They can be seen in dry slices, right after slicing. I wonder if they are bone pieces but have no idea of why they specifically bind to the GL. Any idea of how to get rid of it?
I have exactly the same problem in mice, some atofluorescent deposits in the GL especially between the 2 ob. I never found the origin of it.
Best,
In my experience these are present without any staining or even fixation procedure.
Nicolas is right. It is there before any staining. I thought for a while they were cristals from the secondaries or Dapi. But no, they are there right after slicing.
There is an autofluorescence in the golmerular layer that can be excited with 740 nm multiphoton microscopy in fresh slices. It overlaps well with the dopamine neurons. The emissions spectrum is from about 425 to 550 nm with a peak of about 460 nm. This overlaps with NADH. I am not sure if it is visible with single photon excitation. What are excitation/emission filters are you using?
Makes all the sense in the world that could be cathecol-related autofluorescence. The Hillary and Falk method required aldehydes, but a specific environment could allow some carbon rings to fluoresce intensely. Is the animal old? These old geezers can have some protein deposits such as lipofuchsin, or 'glycated' proteins or lipids droplets from lipid accumulating cells. Send some photos for me Eduardo, I got curious.
Here it is João. It is a low magnification image on an epifluorescence microscope. These guys were around 3 months old. I can send better pictures when I get back to the lab next month.
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