Hello,

You can do negative staining with transmission electron microscopy.

You need TEM grids coated with formvar for example, negative stain (1% uranyl acetate for example), fine tweezers, gridbox.

-Make your bacteria (suspension) adhering on the grid.

-Remove excess of liquid with filter paper.

-Contrast with for 30-60 seconds.

-Remove excess of liquid with filter paper.

-Let dry.

-store in gridbox.

for more details look at: https://www.researchgate.net/post/Hello_I_am_trying_to_visualize_my_liposomes_under_TEM_Can_anyone_help_me_with_details_of_PTA_staining2

It is about liposomes, but it is the same way for bacteria.

If you want to do quantification, your bacteria suspension should not be to concentrated because you need isolated bacteria.

You can use ImageJ in semi automatic mode to quantify the number and length of pilies.

Hello Nicolas Barois ,

Thanks for answering

When thinking quantification, always take pictures at the same magnification.

If you are interested, I have a semi-automated macro for ImageJ designed for bacillus cereus, made some years ago, to quantify bacteria and exosporium surface (using threshold), then number and length of pili (using polyline). it works with one bacterium per image.

The macro and its notice are in french, but you are in Quebec, Canada, shouldn't be a problem.

Nicolas Barois i really appreciate your help

it would be nice to read that macro