I used to stain flagella using the Leifson's method, which can be found in
any basic microbiology methods manual.
The secret, in my experience, was not only using new glass slides, but also
treating them with chromic mixture (I believ it is a mixture of sulfuric
acid+potassium dichromate) for 5 days. Then, rinsing them thoroughly with
distilled water. DO NOT WIPE THE SLIDES. Then, dipping them in ethanol and
As far as the cultures, we grew them in TSA slants for 18 h, at 35°C, and
tempered sterile saline solution at 35°C before adding 2-3 ml onto the
slant, rubbing gently with the pipette to make a suspension DO NOT VORTEX.
The smears were prepared taking a little of the bacterial suspension with a
sterile pasteur pipette, then adding one drop onto an end of the glass
slide, then tilting the slide to allow the suspension to run over the lenght
of the slide. Let air-dry (DO NOT HEAT FIX) and cover with freshly made
Leifson staining solutions. Use all reagents of the highest purity possible.
even NaCl was an issue.
It's been along time since we worked with flagella staining. New or better
methods may be around and I don't know.
I hope this helps.
"Debra L. Swanson" wrote in message <363a3b450036002 at mhub2.tc.umn.edu>...
>Does anyone have experience with a successful protocol for staining
>>Debra Swanson, Assistant Scientist
>University of Minnesota
>Minneapolis, MN 55455
>swans061 at maroon.tc.umn.edu>