[Histonet] RE: Hoechst staining method
Hi Tracey
Some years ago, I looked into fluorescent nuclear counterstains,
including dapi, propidium iodide and HOECHST. Heochst is also known
as bisBenzimide.
Method:
Mix 1mg bisBemzimide (Sigma B2883) with 1ml Dist water. Take 4.0
micro litres of this stock and dilute with 1ml PBS pH 7.4.Apply to
sections for 10 mins, wash 3 x in PBS. Fluorescence is bright blue,
viewed with a UV filter 340 nm.
Stain reference:
Latt,SA and Stetten GJ. Histochem. Cytochem 24,24 (1976)
Araki, T et al, Histochem. 87,331 (1987)
Regards
Melanie Black
Hi,
We are doing an immunofluorescence research project for podocytes.
The urine cytospins are to be counterstained with Hoescht. We have
never used it - can someone recommend a concentration, diluent and
staining time that we could start with. Thanks in advance.
Tracey
--
Cardiovascular Research Unit
Div. of Cardiothoracic Surgery
Chris Barnard Building
University of Cape Town
Anzio Road
Observatory
7925
Republic of South Africa
Tel +27 21 406-6589
Cel +27 82 469-3352
Fax +27 21 448-5935
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