[Histonet] iron stain on bone marrow-SOLUTIONS

[Gayle Callis]

Have used this rule with CONCENTRATED acid going into water to prevent
spattering plus creating heat.  However, mixing 20% HCl (more water than
acid) with 10% potassium ferrocyanide didn't make any difference.  I am
sure I have mixed these both ways, and I never noticed spattering.
Histotechnic textbooks do not address the safety issue here nor state any
sequence of events.  

The old poem learned in my college chemistry class Chem 101 back in the
dark ages  was applied to concentrated stock acids, and still used  - goes: 

"Always do what you oughter, pour the acid in the water!" 


At 01:35 PM 3/18/2004 -0500, you wrote:
>"Remember the order: acid to water"
>
>-----Original Message-----
>From: Bonner, Janet [mailto:Janet.Bonner <@t> FLHOSP.ORG] 
>Sent: Thursday, March 18, 2004 12:56 PM
>To: 'Wendy England'; histonet <@t> pathology.swmed.edu
>Subject: RE: [Histonet] iron stain on bone marrow-SOLUTIONS
>
>
>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
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)