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Microbiology Laboratory Lab Report # 6: Simple Staining Name: Pantoja, Ariel S. Date: - Year and Section: 1-Y2-12

Simple staining uses a single basic stain like methylene blue or crystal violet to elucidate bacterial cell morphology and arrangement by producing contrast between organisms and their background, as basic stains are attracted to negatively charged cell components; while it cannot differentiate Gram positive and negative bacteria, its purpose is to visualize rod-shaped, spiral, and spherical cell shapes in purple. The principle of simple staining relies on basic dyes containing positively charged chromophores strongly binding to negatively charged structures like nucleic acids and cell walls.

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81 views2 pages

Microbiology Laboratory Lab Report # 6: Simple Staining Name: Pantoja, Ariel S. Date: - Year and Section: 1-Y2-12

Simple staining uses a single basic stain like methylene blue or crystal violet to elucidate bacterial cell morphology and arrangement by producing contrast between organisms and their background, as basic stains are attracted to negatively charged cell components; while it cannot differentiate Gram positive and negative bacteria, its purpose is to visualize rod-shaped, spiral, and spherical cell shapes in purple. The principle of simple staining relies on basic dyes containing positively charged chromophores strongly binding to negatively charged structures like nucleic acids and cell walls.

Uploaded by

Christopher Galivo
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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MICROBIOLOGY LABORATORY

Lab Report # 6:
SIMPLE STAINING
NAME: PANTOJA, ARIEL S. Date: ____________
Year and Section: 1-Y2-12

I. Research from open educational resources and attach a diagram of simple staining
procedure. ( Please cite your references )or perform the simple staining procedure
using do it yourself materials at home and attach.

INTRODUCTION
In simple staining, the bacterial smear is stained with a single reagent, which produces
a distinctive contrast between the organism and its background. Basic stains with a
positively charged chromogen are preferred because bacterial
nucleic acids and certain cell wall components carry a negative charge that strongly
attracts and binds to the cationic chromogen. The purpose of simple staining is to
elucidate the morphology and arrangement of bacterial cells. The most commonly used
basic stains are methylene blue, crystal violet, and carbol fuchsin.

PROCEDURE
1. Place a slide on the staining tray and flood the smear with one of the indicated
stains, using the appropriate exposure time for each: carbol fuchsin, 15 to 30
seconds; crystal violet, 20 to 60 seconds; methylene blue, 1 to 2 minutes.
2. Gently wash the smear with tap water to remove excess stain. During this step,
hold the slide parallel to the stream of water; in this way you can reduce the loss
of organisms from the preparation.
3. Using bibulous paper, blot dry, but do not wipe the slide.
4. Examine all stained slides under oil immersion.

INTERPRETATION
Bacilli and diplobacilli: Rod-shaped bacteria, purple
Spirilla: spiral-shaped bacteria, purple
Cocci: spherical-shaped, bacteria, purple

REFERENCE
https://www.youtube.com/watch?v=kaOJDG_PnDQ
I. Look for photomicrographs of different kinds, shapes and arrangements of
bacteria ( diplococci, streptococci, staphylococci, bacilli, vibrio etc. ) as seen
under the microscope using simple staining.

II. Answer the following:


1. What is the importance of simple staining?
- The purpose of simple staining is to elucidate the morphology and
arrangement of bacterial cells. The most commonly used basic stains are
methylene blue, crystal violet, and carbol fuchsin.
2. Can you differentiate Gram positive and Gram negative bacteria using a simple
staining procedure?Why?
- The Gram stain procedure distinguishes between Gram positive and Gram
negative groups by coloring these cells red or violet. Gram positive
bacteria stain violet due to the presence of a thick layer of peptidoglycan
in their cell walls, which retains the crystal violet these cells are stained
with.

3. Explain the principle behind simple staining


- Its principle is based on producing a marked contrast between the
organism and its surroundings by using basic stain. A basic dye consists
of a positive chromophore, which strongly attracts to the negative cell
components and charged molecules like nucleic acids and proteins.

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