Using the Diagnotics Instruments Spot Camera

Using the Diagnotics Instruments Spot Camera
Using the Diagnostics Instruments Spot Camera
By Dr. Steve Spilatro
(Edited by Dr. Dave McShaffrey, who introduced all the errors)
The Spot camera is a cooled CCD digital camera of very high resolution, light sensitivity,
and cost -- a camera of this type costs around $7000 - $9000. Thus, care must be taken in
its use.
The camera is mounted on the microscope, as shown in the figure
to the right.
Turning on the camera
The camera must be turned on at the power supply
(shown to the right), located near the computer.
First turn on the power strip along side the computer,
then turn on the camera power supply. The red
indicator light will illuminate when the power supply is
Acquiring Images from the Spot camera
Open the Adobe PhotoShop program by double clicking on the PhotoShop
From the ‘File’ drop-down menu, choose the ‘Import’
Click on ‘Select Twain_32 Source’
In the option window that appears, select the ‘Spot
Camera’ as the import source.
Opening the Image Import Window
To open the Spot camera window, under ‘Import’
On the ‘File’ menu, select ‘Twain_32’
This will open the camera Image Import
Window. This is the window you will use to
control the camera.
Camera Controls
Make sure under Image Setup that
‘Brightfield’ (not #Brightfield) is
** Do not Add, Modify or Delete
the Image Setup setting ***
The Image controls that you may use
are indicated to the right.
‘A’ is the ‘Focus’ control
This is used to focus the image as
viewed through the camera.
‘B’ is the ‘Framing’ control this is used to
position the image in the frame as
observed through the camera.
‘C’ is the ‘Exposure’ control
This sets the proper exposure for
the camera. Normally you will not
use this since the exposure will
be set automatically when you
shot the image.
‘D’ is the ‘White balance’ control this is used to assure that the background color is white.
The ‘Preview’ button allows viewing the image before importing to PhotoShop.
The ‘Acquire’ Button shots the picture and returns it to PhotoShop.
Standard procedure for importing an
Turn on the microscope.
The power switch is a small toggle on the back, right-hand
1. Setting the illumination level
The illuminator control panel is on the lower right-hand side of
the microscope.
Move the slider level to approximately so that the lights in
positions ‘4’ or ‘6’ are illuminated.
If the slider level has no effect, push the ‘preset’ button.
It is best to set this once and then not to change it during a
photo session. If this is changed after the ‘white balance’ is
set (see below), then the white balance may need to be reset.
2. Setting the white balance.
Position the microscope slide to a clear area (no tissue) Click
the ‘white balance’ control button, and then click ‘Begin’
After the process is complete, click OK to save the white
balance settings to the ‘Brightfield’ setup.
If you get an “Exposure too short…” error message, see notes below about adjusting the
light level.
3. Adjusting the image brightness and contrast
Possibly the most important adjust you will make is to image
brightness and contrast.
These are normally adjusted using the substage diaphragm,
located under the microscope stage, as show to the right.
In general, the substage diaphragm must be nearly closed
under low magnification, and opened as the magnification is
increased. Opening the substage diaphragm increases the
brightness of the image but decreases the contrast, and these
two must be balanced to get the best possible image.
Too much light
The Spot camera is so sensitive that an error
message will appear if you try to set the focus,
exposure or white balance for an image that is too
When this error occurs, the brightness must be decreased. This can be done in several
ways, including:
1) closing the substage diaphragm
2) using the neutral density filters (see below)
3) using the illuminator preset button (see below)
However, it’s best not to adjust the microscope illuminator slider lever if the white balance is
already set.
Using Neutral Density Filters
Neutral density filters decrease brightness
without changing the ‘color’ of the light. The
‘ND6’ and ‘ND25’ filters are located below
the mechanical stage knobs on the righthand side of the microscope. Flipping the
levers adds or removes the filters from the
light path.
Using the Illuminator Preset button
The Illuminator Preset button is a convenient way to easily switch
between a brightness that is suitable for viewing an image through
the microscope, and a brightness that is low enough for the Spot
Push the button to increase the brightness when viewing the
specimen through the microscope, and push it again before taking
the picture.
Note: make sure the white balance is set for the image
under the low (preset off) level.
4. Setting the focus
Reposition the microscope slide so that specimen to be
photographed is centered.
Clicking the ‘Focus’ command button will open the
window shown to the right.
Click “Begin” to start focusing
The camera will start taking a series of rapid-fire, black
and white images. Depending upon the predominate
colors in your image, the filter color may be set to red,
green or blue (sometime this matters, sometimes not).
The image can be focused by turning the focusing knobs on the
microscope stage, or by turning a focusing ring located just above
the microscope on the camera mount, as shown to the right. The
advantage of using the camera focusing ring is that the image can
be in focus when viewed through the microscope and through the
NOTE: it is best to focus on part of the specimen that has high
contrast, since the focusing image very ‘grainy’. In general, the
image observed while focusing never look very sharp.
Click ‘Stop” when you are done focusing, and close the focus
5. Framing the image
The Framing control can be used to check and make
adjustments to the composition of the image.
Click the ‘Framing Control button to open the Frame Window,
as shown to the right.
Click ‘begin’ to …. begin.
It works much like the focus window; a series of quick images
are taken, so that you can adjust the position of the specimen
using the mechanical stage knobs on the microscope.
When the image suitable positioned, click ‘stop’ and close the window.
6. Taking the picture and passing it to PhotoShop
Click on the Preview button on the right to see what your image will look like. If it looks
good, you can just click on the Acquire button to move it into PhotoShop.
Close the Image Control Window; the image is now ready for work in PhotoShop.
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