RE: [Histonet] Blood smear
|From:||"Monson, Frederick " |
Dear Zhiging Xing,
I have never lost even a part of a blood smear, or a mesentery
spread, from a clean slide after proper drying. Although it has long
been clear that slides, out of the box, are considered clean, in fact,
slides in an opened box can pick up all kinds of 'bad' karma in a
functioning laboratory. Thus, throughout my career, I have always kept
a scrupulously cleaned jar filled with 70% ethanol into which I have
placed slides, out-of-the-box, that I have cleaned with Lava soap
(between thumb and (sore) fore finger (left hand!)), then, rinsed in a
stream of running hot water, immersed in cold water in a beaker (then
run for 5 minutes), and finally 5x rinses with deionized water before
being relocated into the 70% ethanol jar, one at a time (with gloves
Needless to say, such practices have conspired to keep me from
earning a living in histotechnology, but I have NEVER lost even a part
of a blood smear to such slides after the smear was FULLY air dried!!
Even when I have rushed, only the thicker parts of the smear are at risk
of 'falling' off. The feathered edges, bless them, always remain intact
for quality analysis, and the basophil(e)s never disintegrate unless
they are weak.
Cheers to you and to all, and please forgive my rip-arte'.
Frederick C. Monson, PhD
Light, Electron, X-Ray and Scanning Probe Imaging and Analysis Center
Large Scientific Instrument Core
Geology, West Chester University
S. Church St. and W. Rosedale Ave.
West Chester, PA, 19320
Knowledge is the key to happiness. Ignorance might be the key to
[mailto:firstname.lastname@example.org] On Behalf Of Xing,
Sent: Tuesday, March 14, 2006 7:57 PM
Subject: [Histonet] Blood smear
I am trying to do X-gal staining on mouse blood smears. But the blood
film keeps coming off the slide. I believe the smear is thin enough.
After making the smear, I dried it overnight at RT or 2-3 hours in
vacuum. Then if I fix it with 100% methanol, the slide would be OK, the
blood doesn't come off during the staining, but I don't get any X-gal
stain. I tried fixation with acetone for 20 min, the blood stayed on the
slide when in acetone, but it came off when transfered to staining
solution. I also tried fixation with glutaraldehyde or PFA, the blood
just comes off even in the fixative solutions. In the begining, I was
using Superfrost plus slides. Somebody on the histonet said the blood
smear might not like the plus slides. So I changed to the regular
slides, but the problem remains.
Can anyone give me some suggestions? Thanks.
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