Fluorescent Microscope Image Acquisition

Startup Program: Open Spot Advanced program on desktop

Live image viewing: Click on Live button on far right of main screen. Controls button allows adjustments of gamma (grayscale range), contrast and brightness (essentially affecting exposure time). Snap button captures the current image.

New exposure setup: Setupà Image Setups à Add à (any)

Change exposure time to manual.

Then click on manual on left side, and set an exposure time.

Most of the other settings remain the same for all.

Color: Edit à Set Palette à (Choose palette to match stain)

Setup à Palettes… (Allows creation of new palette)

Adjust Contrast: Edit à Adjust RGB à Adjust Histogram (this is the best option)

Edit à Adjust RGB à Contrast (also may be helpful)

Edit à Adjust RGB à Adjust Gamma (adjusts grayscale range)

Multiple Stains: Create separate images using the different filters.

Then merge the images: Edit à Merge Images

Can also merge light images with fluorescent images

Sharpen Image: Edit à Filter à Unsharp image

Be careful, only so much is reasonable to believe.

Add Scale Bar: Edit à Set Calibration