I am currently doing micronucleus assay in cancer cells.
I am wondering on how to calculate the frequency % cells with micronucleus from the images I collected and the best approach to calculate the error (standard deviation) and significance.
Some image-based analysis use the ratio of the positive/negative cells per image, some other using the total cell number instead of ratio per image. Which method should I use for micronucleus assay?
A standard method is to count manually 1000 cells under oil immersion and determine % with micronuclei.
see attached paper
Agreed with James Leigh sir... a standard methodology is to count the minimum 1000 cells per slide to calculate the micro nuclei frequency. Most of MN guidelines have suggested the same.
the attached documents will definitely help you.
Thank you for your responses, Sir James Leigh and Arun Kumar
I have read several articles about these methods related to genotoxicity, including one published by Fenech (2007). My further question is, will these guidelines applicable for different purpose of experiment (we are not measuring toxicity level, but chromosome segregation errors), and fluorescence microscopy?
The nature of these guidelines is quite different from ours since we do not use the cytokinesis block, nor counting the bi/multi-nucleated cells.
If we use the 1000-cells counting method, how do we ensure the objectivity of the calculation without using the double-blind method?
Really appreciate your answers.
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