STARTING PARAMETERS for capturing images with a ODCM camera

• Open ToupView™ (ScopePhoto™ ScopeTek™) and focus the specimen in the live capture window.
• Select Video Source Property from the Setup menu.
• White Balancetab
  • Hold a sheet of white paper under the microscope so that the light source is projected onto it. Click Auto White Balance. The colour temperature will be set to a neutral grey (~20%).
• Exposuretab
  • Tick Auto Exposure and click the Default button.
• Colourtab
  • Adjust the gamma, saturation, and contrast as desired.

Note: The white balance will not change unless the colour temperature is changed. This can occur when increasing the intensity of a light source, changing the bulb, or using a different microscope. Using the white paper method to achieve a neutral grey will adjust the white balance to different light sources/intensities.

Information:

http://en.wikipedia.org/wiki/Contrast_(vision)
http://en.wikipedia.org/wiki/Exposure_(photography)
http://en.wikipedia.org/wiki/Image_editing
http://en.wikipedia.org/wiki/Dynamic_range
http://en.wikipedia.org/wiki/Gamma_correction

CALIBRATION

Required is a known length for calibration (e.g. a stage‐micrometer). Place this under the microscope so that it is visible in the computer's monitor.

1. Ensure the video window is set to 100%.
2. Select the "Define Software Power" icon. Choices are: horizontal line, vertical line and any line. For calibration purposes, horizontal line is recommended.
3. A cross‐line will be super imposed. Using the mouse drag the cross‐line to a starting point, then again, left click, and then move to the measurement's end‐point and click again. Consider. Measurements may be more accurate if always starting and end‐points always are from the beginning of a graduation. The calibration should include as much of the calibration ruler as can fit onto the screen at the used magnification setting. The centre of the screen will be the least distorted part of the image.
   
4. When the end‐point of the measurement has been selected, the "Define Software Power" window appears. Index 1 and 2 and co‐ordinates denote the location on the image where the measurement was made based on the horizontal and vertical rulers surrounding the image.
5. Under "Actual Length" enter the length that was measured and select the unit of measurement. Then select "OK".
6. The "Software Power" window will appear. Select "Add".
7. Enter the total magnification (i.e. objective x eyepiece power) then click "OK". This will be used indicative, to identify a particular calibration, but is unlikely to be accurate.)
   This completes the calibration for this particular magnification. Click "OK".
   Repeat steps 1‐7 to calibrate other magnifications on your microscope. All calibrations will be stored in the system for future measurements.

Note: If other microscopes are being used with the software, they will also have to be calibrated.

MEASURING

Before measuring, ensure draw preferences are set as desired. Select Options and then Draw.

Note: Ensure correct "software power" is set (the nominal magnification on the microscope)

Measuring on a live image (e.g. not a jpg)

1. Quick measurements can be made on the live image. Click the ruler icon and select the type of measurement required (length or angle).
2. A crosshair will appear allowing point‐to‐point length or angle measurements to be made (depending on the type of measurement selected).

Note: These one‐off measurements will not be saved.

1. Batch measurements can be made for angle and length measurements. Select Batch Measurement and the type of measurement required. 
2. Again, a crossline will appear. Make the measurement as described above. The batch measurement function is particularly useful for making the same measurement on multiple samples. 
3. When measurements are complete, click "View Results".
   
4. The following window will be displayed detailing all measurements made.
5. Click the "Process" button to export results to Excel or a text file. Batch results may also be averaged. Batch measurements will only be saved if exported to Excel or a text file. Closing ToupView will delete all batch measurements. Before starting a new batch of measurements delete any other.

Measuring on a captured image

1. Measuring on a captured image has two advantages. Measurements can be saved on the image (useful for reporting/archiving) and they can be exported to Excel (image and measurement data).
2. A measurement made on a captured image is stored on a layer over the raw image.
3. After captured the image, select Layer from the menu and then New.
4. Enter a name for this new layer.
5. Inserting a layer on a raw image will activate the "Draw" menu and icons. Select the measurement type required. Select start and end point of measurement.
6. Further measurements can be made on the same layer or new layers can be inserted for different measurements. Multiple layers are useful if a number of measurements need to be made on the image. Layers can be made visible or hidden as required.
   
7. To save the image (with layers), select Save As from the File menu.
   
8. For a raw image and layers to be saved as one file, it must be saved as the ToupView file type (.sft). This will allow the image to be opened in ToupView at a future date and for layers to be manipulated or added to ‐ if necessary. Saving as any other file type at this stage will not save layers.
   
9. To print an image select Print from the File menu.
   
10. The saved image can also be exported to Excel. The image including data will be exported to allow insertion in spreadsheets as required.
    
11. If Export to Image is selected, a new image file will appear on the screen. Use the Save As function to save the file (complete with layers) as another file type (e.g. .bmp, .jpg etc.) for easy insertion in other programs, reports etc.

COUNTING

1. Counting how many particles with an image: With the desired captured image in focus, select "Plugin" from the "Menu Bar" and click "Count"
   
2. To set the parameters within this plugin, click "Setting" → "Parameter"
   
3. This panel will allow you to set your desired settings.
   
4. Once you have adjusted your parameters, press "M&C" → "Count" The program will allocate a set number to each particle the is found
   
5. Clicking on "Show All Results In Image"
6. Selecting "M&C" → "Check the Result" display a list of found particles shown below.