To teach new users the basics of how our confocal works (before we get to
working with their samples) I use a kimwipe that has been marked with
several different fluorescent highlighters. I mount a piece of the kimwipe
to a slide with permount under a #1.5 coverslip and it lasts almost forever.
It's incredibly bright, moderately thick (~90um), cheap to make, and 3D
rotations using a 10x objective show the fibers very nicely. This teaching
slide idea was inspired by my colleagues "up the road" at AZ State
University with their Paper project < http://paperproject.org/>.
The kimwipe slide is not very effective to show two different wavelengths,
but it lets me talk about spectral bleedthrough/artifactual colocalization
issues and why the longer wavelength channel is usually noisier (PMT gain is
often twice that of the shorter wavelength).
The Invitrogen Fluocells slides (no commercial interest) are beautiful for
demos, but they are expensive.
H&E stained histologic sections fluoresce and I've been told that diatoms
(good for demonstrating DIC if your confocal has all the parts for it) are
autofluorescent.
Have fun.
Doug
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA
office: AHSC 4212 email: [log in to unmask]
voice: 520-626-2824 fax: 520-626-2097
http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Jonathan M Krupp
Sent: Friday, November 14, 2008 9:46 AM
To: [log in to unmask]
Subject: Practice samples
Hi:
This is a pretty basic and simple question. I need some
advice about rounding up some samples to use in an
elementary light microscopy class that includes basic
fluorescence.
I am at a community college and my background is in EM and
brightfield LM. I have a passing familiarity with confocal
et al but not enough to know much. I maintained a
confocal, but never ran experiments or prepared much in
the way of samples. Now I have to get something together
to demonstrate the fundamentals of fluorescence to
students in a new job.
I have been relying on chlorophyll autofluorescence up til
now, but would like to add anything that would be easy to
do. We have a simple scope with filters for FITC,
rhodamine, and DAPI, I think. What would be some fool
proof, easy to get samples to try?
In addition, I would eventually like to add some kind of
confocal experience to this class, any ideas about where
to find an inexpensive system would be great.
Thanks
Jon
San Joaquin Delta College
Stockton, CA 95207
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