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Lab 3 Micro

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16 views13 pages

Lab 3 Micro

Uploaded by

tealempo80
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Al-Esraa University

Department of Pharmacy

By
Msc. Farah Hussein Ali

lab. 3
Smear preparation and
simple stain
Smear preparation
Principles : A bacterial smear is a dried preparation of
bacterial cell on class slide .
In bacterial smear that has been properly processed
1- the bacteria are evenly spread out on the slide in
such a concentration that they are adequately
separated from one anther
2- the bacteria are not washed off the slide during
staining
3- bacterial form is not distorted
Materials
1- 24 – 48 hours broth or agar of staphylococcus
aureus and bacillus subtilus

2- inoculation loop and needle

3-sterile distilled water


Procedure
Bacteria from either a broth culture or an agar
slant or plate may be used If slant or plate is
used a small amount of bacterial growth is
transferred to a drop of water on a glass slide
and mixed then the mixture is spread out evenly
over a large area on the slide If the medium is
liquid ,place one or two loops of the medium
directly on the slide and spread the bacteria
over large area .Allow the slide to air dry at
room temperature
The next step is to attach the bacteria to the
slide by heat fixing .this is accomplished by
gently heating ,passing the slide several time
through the hot portion of flame of a bunzen
burner . most bacteria can be fixed to the slide
and killed in this way without serious distortion
of cell structure .
Simple stain
Principles
The use of single stain or dye to create contract
between bacteria cell and back ground is
referred to as simple stain .some of the most
commonly used dyes for simple stain are crystal
violet ( 20-30 sec. ) ,methylene blue (60 sec .)
and carbol fuchsin (5 -10 sec.),All of these dyes
work well on bacteria because they have colore
–bearing ions that are positively charched
(cationic).
• The fact that are slightly negative charged
produces a pronounced attraction between
these cationic and the organism .Such dyes
are classified as basic dyes . The aim of using
these stain is to differentiate bacteria cells
from other cells on the slide and to recognize
the morphology and size and arrangement
bacteria
Materials

1- 24 – 48 hours broth or agar of


staphylococcus aureus and bacillus subtilus

2- Methylene blue
Procedure
1- Place the fixed smear on rack over a sink or
other suitable receptacle and stain the smear
with methylene blue for one minute
2-wash stain off slide with water for few seconds
3- dry the slide and examine under the oil
immersion
Thank you for listening

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