[Histonet] iron stain on bone marrow-SOLUTIONS

[NICK KIRK]

Hi there
 
In my experience it doesn't matter which way round you add the solutions, plus I think the HCl solution you are using is far too strong.
The Perl's prussian blue technique I use uses 2% HCl and 2% Pot ferro and works excellently every time even on marrows.
We incubate for about 15 minutes.
 
Hope that helps
 
Nick Kirk
Histopathology
Hinchingbrooke Hospital
Huntingdon
England
 
 

"Bonner, Janet" <Janet.Bonner <@t> FLHOSP.ORG> wrote:
The 20% Hydrochloric acid solution should be added to the 10% Potassium
ferrocyanide solution - (not 10% potassium ferrocyanide to 20% Hydrochloric
acid solution) when making the 1:1 mixture.


-----Original Message-----
From: Wendy England [mailto:weneng <@t> hotmail.com]
Sent: Wednesday, March 17, 2004 3:28 PM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] iron stain on bone marrow


Thanks again to everybody who gave me the information about this stain!
Here comes another question: I couldn't see much iron on my slide, only 
about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 
min, then in freshly mixed stain solution(equal part of 20% HCl and 10% 
potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin 
after wash. Then wash, air dry then mount.

I was told there should be much more iron. I feel bad now. Could any body 
give me advice or could I ask if it's possible due to the bone marrow 
preparation since we indeed saw some spots. I didn't prepare the sample.

Thanks and look forward to hear from you there!

Wendy

_________________________________________________________________
FREE pop-up blocking with the new MSN Toolbar - get it now! 
http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/


_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


The information contained in this message may be privileged and/or
confidential and protected from disclosure. If the reader of this message
is not the intended recipient or agent responsible for delivering this
message to the intended recipient, you are hereby notified that any
dissemination, distribution or copying of this communication is strictly
prohibited. If you have received this communication in error, please notify
the sender immediately by replying to this message and deleting the material
from any computer. 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet