Location via proxy:   [ UP ]  
[Report a bug]   [Manage cookies]                
Scribd
Upload
30 views

Micro Lab

This document outlines the steps of the Gram stain procedure, which is used to categorize and identify bacterial strains based on differences in their cell wall composition. Gram-positive bacteria appear dark purple due to retaining the initial violet stain in their thick peptidoglycan layer, while Gram-negative bacteria appear pink from the counterstain as their thinner peptidoglycan allows the violet stain to be washed away. The 15-step procedure is described in detail. Sample results showed unclear vision, indicating errors were made such as using excessive pigments or uneven distribution on the slide. Protection and prevention for students working with microbes is suggested.

Uploaded by

Nser Seyam
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
30 views

Micro Lab

This document outlines the steps of the Gram stain procedure, which is used to categorize and identify bacterial strains based on differences in their cell wall composition. Gram-positive bacteria appear dark purple due to retaining the initial violet stain in their thick peptidoglycan layer, while Gram-negative bacteria appear pink from the counterstain as their thinner peptidoglycan allows the violet stain to be washed away. The 15-step procedure is described in detail. Sample results showed unclear vision, indicating errors were made such as using excessive pigments or uneven distribution on the slide. Protection and prevention for students working with microbes is suggested.

Uploaded by

Nser Seyam
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
You are on page 1/ 6

College of Health Professions

Human Medicine
Medical Microbiology Laboratory) (19057
Gram stain procedure

Name: Jasem Yousef Hroub


ID:200087
DR.Odai Rjoub
Introduction:
Gram staining, named for it’s originator in 1884, is the central
method in microbiology for categrorizing and identifying bacterial
strains. The traditional five-steps process results in two differently-
colored fixed bacterial cells, depending on their cell wall
composition. Gram-positive will appear dark purple (from the initial
violet stain) and gram-negative will be pink to red color from the
final counterstain of safranin.
The different colors are due to a polymer called peptidoglycan that
is a part of the bacterial cell wall. Gram-positive bacteria have a
majority (50-90%) of peptidoglycan present, and Gram-negative
bacteria have a much smaller proportion (often only 10%). This
affects the amount of violet dye that is retained during the process.
Since the Gram- negative bacteria have so little dye to start with, it is
quickly removed during the procedure, and the final safranin
counterstain adds the pink color to aid visibility.
Aims:
● To distinguish between different types of bacteria using gram
stain.
● To classify bacteria as gram-positive and gram-negative
bacteria.

Materials:
● Gram stain(Crystal violet (primary stain) Iodine solution/Gram's
Iodine (mordant that fixes crystal violet to cell wall) Decolorizer
(e.g. ethanol) Safranin (secondary stain) Water (preferably in a
squirt bottle)
● Microscope slides
● Heat source
● Sterile swab
● Culture or specimen
● Microscope
methods:
1. Compile all necessary tools and supplies.
2. Add the patient's initials or the specimen number to the slide's
label.
3. Select an isolated colony from the agar plate, then collect bacteria
with a clean swab.
4. Spread the colonies out in a circular pattern on the microscope
slide after placing the swab there.
5. Heat fix the microorganisms to the slide by Pass the slide through
the flame several times to fix the smear (30 seconds.)
6. Place slide on staining tray or hold with forceps above the sink.
7. Crystal Violet stain should be applied liberally and left to settle on
the slide's surface for a minute.
8. Use distilled water to rinse the slide.
9. Pour Gram's Iodine all over the slide and let it sit for a minute.
10. Use distilled water to rinse the slide.
11. Flood the slide with Gram’s decolorizer and time for 30
seconds.
12. Use distilled water to rinse the slide.
13. Flood the slide with the counterstain, Safranin, and let sit for
one minute.
14. Use distilled water to rinse the slide.
15. Blot the slide before reading with the microscope's oil
immersion lens. Gram-negative and Gram-positive bacteria should
be sought out.
Result:

Sample 1: Sample 2:

Discussion:
The vision of the samples was not very clear after the completion of
the practical side, and this indicates that there are a number of
errors that occurred, among these errors is the excessive amount of
different pigments, and the distribution of pigments to all sides of
the slide.
Suggestion:
Providing appropriate means of protection and prevention for
students.
Enhancing students’ awareness of the dangers of dealing with
different types of microbes.

Reference:
-AAT Bioquest, What is Gram staining ?

You might also like