Unit 1 · Pathogenic bacteria

The Gram stain, step by step

One 5-minute stain splits most bacteria into two camps. The secret is the cell wall: thick peptidoglycan holds purple dye, thin peptidoglycan plus an outer membrane lets it wash out.

1000× · oil immersion
Gram-positive cocciGram-negative rods

Zoomed into the cell envelope. Cytoplasm is at the bottom, the outside world at the top.

Won't Gram stain well
Mycobacterium (TB)
Waxy mycolic acid wall repels the dye. Use the acid-fast (Ziehl–Neelsen) stain: they show up red.
Mycoplasma
No cell wall at all, so nothing to stain. Also why penicillin can't touch it.
Treponema (syphilis)
Too thin to see. Use darkfield microscopy.
Chlamydia, Rickettsia
Live inside our cells.