Hi Manisha,
The technique is very simple and give very consistent results. I have provided a reference below from my previous institution:
Prior bleeding enhances the sensitivity of peripheral blood and bone marrow micronucleus tests in rats. Mutagenesis (2007) 22 (4): 287-291.
This should work with mice also.
Regards,
Kamran
Manisha,
original protocol is here Schmid W. The micronucleus test. Mutat Res1975 Feb;31(1):9-15
fluorescence stain described here Hayashi M, Sofuni T, Ishidate M, Jr. An application of Acridine Orange fluorescent staining to the micronucleus test. Mutat Res1983 Jun;120(4):241-7
and here Tinwell H, Ashby J. Comparison of acridine orange and Giemsa stains in several mouse bone marrow micronucleus assays--including a triple dose study. Mutagenesis1989 Nov;4(6):476-81.
hope it helps
Harvest bone marrow cells from demure in chilled sodium citrate solution(1%) ,centrifuge the cell suspension at 1000 rpm for10 min, resuspendcells in few drops of sodium citrate solution, and prepare a smear .Air dry the smear fix in methanol for 10 min ,stain in 4% Giemsa prepared in phosphate buffer for 30 min ,rinse in water and observe the presence of micro nucleus in enucleated erythrocytes.
Hi Manisha
Perhaps you should review the papers from Ruth Wilkins's lab at Health Canada. They have been doing beautiful (IMHO) work with the ImageStream (Amnis/MerckMillipore) and the far-red cell permeant DNA probe DRAQ5 (BioStatus) in blood samples for micronucleus detection (aka genotoxicity in drug development) in the event of a mass radiological hazard event like the Fukushima Daiichi nuclear disaster (Mar 2011). The Google Scholar link here will get you to all the papers they've published on this. This should be amenable to bone marrow with the caveat of how you manage the sample prep to have a single cell suspension, as far a possible.
I hope that's useful info.
Roy
http://tinyurl.com/j9djzuv
http://tinyurl.com/h4lo4hk
The pioneer protocol published by Schmid W. The micronucleus test. Mutat Res1975 . It has also incorporated in OECD genotoxic guideline for chemical testing. It is very simple and cost effective technique to evaluated the genotoxicity as per your applications.
I have experience and expertise in this directions and published original articles in the term of radiation induced genotoxicity as well as its amelioration in bone marrow of gamma exposured to mice by administration of radioprotectors.
Hey Manisha, you have to go following link and get the easy way to proceed further for MN Assay in Bone marrow
Article Sesamol ameliorates radiation induced DNA damage in hematopo...
Article Sesamol attenuates genotoxicity in bone marrow cells of whol...
Article Evaluation of sesamol-induced histopathological, biochemical...
Please fell free to further technical support......
Regard
Arun
Respected scientis, I staibed mouse bobe marra with May Gruwalds /Giesama stain whjch prepared but not getting clearity of detecting micronucleus in PCE NCE so kinfly help me on how to pre working solutuin stsining protocoks.