Using the AmScope Microscope Cameras

Using the AmScope Microscope Cameras
Using the AmScope Microscope Cameras
Part 1 – Setup.
In order to use the camera, you will need: a)
the camera system; b) a computer running the
camera software.
The camera system is contained in a Pelican
Case (see image to right). In the case there
should be the camera, the camera lens cover,
two adapter rings, and a USB cable.
The camera is designed to replace the ocular
lens (see image below). It can also be placed
in a trinocular tube, or can replace the eyepiece
in a dissecting scope or spotting scope.
Steps in setting up the camera.
1. Install software. The software, called
ToupView, is installed on several of the
department’s laptops. If it is not installed on the
computer you are using, install it. The software is located on the intranet, in the
Biology/General/Programs/Toupe directory. Run the autorun.exe file, and install the Camera
Drivers and ToupView software.
2. Set up the microscope. Set up the microscope as you normally would. Pay particular attention to
making sure all parts of the scope are clean of dust, oil, fingerprints, etc. If you have not been trained
(microbiology, cell biology) in routine cleaning of the microscope, get help from a faculty member if
the scope is dirty. Carefully remove the ocular from the microscope and place it in the Pelican case
where the tube of the camera was. Remove the cap from the end of the camera, and place the
camera into the eyepiece tube of the microscope. Do this quickly, so there is no opportunity for dust
to get into the microscope or into the camera. Place the camera end cap in the Pelican case so it
doesn’t get lost. The camera tube should move freely in the microscope ocular tube – if it rattles or
there is excessive play (i.e. the camera tube is way too small) add one or the other of the adapter
rings from the Pelican case.
Using the AmScope Microscope Cameras.docx
1
3/7/2012 10:07:00 AM
3. Connect the camera and start the software. Connect the camera to the USB port of the computer
using the USB cable from the Pelican Case. This should cause the computer to begin installing the
driver for the camera (unless the software had already been installed and the camera used with the
computer in the past). Locate the ToupView software on the computer (usually an icon on the
desktop) and start it.
ToupView software and important icons:
1.
2.
3.
4.
Live Capture – selects, starts camera
Video source properties – adjust most exposure settings here.
Video stream format – adjust video image size here.
Capture still image – take still images here. Note: Do NOT use the camera icon; this
takes lower-resolution views.
5. ROI – allows you to select a different Region of Interest
6. IF – manual fusion – allows you to stack focus several images.
7. Live Capture window – shows a live view from the camera.
Using the AmScope Microscope Cameras.docx
2
3/7/2012 10:07:00 AM
Setting up the slide:
1. Put a slide on the stage and set the scope to the lowest possible magnification.
2. In the ToupView software, click on the Live Capture Icon and select the camera (it should be
the only camera there).
3. The Live Capture window will now open.
4. Adjust light levels – set the microscope rheostat (light source) to 5 or 6 and close the iris
diaphragm fully. The camera should automatically adjust.
a. If there is a pattern of horizontal or vertical bars you are getting too much light, and the
camera is taking the image so quickly that it is capturing the flickering of the light
source. Reduce the light, either with the rheostat or the iris diaphragm.
b. If you have too little light you will get noise 
5. Focus the scope and scan for an area of interest. Center the subject and move to a higher
power if needed. Readjust light if needed.
Adjusting the software:
1. Be sure the video size is maximized (2592 x 1944) using the Video Stream Format icon (#3,
above)
2. Open the video source properties using the icon (#2, above).
3. On the ROI tab (below) move the ROI to a blank area of the slide.
Using the AmScope Microscope Cameras.docx
3
3/7/2012 10:07:00 AM
4. On the color tab, click the “One Push” button to
get an approximate color temperature setting.
5. Fine tune the color setting by sliding the
Hue/Saturation and Brightness controls (if
needed)
6. Normally, you should not adjust the contrast or
Gamma controls.
7. Move to the exposure tab.
a. Adjust the Target slider to get a good
overall exposure. Don’t worry too much
about pure whites or blacks at this point.
b. Note that as you adjust the target slider,
the exposure time is adjusted. If it gets
too big (above 100ms or so) then the
camera delay will be very long and the
camera will be slow to track moving
objects (or allow you to follow the slide as
you move it). If this is the case, add light
via either the microscope rheostat or the
iris diaphragm, being careful not to get any
horizontal or vertical bars. Readjust the
target slider.
c. In extreme cases you may need to
unclick Auto Exposure and set the
exposure time manually.
8. Go to the Histogram tab. If you need to get
pure whites or blacks you can adjust the
histogram via the boxes below the histograms or
by dragging the pink lines. Watch the image as
you adjust the histogram.
Using the AmScope Microscope Cameras.docx
4
3/7/2012 10:07:00 AM
9. On the Misc. tab you can make several other adjustments:
a. If the video does not run or is choppy,
you can adjust the frame speed level.
On slower computers you may need to
slide it to the right. If the computer is
really slow, you may need to decrease
the size of the video (use the Video
Stream Format icon).
b. You can try adjusting the Light
Frequency if you are getting noticeable
flicker. In theory, you should set it to
60 Hz.
c. If you can’t read a stage micrometer,
try clicking one or both of the flip
buttons. This might also help you
move the slide in a predictable fashion.
Take the picture:
1. Use the Capture Still Image (S), NOT the Capture Image (camera) icon.
2. Save the image (File:Save As), normally as a bmp or tiff file to avoid compression.
3. See the scale bar procedure below if your image will require a scale bar.
Take a video:
1. Use the menu item Capture:Start Video Capture
2. You will have to answer several questions in a dialog, including the file name – this may take
several seconds to do.
3. When you complete the dialog, video capture will start, and progress can be monitored on the
status bar at the bottom of the window.
4. When you have captured what you want, use the Capture: Stop Video capture on the menu
(you can also set a time limit in the dialog; if you do so the video will automatically stop at that
point).
Notes: I have not tested the various video settings, so I can give you no advice at this point. Also, in
some cases you might want to use the time-lapse icon to capture frames less frequently than the
video does. This will still give the appearance of motion for things that take minutes to noticeably
change. The camera will take a series of still images which can later be combined into a video.
Using the AmScope Microscope Cameras.docx
5
3/7/2012 10:07:00 AM
Image Stacking:
You can take a number of images at different focal points and stitch them together using the
ToupView software or Photoshop. This allows you to get much greater depth of field.
ToupView Technique:
1. Set up the slide as you normally
would.
2. For capture, use the manual
fusion icon (IF).
3. This brings up a dialog. Focus to
the top of the object you are
interested in, and click the
capture button on the dialog.
4. Focus down slightly with the fine
focus knob and capture another
image
5. Continue this process until you
have focused all the way through
the object – this may be 3-10 or
more images.
6. Click on the Fusion button. The
software will take 10 seconds to
several minutes to work its way
through the images, combining the
in-focus parts of each.
7. Save the file and add a scale bar
as you normally would. You
should add a letter to the file
name, text to the image, or data to
the file’s metadata to indicate that
it is a fused image.
Using the AmScope Microscope Cameras.docx
6
3/7/2012 10:07:00 AM
Photoshop Technique:
1. Take multiple images, starting from the top down with slight focus changes each time (as done
in the ToupView technique, but manually using the capture still image icon then saving the
files).
2. Initial Processing
a. Close all programs except for Bridge!!!!!
b. Open the folder you saved the ToupView images to in Bridge
c. Using Bridge, select all the photos you want to stack
d. Use the Tools:Photoshop:Photomerge menu item
e. Unclick “blend images together” and click OK
f. When file is completely open in Photoshop close Bridge
3. Final Processing
a. In Photoshop, crop and resize image. Make image as small as possible for your needs
to avoid memory problems and to decrease processing times.
b. Select all layers
c. Use the Edit:Autoblend Layers command and blend images together option
d. Save the resulting file as a PSD file
e. Use the Layers:Flatten Image menu item to flatten the image
f. Sharpen and save as a JPG or other file.
Notes: At present I cannot recommend one technique over the other. At first glance, using the
ToupView technique is much quicker, but the image quality does not appear to be as good.
Using the AmScope Microscope Cameras.docx
7
3/7/2012 10:07:00 AM
Below: Image stacked with Photoshop. The algal filament to the right moved between photos and the
algorithm for stacking ghosted the image with a lot of color. On the other hand, image detail is good
in the areas where there was no movement.
Below: Image stacked with ToupView. This image is pretty good overall, and has almost as much
detail as the Photoshop stack. However, it is harder to control details of exposure in this case.
Notice that the ToupView handled the ghosting of the moving filament better (although to be fair, it
didn’t move as much).
Using the AmScope Microscope Cameras.docx
8
3/7/2012 10:07:00 AM
Scale bar procedure:
If you want a scale bar on your micrograph (you should) you need to photograph something of known
scale. In most cases, this will be a stage micrometer, a microscope slide with a tiny scale, usually
1mm long graduated in 0.01mm units. On a dissecting scope, you could photograph a normal
millimeter scale.
1. It is essential that you photograph the scale at the same magnification and quality (pixels) that
your micrograph is photographed at. Be sure to use the same capture button and record the
objective in use. If using a dissecting scope, DO NOT ADJUST THE ZOOM.
2. Photograph the scale at the same magnification as your micrograph. Save the file, ideally
with a file name that includes the magnification (scale_100x.bmp).
3.
4.
5.
6.
7.
8.
Open the scale bar file and use the Select:All command to select the entire image
Use the Edit:Copy menu command to copy the scale bar to the clipboard
Open your micrograph in Photoshop.
Add a new layer (call it scale bar image)
Use the Edit:Paste command to paste the scale bar from the clipboard to the new layer.
If needed, use the Edit:Transform:Rotate command to rotate the scale bar until it is straight –
do not worry about any background that shows, or parts of the scale bar that get clipped.
9. Add a third layer (name it scale bar)
10. Use the line tool, set to a reasonable thickness (weight)
and with a color that will contrast with the background of
your micrograph. Holding down the shift key (to get a
straight, horizontal line) draw a line over a known,
representative area of the scale (for instance, 0.1 mm of the
one mm scale at 100x) (see image to right)
11. Hide the scale bar image by unclicking the eye icon to the
left of the scale bar image layer on the layers palette.
12. Use the moving tool (at the top of the toolbar) to drag the drawn scale bar to a blank area on
the micrograph (usually the lower right corner).
Using the AmScope Microscope Cameras.docx
9
3/7/2012 10:07:00 AM
13. Use the Type tool to type in the length of the drawn scale bar. Remember to place a 0 in front
of any decimal point (i.e. 0.1mm instead of .1mm). The thickness of the type should be
approximately the thickness of the scale bar.
14. If the scale bar and/or legend doesn’t stand out from the background, you may want to doubleclick on the layer and adjust the layer styles, Adding a contrasting stroke color is often an
effective tactic. Be sure to treat both the scale bar and the text to the same style.
Using the AmScope Microscope Cameras.docx
10
3/7/2012 10:07:00 AM
Alternate scale bar technique:
1. Using the Analysis:Set
Measurement Scale:Custom
dialgog, draw a line over the
image of the scale and set the
scale. Click OK.
2. Use the Analysis: Place Scale Marker menu item to
bring up the next dialog. Set the length as needed
and set the font size. Note: the bigger the image is,
the bigger the font size will need to be. It is not
unusual to use a size 48 or bigger font).
3. Hide the scale bar image layer by unclicking its eye and thus showing the new drawn scale
bar on the micrograph instead.
4. After the scale bar is placed, use the move tool to relocate it as needed.
Using the AmScope Microscope Cameras.docx
11
3/7/2012 10:07:00 AM
Putting things away:
1. Computer:
a. Close the ToupView Software.
b. If you are done with the computer, shut it down.
c. Neatly coil the power cords.
d. Return the computer to its designated place.
2. Camera:
a. Unplug the camera from the cord, and unplug the USB cord from the computer.
b. Remove the camera from the microscope and immediately replace it with the eyepiece.
c. Immediately cap the camera and place it in the Pelican Case.
d. Replace any spacer tubes used in the Pelican Case.
e. Close and lock the Pelican Case
f. Carefully coil the USB cable and place it in the Pelican Case
g. Return the case to its designated place.
3. Slides/specimens:
a. Remove any slides from the microscope. Slides should be cleaned for re-use.
Coverslips can be discarded in a SHARPS BOX ONLY.
b. Be sure to replace any stage micrometers where they were obtained.
c. Return any specimens to their designated place in the same condition you got them (in
general, lids should NOT be screwed on tight).
4. The microscope should:
a. Have the lowest power objective (or blank) in place over the stage
b. Have the stage raised to its highest point
c. Have the eyepiece directed back over the body of the scope (pointed towards the focus
knobs)
d. Have the power switch turned off
e. Have the cover in place
f. Have the power cord coiled neatly around the base
g. Be replaced where it belongs.
5. Work area:
a. The work area should be cleaned up and wiped down with disinfectant.
b. All materials should be returned to their designated places.
Using the AmScope Microscope Cameras.docx
12
3/7/2012 10:07:00 AM
Was this manual useful for you? yes no
Thank you for your participation!

* Your assessment is very important for improving the work of artificial intelligence, which forms the content of this project

Download PDF

advertising