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

Ch. 4 – Advanced Program ModeTo draw the line, follow these steps:a. Position the crosshair cursor over the stage micrometer mark that represents the start point of yourcalibration line. In the illustration below, this is the 20 mm mark.b. Left click on the mouse.c. Position the crosshair cursor at the stage micrometer mark that represents the end point of yourcalibration line. In the illustration below, this is the 23 mm mark.d. Left click on the mouse again. The length of the line in pixels appears on the Calibration Setupwindow. To re-draw a line, double click anywhere on the image. This clears the previouslydrawn calibration line from the screen.9. Enter the length and units of the line that you drew (e.g., 3 mm).10. Click on OK. The 4x objective is now calibrated to perform measurements. Repeat steps one throughten for each objective.PalettesThe Palettes option on the Setup menu allows you to define new palettes and edit existing palettes. SPOTcolor palettes allow you to apply specific hue and saturation values to monochrome (8 bit or 12 bit) images.Palettes are useful in fluorescence applications where a sample is fluorescing at a single wavelength, andyou want the acquired image to have the correct color. For such a case, you could create a palette torepresent the color of the fluorescent dye and then apply that palette to the captured image. There are twoways to apply a palette to an image:• You can associate a palette with an Image Setup, as discussed in Ch. 5 - Preparing to Take a Picture.• You can apply a palette to a captured image by selecting the Set Palette optionfrom the Edit menu. Refer to Ch. 8 – Viewing, Editing, and AnnotatingImages for more details.To define a new palette, follow these steps:1. From the Setup menu, select Palettes.2. Click on Add. The Color Palette dialogappears as shown here:3. Either use the Wavelength or Hue andSaturation selection lists or move the crosshairson the color wheel to enter the values−−−Wavelength allows the user to specifythe exact wavelength ranging in valuefrom 400nm to 700 nm. Wavelengthswill automatically correspond to variousHues at full (100%) saturation.Hue is the designation for the actual color, as distinguished from others in the color spectrum.Hue is derived from the color wheel and is expressed in the color's angular location (in degrees) onthe wheel. (e.g., 0° to 360°).Saturation is the proportion of perceived pure hue in the color. Saturation (in the SPOT software)is measured on a numerical scale of 0 to 100, where zero is pure gray and 100 is pure hue (e.g.,day glow).4. Click on OK.User Guide to the SPOT Insight Camera 71