Autofluorescence in zebrafish

Yiing Lin ylin at ACPUB.DUKE.EDU
Sat Nov 23 21:20:46 EST 1996


Hello,

I was wondering if anyone has taken a close look at the auto-fluorescence 
that comes from zfish fry?  I was trying to get a more rigorous and 
quantitative sense of what auto-fluor occurs in zfish fry.

(The equiptment that I'm using is a Hg epi-fluor illuminator, using a 
FITC (exitiation filter: 455-500nm; emission filter: 510-560nm) filter 
set.)

With 24hr fry, I see a fair amount of auto-fluor, but that isn't as 
critical, since the bulk of the fry isn't obscured by it.  However, going 
down to 40x, I saw a fair number of small, rather bright spots of green.  
Does anyone have any idea of what these are or could be?

At 48hrs, the autofluor gets rather bright.  Rather interesting patterns 
arise from the pigmentation that's developing.  It's rather difficult to 
optically section the embryo (probably because of the pigmenting?) and 
with the bright autofluor, I wasn't able to find any of the spots of 
green that I saw with the 24hr fry.

I notice that confocal microscopy is rather good at taking out the 
autofluor (probably because of the very tight excitation wavelength and 
the diffuseness of autofluor at a single plane), but I wonder if those 
distinct spots that I saw with the 24hr fry would appear on confocal 
sections?

Has anyone observed the same things, and/or have suggestions about 
dealing with it?

						-Yiing Lin




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