Re: FISH measurement

Posted by Mark Cannell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/FISH-measurement-tp3480943p3483125.html

Due to the non symmetrical PSF I don't think this is directly possible.
There is a solution tho' please see:

Soeller, C. & Cannell, M.B. (1999) Examination of the transverse tubular
system in living cardiac rat myocytes by 2-photon microscopy and digital
image-processing techniques. Circ. Res. 84: 266-275

Note that  this blurring method does not depend on imaging modality but
makes the effective PSF spherical which is what you need.

Cheers Mark


Carl Boswell wrote:

> Hi all,
> I have a user who wants to measure the volume, not intensity, of the
> FISH-labeled region in a z-series.
>
> Any issues with the following proposal, besides using a histogram to
> get pixel numbers?
>
> 1. Threshold the FISH signals for control and experimental Z-Series
> images with identical conditions.
> 2. Create a histogram for each thresholded image and use total pixel
> number as
> the FISH signal volume.
>
> Thanks,
> C
>
> Carl A. Boswell, Ph.D.
> Molecular and Cellular Biology
> University of Arizona
> 520-954-7053
> FAX [phone]