while working on the concept of distinguishing the live and dead cells by dye exclusion principle, widely TB, PI and Methylene blue (MB) are in use.
In context to commercially available ready to use MB stain, what % of dye is preferred for the analysis ? Does it requires a dilution or can be used directly?
Dear Bhavya G, as usual, it might depend on the "cells" you try to stain - Usually concentrations of 0.1 - 1% (w/v) MB-dye will be used (so it would be helpful to know the kind of cells you are going to analyse).
cf: e.g. (not exhaustive!):
A quick method for estimating the percentage of viable cells in a yeast population, using methylene blue staininghttp://www.westerdijkinstitute.nl/BioloMICSNews.aspx?Rec=5639
For comparison of several methods of intra-vital stains e.g.:
Article Comparison of methods used for assessing the viability and v...
=http_s://onlinelibrary.wiley.com/doi/full/10.1111/1567-1364.12202 (Open Access at Journals website, when copying and pasting the latter URL into your browser: DELETE the "underline"-character _ in between "http" and "s://....", for the pdf (vide: also delete the added underline-character) see:
http_s://onlinelibrary.wiley.com/doi/epdf/10.1111/1567-1364.12202
also:
http://www.protocol-online.org/biology-forums/posts/30995.html
or:
http_s://www.ncbi.nlm.nih.gov/pubmed/10856773 (delete underline-character) =
Article Simple detection method for distinguishing dead and living y...
".... Methylene blue, phloxin B, rose bengal and trypan blue at concentrations of 5-10 micromol l(-1) were shown to stain non-viable cells ...."
Use of Immunofluorescence and Viability Stains in Quality Control by M. J. CHILVER et al, 1977:
http://www.agraria.com.br/extranet/arquivos/agromalte_arquivo/imuniflorescencia_para_determinacao_da_vitalidade_ing_-_ing.pdf
or:
A MICROSCOPIC METHOD OF DISTINGUISHING DEAD FROM LIVING BACTERIAL CELLS by KNAYSI G., 1935 see:
http://jb.asm.org/content/30/2/193.full.pdf = in ResearchGate (when trying to paste the URL:
http_://jb.asm.org/content/30/2/193.citation (NB: before pasting URL into your browser, delete the underline character in between http and ://...) otherwise -as follows - you end up with:
Article A Microscopic Method of Distinguishing Dead from Living Bact...
Best wishes and regards, W.M.
Dear Bhavya G, as usual, it might depend on the "cells" you try to stain - Usually concentrations of 0.1 - 1% (w/v) MB-dye will be used (so it would be helpful to know the kind of cells you are going to analyse).
cf: e.g. (not exhaustive!):
A quick method for estimating the percentage of viable cells in a yeast population, using methylene blue staininghttp://www.westerdijkinstitute.nl/BioloMICSNews.aspx?Rec=5639
For comparison of several methods of intra-vital stains e.g.:
Article Comparison of methods used for assessing the viability and v...
=http_s://onlinelibrary.wiley.com/doi/full/10.1111/1567-1364.12202 (Open Access at Journals website, when copying and pasting the latter URL into your browser: DELETE the "underline"-character _ in between "http" and "s://....", for the pdf (vide: also delete the added underline-character) see:
http_s://onlinelibrary.wiley.com/doi/epdf/10.1111/1567-1364.12202
also:
http://www.protocol-online.org/biology-forums/posts/30995.html
or:
http_s://www.ncbi.nlm.nih.gov/pubmed/10856773 (delete underline-character) =
Article Simple detection method for distinguishing dead and living y...
".... Methylene blue, phloxin B, rose bengal and trypan blue at concentrations of 5-10 micromol l(-1) were shown to stain non-viable cells ...."
Use of Immunofluorescence and Viability Stains in Quality Control by M. J. CHILVER et al, 1977:
http://www.agraria.com.br/extranet/arquivos/agromalte_arquivo/imuniflorescencia_para_determinacao_da_vitalidade_ing_-_ing.pdf
or:
A MICROSCOPIC METHOD OF DISTINGUISHING DEAD FROM LIVING BACTERIAL CELLS by KNAYSI G., 1935 see:
http://jb.asm.org/content/30/2/193.full.pdf = in ResearchGate (when trying to paste the URL:
http_://jb.asm.org/content/30/2/193.citation (NB: before pasting URL into your browser, delete the underline character in between http and ://...) otherwise -as follows - you end up with:
Article A Microscopic Method of Distinguishing Dead from Living Bact...
Best wishes and regards, W.M.
You're welcome and your recommendation(s) are appreciated....
Naturally the previously made statement is not the last wisdom with regard to your question. There are several methods (using Mehylene blue) which can be applied.... to ???cells???, "cell suspensions???" ...either hydrous or (obviousely far better, in a phosphate buffered solution at pH 4.6....
0.01 -0.02% MB, 1:10:000 aqueous, or 1:10,000 aqueous solution pH 6.8
Have you considered "SEARCHING" the ResearchGate Archives [ either in 'Publications' or also -naturally ! - 'Questions' ] for keyword(s) or even phrases (FULL TEXT SEARCH) e.g. :
| Methylene Blue vital supravital intravital stain (staining) dead cells live cells | ....you might be surprised what is already saved in ResearchGate !. If you don't know how to do, just ask for it...and it will be explained to you....Best regards: W.M.
Honestly added from my collection (recipes, "sources" "EM-work", etc.): YEAST: "dead or alive....!"
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