Re: [Histonet] Genetics anyone?

From:Hadi Yaziji

More on ploidy:

Using "stain" is most likely referring to Feulgen stain. With image 
analysis it becomes more accurate, but still semiquantitative. Success 
depends on the image analysis software and experience of operator.

Flow cytometry requires grinding either fresh tissue or 50 microns of 
the tissue block. It's certainly more quantitative.

FISH is the enumeration of chromosome copies. On tissue sections (using 
2-6 micron sections), chromosome centromere probes of more than one 
chromosome (preferably 3) are used in the same kit. This is as close to 
quantitation as it gets. It is accurate, and relatively easy to use.

Cytogenetic spread is the real quantitative test, but it requires fresh 
tissue sample and growing it in cell culture, just like amniocentesis 
testing for fetal chromosomal abnormality. Software image analysis 
assistance is also available for the last two.

As colleagues already indicated, your researcher is most likely 
implying "Fuelgen" stain.


Hadi Yaziji, M.D.
PhenoPath Laboratories

On Nov 6, 2003, at 8:20 AM, Morken, Tim - Labvision wrote:

> Cheryl, As noted by others, ploidy can be determined using the feulgen
> stain. While a simple bright-field examination can show differences in 
> stain
> intensity related to ploidy that would be a very rough guess as to 
> actual
> ploidy differences. More exact analysis requires either an image 
> analysis
> system (several on the market) or flow cytometry.
> Here is a concise explanation of one method of ploidy analysis:
> Tim Morken
> Lab Vision / NeoMarkers
> -----Original Message-----
> From: Cheryl Crowder []
> Sent: Thursday, November 06, 2003 5:47 AM
> To: Histonet
> Subject: [Histonet] Genetics anyone?
> Good morning - This questions was put to me yesterday and, knowing
> little about genetics, I'm stumped.  Can any of you help me.  This
> researcher is collecting samples which he says are either diploid,
> triploid or tetraploid.  He states that someone told him there is a
> stain techniques that "would distinguish the 'ploidy' by the intensity
> of the stained tissue".
> Have any of you heard of such a thing or know someone I can contact for
> this "unusual" question?  Thank you, in advance.  Cheryl
> Cheryl Crowder, BA, HTL(ASCP)
> Chief Technologist
> Anatomic Pathology
> Department of Pathobiological Sciences
> School of Veterinary Medicine
> Louisiana State University
> Skip Bertman Drive
> Baton Rouge, LA  70803
> 225-578-9734
> FAX:  225-578-9720
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