Agar slide preparation

Protocol name: Agar slide preparation
Prepared by: Monica Ponder
Date Created/Revised: 5/27/03
Purpose: To allow visualization of
microbial cells by light microscopy
so that cells are adhered to the
surface and produce less shadow
Reference(s): Adapted from advice of Frank
Prepare cultures to a very low density, 0.1- 0.2 OD 600, 50 ul spotted onto slide
To prepare agar coated slides:
Prepare 0.75% Nobel agar (0.375g per 50 mls of MilliQ). Sterilize 25 minutes.
To microscope slide add 0.250 mls of melted agar to end of slide and drag another slide
across at a 45° angle so that a very thin film of agar is left on the slide. Allow slides to
dry under UV light to prevent contamination. Store in sterile Petri dish until ready to use.
A good photograph should have at minimum 6 bacteria but no more than 30 per field and
have all firmly affixed to slide. Streaks are not good.
You should also include a size marker for published photos.