How to use the DAPI for detect autofluorescence lipofuscin in C.elegans? Or any methods?
I know, I know…you’re thinking (not the original poster of course) why don’t you just leave it…but I’m a caring-sharing kind of guy and ill-conceived, muddled questions like this one make me want to help even if, as in this case, the cause is lost…sigh. (Perhaps I should report myself to myself for being so dumb?)
So…first…just for the incredulous amongst us, you mean the DAPI emission filter right, you haven’t already gone ahead and blown your lab’s budget and bought DAPI itself?
Second, for those still with a will to live…you mean autofluorescence originating from the gut granules? Then the lipofuscin tree is the wrong one to bark up, see;
Babu, P.(1974).Biochemical genetics of Caenorhabditis elegans. Mol.Gen.Genet. 135, 39–44.doi:10.1007/BF00433899
Bhat, S.G.,and Babu, P.(1980). Mutagen sensitivity of kynureninase mutants of the nematode Caenorhabditis elegans. Mol.Gen.Genet. 180, 635–638. doi:10.1007/BF00268072
Siddiqui, S.S.,and Babu, P.(1980). Kynurenine hydroxylase mutants of the nematode Caenorhabditis elegans. Mol.Gen.Genet. 179, 21–24. doi:10.1007/BF00268441
the cat is probably in the anthranilic acid glucosyl ester ‘tree’ (see Coburn et al., (2013) PLoS Biol. 11:e1001613. doi:10.1371/journal. pbio.1001613.
quote: ‘Or any other methods?’ ???
thank you for your help,but I still have not some understand.
i see some papers detect the Lipofuscin fluoresces directly,not need stain.
other,e.g.,Measuring Caenorhabditis elegans life span on solid media.J Vis Exp. 2009 May 12;(27). pii: 1152. doi: 10.3791/1152.Do i need dapi？
no, you don’t need dapi, but you need a microsope equipped with epifluorescence optics and a standard DAPI filter cube.
that steve h is a great guy!