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Methylene Blue Staining Techniques

This document provides instructions for staining techniques used in a microbiology laboratory exercise. It describes procedures for simple staining using methylene blue to stain cheek cells and observe their structures under a light microscope. Key structures like the nucleus, cell membrane, and cytoplasm were visible, but organelles like the ER and Golgi apparatus were not. The document also outlines the steps for Gram staining, which can be used to differentiate between Gram-positive and Gram-negative bacteria.

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9 - MANLAPAZ, MARK EMIL B.
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233 views3 pages

Methylene Blue Staining Techniques

This document provides instructions for staining techniques used in a microbiology laboratory exercise. It describes procedures for simple staining using methylene blue to stain cheek cells and observe their structures under a light microscope. Key structures like the nucleus, cell membrane, and cytoplasm were visible, but organelles like the ER and Golgi apparatus were not. The document also outlines the steps for Gram staining, which can be used to differentiate between Gram-positive and Gram-negative bacteria.

Uploaded by

9 - MANLAPAZ, MARK EMIL B.
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd

Our Lady of Fatima University

College of Arts and Sciences


Natural Science Department
MICP 211
Laboratory Exercise
STAINING TECHNIQUE

SIMPLE STAINING
1.Procedure:

1. Put a drop of methylene blue on a slide. Caution: methylene blue will stain clothes and skin.
2. Gently scrape the inside of your cheek with the flat side of a toothpick/cotton swab. Scrape
lightly.
3. Stir the end of the toothpick/cotton swab in the stain and throw the toothpick away.
4. Place a coverslip onto the slide
5. Use the SCANNING objective to focus. You probably will not see the cells at this power.
6. Switch to low power. Cells should be visible, but they will be small and look like nearly clear
purplish blobs. If you are looking at something very dark purple, it is probably not a cell
7. Once you think you have located a cell, switch to high power and refocus. (Remember, do NOT
use the coarse adjustment knob at this point)

2. Describe or define each of the following

 Cell membrane - A semi permeable layer surrounding the cytoplasm and outside of the
cell. Allows certain things in and out.
 Cytoplasm - the material in the cell surround the nucleus and organelles.
 Nucleus - Controls cell activity, and stores DNA.
 Organelle - a "tiny organ", part of a cell where cellular activities take place

3. Sketch the cell at low and high power. Label the nucleus, cytoplasm, and cell membrane. Draw your
cells to scale.
4. Why is methylene blue necessary?

- It is a stain, so it stains the cells in order for us to clearly see them and differentiate them.

5. The light microscope used in the lab is not powerful enough to view other organelles in the
cheek cell. What parts of the cell were visible?

-The parts visible were the nucleus, cytoplasm, and the cell membrane.

6. List 2 organelles that were NOT visible but should have been in the cheek cell.

- They should have had the Smooth and Rough ER and the Golgi Apparatus.

7. Is the cheek cell a eukaryote or prokaryote? How do you know?


- It is a eukaryote because it has a nucleus and a membrane.

GRAM STAINING TECHNIQUE:

1. Prepare smear on a clean slide

2. Stain with crystal violet for 30 seconds.

3. Rinse with water

4. Flood the film with Grams iodine and allow it to act for 30 sec.

5. Rinse with water.

6. Decolorized with 95% alcohol.

7. Rinse with water.

8. Counter stain with safranin for 20-30 sec.

9. Rinse with water and blot dry.

10. Examine under oil immersion objective

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