Week 3

Dr. Dimos

Ex.11 Simple Staining

Use a dye to increase contrast

Basic dyes attracted by negatively charged bacteria:
Methylene blue
Crystal Violet

Swab cheeks cells and stain w methylene blue

Cheek cells are large irregular shaped

epithelial cells with a very obvious nucleus

Ex.12 Negative Staining

Used to study morphology of bacterial cells

Can see external structures such as capsules
Cells appear transparent against a dark background
Acidic dyes repelled by bacteria:
India ink
Nigrosin, nigrosine
Swab gumline for bacteria and stain with nigrosin

Ex.13 Capsular Staining

Capsule or slime layer surrounds the bacterial cell

Used to attach to surfaces
Used to evade immune system
Capsular staining combines simple staining and negative
staining
Capsules look like halos surrounding colored cells against a dark
background
Look at a prepared slide of a capsular slide

Ex.15 Spore Staining

Endospores of bacteria vegetative

states
Bacillus and Clostridia form
spores
Extremely resistant to heat and
chemicals
Therefore, also resistant to most
stains
The stain malachite green can
penetrate endospore if applied with
heat
Look at prepared slide of a spore
stain

Ex. 8 Aseptic Technique

Purpose
Prevent cross contaminating cultures with other undesired
organisms
Prevent contaminating self or others
Prevent leaving behind contaminated areas

Ex. 9 Pure Culture Techniques

Naturally microorganisms exist

as mixed cultures
Pure culture allows the study of
an individual organism
Pure culture methods
Streak Plate
Pour plate
Both methods allow the
isolation of a single colony
all identical to the original
cell

Ex.10 Smear
Preparation
Goals
Cells must adhere to
slide
Avoid shrinkage of
cells
Thin smears - no
clumps!
Prepare four smears from
liquid culture:
1. Gram negative culture
(E.coli)
2. Gram positive culture
(S.epidermidis)
3. Mix of gram positive and
gram negative

Ex.14 Gram
Staining

Ex.14 Gram Staining