Cell and Molecular Biology BIO 171 Laboratory

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ACTIVITY NO. 2

CELL STAINING TECHNIQUES

Most types of cells do not have much natural pigment and are therefore difficult to see
under the light microscope unless they are stained. Several types of stains are used to make
bacterial cells more visible. Staining is a process in which cells or tissues are colorized with a
particular stain (chemical/dye) for identification. In addition, staining techniques can be used to
determine the cells’ biochemical or structural properties. This type of information can help
identify and classify the organism, and can be used by health care providers to diagnose the
cause of a bacterial infection.

Objectives:
This activity aims to distinguish the morphological shapes and arrangements of the cells
and different tissue types.

Questions:

1. Compare the two types of staining: simple staining and differential staining. (4 pts)
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2. Staining can be applied in-vivo or in-vitro. How will you differ each other? (4 pts)
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3. Staining can be classified based on its chemical nature: acidic stain, basic stain and neutral
stain. Differentiate each other. (6 pts)
Acidic stain

Basic stain

Neutral stain

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Cell and Molecular Biology BIO 171 Laboratory

4. Before specific staining can occur, tissue samples must undergo preparation through the
following steps below. Explain each step. (16 pts)
Fixation:

Dehydration:

Clearing:

Impregnation:

Embedding:

Sectioning:

Mounting:

Staining:

5. In simple staining, it can be classified into direct staining and indirect staining. Compare each
other based on the characteristics shown in the table below. (10 pts)

Characteristics Direct staining Indirect staining

Chemical nature of stain

Charge of stain

Chemical dye used

Appearance

Principle for discoloration

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Cell and Molecular Biology BIO 171 Laboratory

6. In differential staining, it can be categorized into acid-fast staining and gram staining. Explain
the resulting appearance on how it differs to one another. (16 pts)

Acid-fast staining: (definition)

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Resulting appearance:
Acid-fast bacteria Non-acid fast bacteria

Gram staining: (definition)

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Resulting appearance:
Gram-positive bacteria Gram-negative bacteria

7. The figure below demonstrates the basic concept of gram staining for a smear preparation.
Explain the purpose(s) of the chemicals or dyes used in the process. (8 pts)

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Cell and Molecular Biology BIO 171 Laboratory

Crystal violet:
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Iodine:
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Alcohol:
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Safranin:
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8. There are three basic bacterial shapes: coccus, bacillus, and spiral. In the table below, draw
the different shapes and arrangements of bacterial cells. (28 pts)
Coccus (spherical)

Cocci Diplococci Tetrad Sarcinae Streptococci Staphylococci

Bacillus (rod-shaped)

Bacilli Diplobacilli Streptobacilli Coccobacilli Palisades

Spiral (twisted)

Vibrio Spirillum Spirochete

9. Can a bacterial strain show both Gram's positive as well as Gram's negative reaction at the
same time? (3 pts)
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Cell and Molecular Biology BIO 171 Laboratory

10. In doing the staining process, problems may encounter on the resulting appearance.
Provide possible explanations to the specified errors on the table provided below. (10 pts)
Problems/Errors Explanations
Stain is too light or too dark.

No cells are found within the slide.

Cells look lysed or distorted.

Large crystals on the slide.

Slide not adhering. It appears that pieces

are floating.

11. There are variety of dye stains that identifies and demonstrates particular cell structures and
tissue types. Provide brief details on the given stains below. (15 pts)
Haematoxylin & Eosin stain

Orcein stain

Fuchsin stain

Wright’s Giemsa stain

Malachite green stain

References:

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