We try to fix bacteria (E. coli) on glass slides for staining the with different antibodies and membrane stains. Most protocols I found were for living cells, which is not what we need.
Does anyone has some tips?
Maybe you can use simple Poly-L-Lysine or gelatin-chrom-alum coated slides?
Fixation conditions of 2-4% paraformaldehyde in 0.1M phosphate buffered saline for 12 - 24h at 4°C are common for IHC.
Are the bacs in solution or on agar?
If they are in in a liquid, you can produce a "cell-block". Spin the liquid down to concentrate the bacs on the bottom and take off the supernatant. Take the bacs in a few drops of formalin.
Warm a small amount of agar until liquid and squirl the bac-solution into the liquid agar.
This can be done in an Epi. Spin the agar to concentrate the bacs in the lower area.
Let cool the agar to make it solid. Take it out of the Epi (eg. with the help of a thoothstick) and fix it over night in formalin.
Then process the agar-block in a Histo-cassette to make a regular paraffin-block.
From this block you can section many slides to stain your IHC.
If the bacs are on agar, just take some colonies and squirl them into the warm liquid agar. and so on.
This paraffin--slides can be treated like any other FFPET-slides.
For bacs in liquid it would also a opportunitiy to use a "Cyto-Spin" centrifuge. You can find such an instrument in a Cytology-lab. If you use adhesive slides with the Cytospin, you can also do your IHC with no lost of bacs. It would be of advantage to fix the glass-slides in formalin.
If my explanation is not clear enough, look for Cell-block preparation in the internet.
hope this helps.
- Badges
- Science method