Measure fluorescence ratio with magic wand-based ROI drawing

Parameters:

Name	#	Type	Description
Channel Number (Numerator)	1	integer	Channel number associated with the image. Use Edit/Rename to view or change channel number.
Channel Number (Denominator)	2	integer	Channel number associated with the image. Use Edit/Rename to view or change channel number.
Background Level (Percentile)	3	real	Background is calculated as frame-by-frame mean of those pixels that are the darker than this percentile of image histogram for the duration of the entire recording.
Cell diameter (pixels)	4	integer	Cells ranging around this size will be selected

Description:

This pipeline computes ratio image and shows it in Intensity Gated Pseudocolor, draws ROIs and plots ratio values as ratio of the average fluorescence intensities measured in the two source images for each individual cell. The fluorescence ratio is calculated as the ratio of mean fluorescence intensities allowing working at low signal to noise ratio. This pipeline is a simplified workflow. The magic wand-based automatic ROI drawing is used here.
This pipeline is applicable to Ca2+-imaging (e.g. Fura-2, yellow cameleons), other ratiometric ion probes imaging (e.g. PBFI, SPFI, SNARF-1, BCECF), reactive oxygen species imaging (e.g. HyPer, roGFP), and intramolecular FRET sensor imaging.

Version history:
V2
Changed caption