Quantification of Immunofluorescence Microscopy Results - (May/30/2007 )
Hi friends,
How do you quantify the results of immunofluorescence microscopy? Which software do you use? How do you normalize the results?
I would not quantify immunofluorescence. Its only a method which would give a yes or no type of answer.
I would prefer some other way for quantification.
scolix is right as you cannot guarantee the emission being proportional to the epitope however that aside have a look at 'volocity' we use it here and its good for getting info from pictures (deconvolving also cleans them up) but remember it will allow you to do a lot of stuff which isnt really scientifically valid.
dom
How do you quantify the results of immunofluorescence microscopy? Which software do you use? How do you normalize the results?
you can define ROI´s and compare them in one image if signals are in the linear range of fluorescence; companies provide software to their microscopes to gear the microscope and to collect and analyze the data; microscope-independent is f.i. Bitplane´s Imaris
How do you quantify the results of immunofluorescence microscopy? Which software do you use? How do you normalize the results?
you can define ROI´s and compare them in one image if signals are in the linear range of fluorescence; companies provide software to their microscopes to gear the microscope and to collect and analyze the data; microscope-independent is f.i. Bitplane´s Imaris
How can we compare the results from different days? Is there any way to normalize them?
"How can we compare the results from different days? Is there any way to normalize them? "
many microscopes will allow you to save the parameters (microscope and computer) used on one day and then use them as a standard on the next
dom