SPOT 4.1 Basic and Advanced Software Manual for SPOT Insight ...

Ch. 17 – Using Other Mac Imaging Application in Basic ModeImage TypeBrightfieldtransmittedlightDarkfieldtransmittedlightFluorescenceNomarskiDIC/HoffmanPhase contrastPolarized lightBrightfieldreflectedlightDarkfieldreflectedlightStereo/Macro (allthree options)White Balance TechniquePosition the slide so that the specimen is not in the field of view, but light is going through the slidenear the specimen.Perform the white balance using the brightfield mode on your microscope. See Brightfield–transmitted light above.Use the Match Color feature (see above)For Nomarski DIC: Adjust the Nomarski prism until the image is in either the “brightfield” or the“gray scale” view.For Hoffman: Perform the white balance directly on the Hoffman image.Position the slide so that the specimen is not in the field of view, but light is going through the slidenear the specimen.Pull all polarizers and compensators out of the light path and then position the slide so that thespecimen is not in the field of view, but light is going through the slide near the specimen.Replace your sample with a mirror.Perform the white balance using the brightfield mode on your microscope. See Brightfield –reflected light above.Using an EPI-illuminator (shining a light on the top of your specimen from off to one side), replaceyour sample with a white sheet of paper.5. Click on Begin to start the calculation. The camera computes the white balance values, and they areautomatically saved to the image settings for the image type that you selected.Match ColorThe Match Color feature is typically used in two situations:• You want to change the colors of an image on the screen to match the way that you think the specimenlooks.• You want to set the white balance for pictures taken with a microscopic technique that never has whitein its images. A prime example of this situation is fluorescence microscopy.To use the Match Color feature, follow these steps:1. Click on the Preview button to capture an image. The Match Color option remains inactive until apreview image is captured.2. Select Match Color. The cursor becomes an eyedropper.3. Using the mouse to move the eyedropper, position it on the pixel location of the color that you want tomatch. The R, G, and B values of the pixel that you select must all be less than 255.4. Click on the pixel. The Match Color dialog appears with information about the pixel that youselected:− Hue – The actual color, as distinguished from others in the color spectrum. Hue is derived froma color wheel and is expressed in the color's angular location (e.g., 0 to 360 degrees).− Saturation - The proportion of perceived pure hue in the color. Saturation (in the SPOTsoftware) is measured on a numerical scale of 0 to 100, where zero equals pure gray and 100equals pure hue (e.g., “day glow”).5. Move the cross hairs to the desired color either by clicking on the new location or clicking anddragging. As you move the cross hairs, the color in the image changes accordingly, allowing you topreview the color before you save the image. Alternatively, you can click on the Make White/Greybutton to automatically white balance the image.6. Click on Ok.User Guide to the SPOT Insight Camera 307