| From: | "Glenn Smith" |
In a message dated 10/24/2003 3:01:17 PM US Mountain Standard Time, Bonnie.P.Whitaker@uth.tmc.edu writes:
> I have a question for you guys: A researcher here in OB/GYN is doing rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse fallopian tube... he thought he was having a staining problem, but has determined that his tissue is auto-fluorescing. The tissue is frozen in OCT and fixed in acetone/methanol. What can he do to quench this? <
A couple more thoughts Bonnie...
Rhodamine is excited with 488nm (blue light). I am assuming the system used in this case employs such a light source. Most organic material fluoresce under 488nm excitation. The best option is to use a narrow bandpass filter (20-40nm bandwidth centered about the peak emission wavelength of the dye). This will not completely eliminate autofluorescence but it will greatly reduce it. Another possibility is to image the tissue before and after staining to see the difference. Then could subtract one image from the other...