BATASAN HILLS NATIONAL HIGH SCHOOL

SENIOR HIGH SCHOOL - SCIENCE

GENERAL BIOLOGY

PREPARING ANIMAL AND PLANT CELL SLIDE

INTRODUCTION

There are many reasons to examine human cells and tissues under the microscope.
Medical and biological research is underpinned by knowledge of the normal structure and
function of cells and tissues and the organs and structures that they make up. In the normal
healthy state, the cells and other tissue elements are arranged in regular, recognizable
patterns. Changes induced by a wide range of chemical and physical influences are
reflected by alterations in the structure at a microscopic level, and many diseases are
characterized by typical structural and chemical abnormalities that differ from the normal
state. Identifying these changes and linking them to diseases is the basis of histopathology
and cytopathology, important specializations of modern medicine. Microscopy plays an
important part in haematology (the study of blood), microbiology (the study of
microorganisms including parasites and viruses), and more broadly in the areas of biology,
zoology, and botany. In all these disciplines, specimens are examined under a microscope.

There are many different forms of microscopy, but the one most employed is “brightfield”
microscopy where the specimen is illuminated with a beam of light that passes through it (as
opposed to a beam of electrons as in electron microscopy). The general requirements for a
specimen to be successfully examined using brightfield microscopy are:

 That the cells and other elements in the specimen are preserved in a “life-like” state
(this process is called “fixation”)
 That the specimen is transparent rather than opaque, so that light can pass through it
 That the specimen is thin and flat so that only a single layer of cells is present
 That some components have been differentially coloured (stained) so that they can
be clearly distinguished

OBJECTIVES

 To observe different types of plant and animal tissues.

 To prepare a slide of different type of tissues

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SENIOR HIGH SCHOOL - SCIENCE

PRE-LAB ACTIVITY

Direction: Observe the picture and label its parts.

EXPERIMENTATION
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SENIOR HIGH SCHOOL - SCIENCE

PART A: CROSS SECTION OF STEM TISSUE

Materials Needed:
Blade
Brush and Dropper Reagents Needed:
Needles Safranin
Watch Glass Balsam
Glass slide and cover slip
Compound microscope
herbaceous stem (Petchay/ Kangkong)

PROCEDURES:

 Take 2-3cm long pieces of the stem.

 Hold the material between thumb and first finger of your left hand.
 Hold the scalpel in the right hand with edge of the blade facing you and handle at
right angle to it.
 Dip the top of the material in water.
 Then start cutting transverse sections as fast as possible in a watch glass containing
water.
 Select the thinnest section of the material with the help of a delicate brush.
 Take a clean watch glass with water, transfer thin sections of the material.
 Put a few drops of saffranin stain in the watch glass with water.
 Leave it for 3-5 minutes.
 Drain off stain and wash with water if necessary.
 Put the thinnest section in the centre of the slide.
 Put a drop of balsam over the material.
 Cover it with a coverslip with the help of needle.
 Observe it under a compound microscope after staining and mounting.

PRECAUTIONS:
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SENIOR HIGH SCHOOL - SCIENCE

 Safranin is to be used to stain only the lignified tissues, over staining can be removed
by washing in water.
 Air bubbles must be avoided in the sections.
 Use only brush to transfer or to handle the sections. Do not use needles for this
purpose.
 Discard the incomplete and oblique sections.

OBSERVATION:

Draw and label the cross section of the stem using the High-power and low power
objectives.

HIGH POWER LOW POWER

Results and Discussion:

1. What is a vascular bundle? Describe its arrangement.

2. Describe the function of phloem and xylem.
3. How does a cuticle work?

Conclusion and Recommendation:

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SENIOR HIGH SCHOOL - SCIENCE

PART B: BONE TISSUE

Materials Needed:
Long bone of chicken
Hacks’ saw blade (Iron cutting)
Forceps
Sand paper Reagents Needed:
Tap water 50% alcohol
Glass slide and cover slip xylene
Compound microscope balsam
3 Petri dish

PROCEDURES:

 Two- to three-centimeter-thick transverse sections can be made by the hack's saw

blade.
 Thin sections thus taken from bone are later trimmed in appropriate size with the help
of scalpel and forceps thickness.
 The sandpaper placed first on a stable flat surface and the bone chip is placed on it.
With them palmar aspect of the tip of a finger (usually index or middle) the chip is
rubbed on the stone with horizontal to and fro movement maintaining a sustained
pressure. Water is applied frequently during the procedure with the painting brush.
Water acts as the lubricant-cum-coolant, which help in rapid progress of the work.
Repeated change of surface of the section with the forceps or paining brush helps in
uniformity of the both surface of the specimen. Fingers can be changed to avoid the
skin damage. Change of grinding pressure depends on the thickness of the section.
Achievement of transparency denotes the adequate thinning (nearly 50 micron) of
the specimen.
 Adequately thin specimen (nearly 50 micron) is now transferred to a Petridish
containg 50% alcohol for dehydration for 5 minutes. Dehydrated tissue is now
transferred to another Petridish containing xylene for 5 minutes. Xylene will make the
tissue further transparent.
 Check the specimen by transferring the tissue with the painting brush on a slide and
observing it under the low power magnification. If the specimen is good enough, next
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SENIOR HIGH SCHOOL - SCIENCE

step would be followed; otherwise further rubbing, dehydration and clearing steps
can be followed as necessary.
 Impregnation and mounting Specimen is mounted with thick balsam and cover slip
avoiding air-bubbles. After 24 hr drying it becomes ready to study under microscope

OBSERVATION:

Draw and label the parts of the specimen using the High-power and low power objectives.

HIGH POWER LOW POWER

Results and Discussion:

1. Describe one lamellae layer in bone.

2. What is in the center of every osteon?
3. Describe how a bone grows wider as you grow older and bigger.
4. Haversian Canals in bones must allow the blood to leave the bone once the
blood has given up its oxygen. Is this true? If so, why?(This will take some
thought involving all you’ve learned.)

Conclusion and Recommendation:

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SENIOR HIGH SCHOOL - SCIENCE

PART C Human Cheek Cell

Materials Needed:
Tooth pick
Dropper
Glass slide and cover slip Reagents Needed:
Filter paper xylene
Needle balsam
brush methylene blue solution

PROCEDURES:

 Gently scrape the inner side of the cheek using a toothpick, which will collect some
cheek cells.
 Place the cells on a glass slide that has water on it.
 Mix the water and the cheek cells using a needle and spread them.
 Take a few drops of Methylene blue solution using a dropper and add this to the
mixture on the slide.
 After 2-3 minutes remove any excess water and stain from the slide using a blotting
paper.
 Take a few drops of glycerine using a dropper and add this to the test mixture.
 Take a clean cover slip and lower it carefully on the mixture with the aid of a needle.
 Using a brush and needle, press the cover slip gently to spread the epithelial cells.
 Remove any extra liquid around the cover slip using a blotting paper.
 Place this glass side on the stage of the compound microscope and view it.

PRECAUTIONS:

 Ensure toothpick used to scrape the cheek is clean, so it does not cause infection to
the cheek.
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SENIOR HIGH SCHOOL - SCIENCE

OBSERVATION:

Draw and label the parts of the specimen using the High-power and low power objectives.

HIGH POWER LOW POWER

Results and Discussion:

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

2. Why do you think the cheek cells are a good a good source of cells for this
experiment?
3. Keeping in mind that the mouth is the first site of chemical digestion in a human.
Your saliva starts the process of breaking down the food you eat. Keeping this in
mind, what organelle do you think would be numerous inside the cells of your
mouth?

Conclusion and Recommendation:

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SENIOR HIGH SCHOOL - SCIENCE

References:

Department of Anatomy, North Bengal Medical College & Hospital, Sushrutanagar;

Darjeeling;
West Bengal 734 012.

https://biology4isc.weebly.com/slide-preparation.html

https://www.youtube.com/watch?v=WcYb-1h0hwc

https://amrita.olabs.edu.in/?sub=79&brch=15&sim=125&cnt=2