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BACTERIAL STAINING

DEPARTEMEN MIKROBIOLOGI
PROGRAM STUDI PENDIDIKAN DOKTER
FAKULTAS KEDOKTERAN UNIVERSITAS TANJUNGPURA
STAINING
The process of adding a dye to a
bacterial culture
BACTERIAL STAINING
Bacteria cells are almost colorless and
transparent keluar

A staining technique is often applied to the cells to


color them Their shape and size can be easily
determined under the microscope.

Stains combine chemically with the bacterial


protoplasm.
DYES 3,4,5,6,9,10,11,14,yg
ada peptidogikan, no 17, 22
Basic dyepossess a positive charge
Acidic dyepossess a negative charge

Remember, bacteria posses a slight negative charge


Coccus Basil Spiral
5
Diplococcus
Streptococcus
Tetrad
Sarcina
Staphylococcus 6
Diplobacilli

Streptobacilli

Cocobacil

7
Vibrio

Spirillum

Spirochaeta

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Smear Preparation:

Preparation and Fixation of Bacteria for Staining.


Note: dont forget to clean the objective glass

Objective:

To kill the microorganism & fix them to the slide to prevent


them from being washed out during the process of
staining.
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TYPES OF STAINING TECHNIQUES

Simple staining Differential staining


(use of a single stain) (use of two contrasting stains
separated by a decolorizing agent)

For visualization of
morphological Identification Visualization of
shape & arrangement structure

Gram Acid fast Spore Capsule


stain stain stain stain
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Gram Stain:
It is the most important
differential stain used in
bacteriology because
it classified bacteria into
two major groups:

Gram positive: Gram negative:


Appears violet after Grams stain Appears red after Grams stain
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GRAM STAIN

Gram Stain:
Crystal violet (primary stain)
Grams iodine (mordant)
Acetone-alcohol (decolorizing agent)
Safranin (counter stain)

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Gram-positive bacteria
Have a thick peptidoglycan layer surrounds the cell.
The stain gets trapped into this layer and the bacteria
turned purple.
Retain the color of the primary stain (crystal violet) after
decolorization with alcohol

Gram-negative bacteria
have a thin peptidoglycan layer that does not retain crystal
violet stain.
Instead, it has a thick lipid layer which dissolved easily
upon decolorization with Aceton-Alcohol.
Therefore, cells will be counterstained with safranin and 13
turned red.
Crystal Violet 30-60 sec.
Rinse, Iodine 30-60 sec.
Rinse, Decolorize 5 sec or flash
Rinse, Saffranin 30-60 sec.
Rinse, blot dry
Cristal Violet

Iodine

Decolorization

Safranin
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Coccus
Staphylococcus / bergerombol seperti anggur
Gram positif
17
Contoh : Staphylococcus aureus
Coccus
Berantai / Streptococcus
Gram positif
18
Contoh : Streptococcus pyogenes
Basil
Streptobasil / Berantai
Contoh : Bacillus subtilis 19
Basil
Diplobasil
Contoh : Bacillus cereus 20
Coccus
Diplococcus
Gram negatif
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Contoh : Nesseria gonorhoea
Basil/batang
Gram negatif
Contoh : Escerichicia coli, Klebsiella sp 22
Koma/vibrio
Gram negatif
Contoh : Vibrio cholerae
23
Blastospora
Pseudohifa
Candida albicans 24