Hi mr, In my experience with confocal & CCD work you cannot compensate data collected after acquisition, although saying that Molecular Dynamics used to sell Unix software called 'Imagespace' that apparently/theoretically could do compensation - never tried it myself as it seemed impossible to tell which part of the image data needed compensating and which did not. The new generation of CLSM's (e.g.Lecia SP2) allow collection of data of specific wavlength's chosen through the software and if you collect the image on sequential - basically for example collect UV excited, then 488 and 633 nm data and then merge the data. There is obviously stretch boxes to adjust the image colours collected for each wavlength collected but this process is very subjective and is not mathematical as in flow, the nearest approximation is to try and remember very roughly what the image looked like (very difficult with Cy5 as only part of bright epitopes can be observed by eye) and adjust the colours accordingly. I hope this is of some help as I am more into flow than microscopy, there is a US company called "Universal Pixel" that sells Mac software which seemed very good, although I'm sure there are a lot of other alternatives available. Mac's are a better platform for doing this as a lot of PC software pixelates the images when trying to enlarge the image. Best Regards Gary Warnes, FACSLab, Cancer Research UK London, UK mr wrote:- We are starting a multicolor project by microscopy (please, don't hurl any epithets at me: I'm a true-blue-flow guy... just dabbling a little in the obscure arts). Can someone recommend decent software (preferably Mac) that can do basic image processing that can load multiple images (taken with different filter sets), and then compensate them to give pure fluorochrome images? -- Dr. Gary Warnes FACS Laboratory Cancer Research UK 44 Lincoln's Inn Fields London WC2A 3PX P: 020 7269 3394 F: 020 7269 3100 mail to:gary.warnes@cancer.org.uk Charity No: 1089464 OCTOBER IS BREAST CANCER AWARENESS MONTH To find out more visit www.cancerresearchuk.org
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