[Histonet] RE: Hoechst staining method

[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.

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

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