Fluorescent Double-Labeling

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Objective 5x Zoom 4.0 x (except PBN as 3.0x) Format 1280x1024

Live (play button icon)

Adjust AutoExposure/Analog Gain or enter set exposure time

Acquire -> Capture (camera icon) captures image with flurophore channel (e.g. "FITC") (have to restart Live after a capture to view live image again)


Save As -> Tagged Image Format (TIFF) proprietary Nikon file format that preserves acquisition parameters

Using TIFF files, Drag and drop one channel from one TIFF file onto second channel in second TIFF file to create a merged TIFF file. e.g., drag FITC on top of TRITC channel

Save merged image as TIFF file

Save As JPEG Save merged image as JPEG file to convert from emission filter channels to RGB

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