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We have been using a prepared diatom slide from Carolina Biological
as a quick demo for our confocal (Leica SP1). According to Carolina,
they stain the mixed diatoms slide "2 different ways and then mix
them together. Some of the diatoms are stained with Harris
hematoxylin and fast green, and others are stained with Harris
hematoxylin and phloxine. "
When the sample is illuminated with blue light (488nm), we get very
little fluorescence. When the same sample is illuminated with green
light (543nm), we get very bright fluorescence in the chloroplasts.
The surprise is that when we illuminate with red light (633nm), we
get dramatic fluorescence of the cell walls in the far red (about 700
and beyond).
Can anyone suggest why we see this long wave fluorescence?
Joel
--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
[hidden email]
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs
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