MABALACAT CITY COLLEGE

INSTITUTE OF ARTS, SCIENCES AND TEACHER

EDUCATION
(IASTE)

MODULE 4
(November 30 – December 05, 2020)

BSSE 119
BIOTECHNIQUES

ROWENA V. ALDANA
Instructor

MABALACAT CITY COLLEGE

INSTITUTE OF ARTS, SCIENCES AND TEACHER
EDUCATION
 Mabalacat City, Pampanga

Module 4 – BSSE 119 - BIOTECHNIQUES

SLIDE MAKING TECHNIQUES
Compiled and Prepared by ROWENA V. ALDANA
1st Semester A.Y. 2020-2021

I. OVERVIEW

This module deals with the understanding and discusses different procedures on how to prepare a
wet mount and dry mount slide in viewing plant cells and animal cells.

II. LEARNING OBJECTIVES

This module is designed for you to:

• Prepare a wet mount and dry mount slide for use with a microscope.
• Use a microscope correctly and safely to find and observe components of plant cells
and animal cells

III. How to complete this module?

1. Read the given module

2. Complete the ACTIVITY and EVALUATION. Due date of your accomplished worksheets will be on
December 12, 2020

3. If any questions or clarification on the lesson, leave a message in our Facebook group chat or in my Facebook
messenger https://www.facebook.com/Rowena V. Aldana or email me at [email protected], from
Saturday at 8 AM to 11 AM.

4. You can use the following link provided for additional video presentations that you can use as references.
https://www.youtube.com/watch?v=R8fos4zWSII
https://www.youtube.com/watch?v=4Nurc1hSN4o&t=60s

IV. LECTURE

 
Microscope slides are pieces of transparent glass or plastic that support a sample so that they can
be viewed using a light microscope. There are different types of microscopes and also different
types of samples, so there is more than one way to prepare a microscope slide. The method used
to prepare a slide depends on the nature of the specimen. Three of the most common methods are
wet mounts, dry mounts, and smears.

Wet Mount Slides

Wet mounts are used for living samples, transparent liquids, and
aquatic samples. A wet mount is like a sandwich. The bottom layer
is the slide. Next is the liquid sample. A small square of clear glass
or plastic (a coverslip) is placed on top of the liquid to minimize
evaporation and protect the microscope lens from exposure to the
sample.

To prepare a wet mount using a flat slide or a depression slide:

1. Place a drop of fluid in the middle of the slide (e.g., water, glycerin, immersion oil, or a
liquid sample).
2. If viewing a sample not already in the liquid, use tweezers to position the specimen
within the drop.
3. Place one side of a coverslip at an angle so that its edge touches the slide and the outer
edge of the drop.
4. Slowly lower the coverslip, avoiding air bubbles. Most problems with air bubbles come
from not applying the coverslip at an angle, not touching the liquid drop, or from using
a viscous (thick) liquid. If the liquid drop is too large, the coverslip will float on the slide,
making it hard to focus on the subject using a microscope.

Some living organisms move too quickly to be observed in a wet mount. One solution is to add a
drop of a commercial preparation called "Proto Slow." A drop of the solution is added to the
liquid drop before applying the coverslip.

Some organisms (like Paramecium) need more space than what forms between a coverslip and a
flat slide. Adding a couple of strands of cotton from tissue or swab or else adding tiny bits of
broken coverslip will add space and "corral" the organisms.

As the liquid evaporates from the edges of the slide, living samples may die. One way to retard
evaporation is to use a toothpick to coat the edges of the coverslip with a thin rim of petroleum
jelly before dropping the coverslip over the sample. Press gently on the coverslip to remove air
bubbles and seal the slide.

Dry Mount Slides

Dry mount slides can consist of a sample placed on a slide or
else a sample covered with a coverslip. For a low power
microscope, such as a dissection scope, the size of the object
isn't critical, since its surface will be examined. For a
compound microscope, the sample needs to be very thin and
as flat as possible. Aim for one cell thickness to a few cells.
It may be necessary to use a knife or razor blade to shave a
section of the sample.

1. Place the slide on a flat surface.

2. Use tweezers or a forceps to place the sample on the slide.
3. Place the coverslip on top of the sample. In some cases, it's okay to view the sample
without a coverslip, as long as care is taken not to bump the sample into the microscope
lens. If the sample is soft, a "squash slide" may be made by gently pressing down on the
coverslip.

If the sample won't stay on the slide, it may be secured by painting the slide with clear nail polish
immediately before adding the specimen. This also makes the slide semipermanent. Usually,
slides can be rinsed and reused, but using nail polish means the slides must be cleaned with
polish remover before reuse.

How to Make a Blood Smear Slide

Some liquids are either too deeply colored or too thick to view
using the wet mount technique. Blood and semen are prepared
as smears. Evenly smearing the sample across the slide makes it
possible to distinguish individual cells. While making a smear
isn't complicated, getting an even layer takes practice.

1. Place a small drop of a liquid sample onto the slide.

2. Take a second clean slide. Hold it at an angle to the first slide. Use the edge of this slide
to touch the drop. Capillary action will draw the liquid into a line where the flat edge of
the second slide touches the first slide. Evenly draw the second slide across the surface of
the first slide, creating a smear. It's not necessary to apply pressure.
3. At this point, either allow the slide to dry so that it can be stained or else place a coverslip
on top of the smear.

How to Stain Slides

There are many methods of staining slides. Stains make it

easier to see details that might otherwise be invisible.
Simple stains include iodine, crystal violet, or methylene blue. These solutions may be used to
increase contrast in wet or dry mounts. To use one of these stains:

1. Prepare a wet mount or dry mount with a coverslip.

2. Add a small drop of stain to an edge of the coverslip.
3. Place the edge of a tissue or paper towel on the opposite edge of the coverslip. Capillary
action will pull the dye across the slide to stain the specimen.

Common Objects to Examine with a Microscope

Many common foods and objects make fascinating subjects for slides. Wet mount slides are best
for food. Dry mount slides are good for dry chemicals. Examples of appropriate subjects include:

 Table salt
 Epsom Salt
 Alum
 Dishwashing detergent powder
 Sugar
 Mold from bread or fruit
 Thin slices of fruits or vegetables
 Human or pet hair
 Pond water
 Garden soil (as a wet mount)
 Yogurt
 Dust

V. EVALUATION:

MABALACAT CITY COLLEGE

INSTITUTE OF ARTS, SCIENCES AND TEACHER
EDUCATION
Mabalacat City, Pampanga

Module 4 – BSSE 119 - BIOTECHNIQUES

SLIDE MAKING TECHNIQUES
Compiled and Prepared by Rowena V. Aldana 1st Semester A.Y. 2020-2021

Name: _______________________________ Score:____________________

Course/Section:____________________________ Date: ____________________

ACTIVITY 1: Encircle the letter of the best answer

1. Which type of slides are the easiest to prepare?

a. dry mount c. wet mount
 b. prepared mount d. electron
2. Which of the following is one advantage of putting liquid on a specimen, as used in wet mount
slides?
a. More permanent slide fixture
b. Magnification of the specimen
c. Flattens the specimen
d. Kills the specimen
3. What must be added to a prepared mount slide to permanently preserve and adhere the
specimen to the slide and cover slip?
a. Stains b. A fixative c. Mordant d. Mounting medium
4. The object to be studied and placed on the stage of the microscope is described as the ____
a. slide b. image c. clip d. specimen
5. A human cheek cell is best described as an example of ____
a. a protist c. a plant cell
b. an animal cell d. an organism
6. In preparing temporary mounts for microscopic examination, why the specimen has to be thin?
a. To make the specimen easier to be focused
b. To make the specimen easier to be covered by the cover slip
c. To reduce the amount of stain used
d. To allow light pass through the specimen to the objective
7. In preparing a slide, why the specimen is stained?
a. To make the cells bigger
b. To separate the cells
c. To make the specimen more colorful
d. To make the structures easier to be seen
8. The following diagram shows the technique in lowering the cover slip to cover the specimen.
Why must the edge of the cover slip touch the stain before it is lowered?

a. To make sure the stain spread out evenly.

b. To protect the specimen from breaking into pieces.
c. To keep the temporary mount tidy for observation under a microscope.
d. To avoid the formation of air bubbles under the cover slip.
9. Which of the following statements about observation under a microscope is correct?
a. Under low power magnification, the area of specimen observed is
larger.
b. Under low power magnification, a more detailed image is observed.
c. Under low power magnification, the image is darker.
d. Under high power magnification, the image is lighter.
10. It is difficult to see the nuclei of cells under light microscope. Which of the following would
help to make the nuclei more visible?
a. staining the cells before mounting.
 b. using a thinner glass slide for mounting the cells.
c. increasing the illumination of the cells.
d. reducing the distance between the cells and the objective lens.

VI. REFERENCES:

https://projects.ncsu.edu/cals/course/pp318/profiles_mirror/pm/techniques.html
https://www.thoughtco.com/how-to-prepare-microscope-slides-4151127

Disclaimer:
It is not the intention of the author/s nor the publisher of this module to have monetary gain in
using the textual information, imageries, and other references used in its production. This module is only
for the exclusive use of a bona fide student of Mabalacat City College.
 In addition, this module or no part of it thereof may be reproduced, stored in a retrieval system,
or transmitted, in any form or by any means, electronic, mechanical, photocopying, and/or otherwise,
without the prior permission of Mabalacat City College.