then click the Edit... button. Clear the Use
manual calibration check box in the Manual Objective Calibration dialog box.
45
Configuring the system
Calibrating the objective manually
1.
Lay a calibration standard under your light microscope.
2.
If you are using a magnification changer, set the magnification changer
to a magnification of 1 and select the magnification 1x on the Microscope
Control toolbar.
3.
Start the Manual Magnification Calibration calibration process.
•
In the Calibrate objective list all of the objectives that you have
registered with your software are listed.
•
All of the objectives that have already once been manually calibrated
.
are identified by a check-mark
4.
Select the check box in front of each objective that you want to calibrate.
5.
Click the Next > button.
•
Your software will automatically switch to the live mode.
6.
Bring the objective you want to calibrate into the light path. The dialog
box shows the name of the objective.
7.
Focus on the calibration standard.
8.
Click the Next > button.
9.
Click the Set Reference Distance button.
•
The image is acquired and live mode is closed.
•
The mouse pointer appears in the image window.
10.
Determine the starting and end point of the calibration reference distance
with your mouse pointer, for example, a distance of 800 µm. You define
these two points by clicking with your left mouse button.
11.
Click the right mouse button and select the Confirm Input command in
the context menu.
46
Configuring the system
•
12.
The Set Reference Distance dialog box opens.
First select an appropriate unit
.
If, for example, you are using a calibration standard with a length of 1
mm and 100 scale units, select the unit µm.
•
Note: If you select the unit in here, this image will be calibrated in
inches. That doesn't mean, though, that all of the images will now be
acquired with in as the basic unit. You specify the basic unit for the
image acquisition in the camera settings, in the Acquisition Settings
> Acquisition > General dialog box.
13.
In the Length field, you enter the length of the calibration standard that
you want to use for the calibration. With a calibration standard that is 1
mm long for example, you can enter 800 in this field if you have defined
the reference distance between the scale divisions 10 and 90.
14.
Click the OK button to confirm the calibration.
15.
Click the Next > button if you want to calibrate further objectives, or the
Finish button when you don't have any more objectives to calibrate.
16.
Repeat the manual calibration for the other objectives that you've
chosen.
•
17.
Check the results and close the Manual Magnification Calibration dialog
box with OK.
•
18.
In the Calibrations dialog box, the Manual Magnification Calibration
calibration process will then have the Calibrated status.
Close the Calibrations dialog box.
•
19.
When the last magnification calibration has been carried out, the
Manual Magnification Calibration dialog box opens.
When you then acquire an image, the image's X/Y-calibration will be
calculated with the new magnification values. Please note that before
you make the image acquisition, you will have to select the objective
you want to use, in the Microscope Control toolbar. If you are using a
magnification changer, you will also have to select the magnification
level used.
You can view the actual objective magnifications resulting from the
manual magnification calibration at any time.
Select the Manual Magnification Calibration calibration process in the
Calibrations dialog box. Then click the Edit... button.
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