Re: Re; Nuclear volume, ploidy and Stain stochiometry

Posted by Derek Ingram on Feb 18, 2009; 9:38pm
URL: http://imagej.273.s1.nabble.com/Re-Nuclear-volume-ploidy-and-Stain-stochiometry-tp3693629p3693632.html

First of all I would like to thank everyone for their input and interest
regarding my initial posting.  I should elaborate on my initial
question. Annually the company I work for evaluates ploidy via Propidium
Iodide and flow cytometry. Alternatively we would like to develop a
technique to determine ploidy in various strains of salmonids using
IMAGEJ and preferably unstained blood smears. I would be more then happy
to forward some journal articles to whomever is interested regarding
ploidy determination through means of nuclear measurement. It has been
done before. The reason I am trying to utilize IMAGEJ software is to
speed up the processing of samples. I have no problem measuring total
cell measurements with other software. The issue I have with IMAGEJ is
isolating the nucleus, I'm sure this should be a simple task, but due to
my inexperience with the software I am having some issues. Again I would
like to thank everyone who is responding.

Derek Ingram

-----Original Message-----
From: ImageJ Interest Group [mailto:[hidden email]] On Behalf Of
Alton D Floyd
Sent: Wednesday, February 18, 2009 10:49 AM
To: [hidden email]
Subject: Re: Re; Nuclear volume, ploidy and Stain stochiometry

For autoradiographic grain counting, the type of emulsion, developer
used, and method of development all influences the grain size and
morphology.  When this technique was widely used, in the early 1960's,
most investigators used developers that produced tiny, round grains.  As
in any other type of quantitative measurement, the specimen preparation
and grain development must be highly controlled.  

When I was actively involved in this type of work, we found that we had
to create new calibrations each time we used a new batch of emulsion.
The calibration was created by making a series of slides, coating with
emulsion, and exposing for increasing lengths of time.  Then we took
reflectance measurements, and compared these to actual grain counts in
the measured areas.  With such a calibration, we found excellent
correlation between actual counts and reflectance measurements.
Obviously, the measurements were much faster, and permitted studies such
as localization of radioactive tagged materials in serial sections of
organs, such as rodent brains.

I don't remember all of the citations to these type of studies.  I will
check my old citation card file and see if I can come up with specific
citations that will give you a starting point for a citation search.

Al Floyd
23126 South Shore Drive
Edwardsburg, MI 49112
Phone: [phone]
Mobile: [phone]