Line 10:
Line 10:
*Acquire -> Capture (camera icon)
*Acquire -> Capture (camera icon)
:captures image with flurophore channel (e.g. "FITC")
:captures image with flurophore channel (e.g. "FITC")
−
:(have to restart Live after a capture to view live image again)
+
:(have to restart Live after a capture to view live image again
−
*Save As -> Tagged Image Format (TIFF)
*Save As -> Tagged Image Format (TIFF)
:proprietary Nikon file format that preserves acquisition parameters
:proprietary Nikon file format that preserves acquisition parameters
+
+
*Save As -> JPEG
+
:Save single fluorophore channel image as JPEG file to convert from emission filter channels to RGB
+
+
JPEG file of one fluorophore channel will have data in 2 or 3 RGB channels, so may need to delete unwanted RGB chanels (e.g. delete red channel from FITC JPEG, to leave only green channel.) Or, better still, convert fluorophore image to monochrome (i.e., so that luminance of flurophore channel is encoded as equivalent gray value from 0-255 in each RGB channel of the JPEG).
+
+
*Open JPEG files for each fluorophore/
+
:(e.g., FITC JPEG and TRITC JPEG)
*Using JPEG files, Drag and drop one channel from one JPEG file onto second channel in second JPEG file to create a merged JPEG file.
*Using JPEG files, Drag and drop one channel from one JPEG file onto second channel in second JPEG file to create a merged JPEG file.
Line 20:
Line 27:
*Save merged image as JPEG file
*Save merged image as JPEG file
−
−
*Save As JPEG
−
:Save merged image as JPEG file to convert from emission filter channels to RGB
−
−
JPEG file of one fluorophore channel will have data in 2 or 3 RGB channels, so may need to delete unwanted RGB chanels (e.g. delete red channel from FITC JPEG, to leave only green channel.) Or, better still, convert fluorophore image to monochrome (i.e., so that luminance of flurophore channel is encoded as equivalent gray value from 0-255 in each RGB channel of the JPEG).