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acetocarmine staining of microspores

Constantine A Lapasha calfor at unity.ncsu.edu
Thu Apr 7 07:14:12 EST 1994

okellyc at megasun.BCH.UMontreal.CA (Charles J. O'Kelly) writes:

>In article <bucc0003.765611484 at gold>, bucc0003 at gold.tc.umn.edu (Paul A Bucciaglia) writes: 
>|> I've been staining tobacco microspores (about 3-5 days post tetrad) with
>|> acetocarmine to follow microspore mitosis.  Using the std. 1% carmine
>|> in 45% acetic acid on both fixed (ethanol/acetic acid) and unfixed antehrs
>|> gives me bright red cytoplasms that obscure any sign of a nucleus.  i dug
>|> up a reference from 1937 (Maheshwari, Stain Tech. 12#2) which suggested
>|> a few drops of chloral hydrate to clear the cytoplasm.  "ahh, an archaic
>|> name for HCl" i thought so i added a drop of 1N HCl.  This did help some
>|> as the nucleus was just barely visible and the cytoplasm cleared somewhat.
>|> So i checked the Sigma catolog and was surprised to find that chloral 
>|> hydrate is a nasty chlorinated hydrocarbon that is (was?) used to make DDT.

>Chloral hydrate was also the active ingredient in the Mickey Finn (knockout drops
>added to alcohol in all the old film noir mysteries).

>Given the above, why not relax and learn to use chloral hydrate safely?

>|> paul bucciaglia

>Charley O'Kelly

the other problem we ran into when checking into using chloral hydrate
as an ingredient in a clearing solution was that chloral hydrate is a
controlled substance -- the permits and paperwork involved are a giant
hassel - to say nothing of the worries of storing a current "street drug"
in the lab --- we decided to work around the problem rather than go through
the federal and state controlled substance permit routine

calfor at unity.ncsu.edu

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