[Histonet] RE: Hoechst staining method

From:Melanie Black

Hi Tracey

Some years ago, I looked into fluorescent nuclear counterstains, 
including dapi, propidium iodide and HOECHST. Heochst is also known 
as bisBenzimide.

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)

Melanie Black

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.


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