Biology Lab Manual 2024 - 25

ST. PATRICK SENIOR SECONDARY SCHOOL

BIOLOGY PRACTICAL MANUAL
2024-2025
___________________________________________________________________________________________________

INSTRUCTIONS FOR WRITING RECORD :

 Use only blue pen

 Write theory in ruled pages

 Diagrams and tabular forms in unruled pages.

 Diagrams and margins of tabular forms only with pencil.

 Every experiment start in a fresh page.

 Don’t write index page and dates of experiments.

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 Biology Lab Manual 2024 - 25
SECTION - A
EXP:1 PREPARATION OF
F TEMPORARY MOUNT TO OBSERVE POLLEN
GERMINTAION
AIM:
To study and calculate the percentage of pollen germination on a slide.

Pollen Grain Germination: In nature, pollen grains germinate on the compatible stigma of the carpel.
This process is called pollen-pistil interaction. It is known to be an essential step in fertilisation of
angiosperms which determines the compatibility and incompatibility of both pollen and pistil. This
dynamic process involves pollen recognition followed by inhibition or promotion of pollen
germination. The pollen grains can be induced to germinate in a synthetic medium using chemicals
which favour pollen germination such as 10% sucrose solution that facilitates rapid growth of pollen
tubes. During germination, intine (inner wall) of pollen grain emerges out as pollen tube through one
of the germ pores in exine (outer wall) and the pollen tube grows into the genetically compatible
stigma only.
Out of two male gametes carried in the pollen tube, one fuses with the egg cell while, the other ruses
with the secondary nucleus. This process is known as double fertilisation, which is a characteristic
feature of all angiosperms. The pollen of different flowers might differ in their germination time and
length of the pollen tubes. They also show different sculpturing on their pollen wall.

REQUIREMENTS:
Mature pollens of any seasonal flower like Trade scantia, jasmine, lily, etc., cavity glass slides,
cover slips, microscope, distilled water, beaker, weighing machine, filter paper, glass rod, boric acid,
sucrose, calcium nitrate, brushes and dropper.

PROCEDURE:
i. Firstly, prepare a pollen germination medium by dissolving 10 g sucrose,30 mg calcium nitrate
and 10 mg boric acid in 100 mL of distilled water. Alternatively, 10% sucrose solution can also
be used as the medium.
ii. Put a drop of this prepared medium or 10% sucrose solution with the help of a dropper in the
cavity slide.
iii. Dust the pollen on the medium on same slide with the help of a drawing brush.
iv. Now, cover pollens with the cover slip.

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 Biology Lab Manual 2024 - 25
v. Observe the slide containing pollen under the microscope after 10-15 minutes:
vi. Finally, count the total number of nor germinating pollen and germinating pollen grains

OBSERVATION:
Count the germinated and total no. of pollens per microscopic field and tabulate your readings in the
following table
% of pollen
Name of the No. of pollens showing Total no. of germination
S.No a
Flower germination (a) pollens seen (b) 100
b
1 Hibiscus 10 20
10
100 = 50%
20

CALCULATION:
a
Percentage of pollen germination = 100 or 100a
b b
where, a = Number of pollens showing germination, b = Total number of pollens

RESULT:
All the pollens of different species in the medium germinate with different percentage. Also, there is
great variation in time taken for the germination of pollen of each species.

PRECAUTIONS:
• The nutrient solution should be made carefully otherwise, pollen grains might fail to germinate.
• Only few pollen grains should be dusted in the nutrient solution in order to avoid the
overloading or overcrowding.
• While placing cover slip, air bubbles should be avoided else they might interfere with
observation and counting of pollens.
• The entire process should be carried at room temperature.
• Isolation of DNA from available plant material like Pea seed, Spinach, Onion, etc

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 Biology Lab Manual 2024 - 25
EXP : 2 TO STUDY THE PLANT POPULATION DENSITY BY QUADRAT METHOD
AIM:
To study the plant population density by quadrat method.

PRINCIPLE:
Population Density:
It represents the numerical strength of a particular plant species in the community per unit area at a
specific time. The unit area may be small or large, depending on the size and nature of the plant
community being studied.
Population density of an organism in an ecosystem varies from place to place and depends on various
factors such as food and water availability, predation, weather conditions, birth rate, death rate and
migration rates. It can be calculated using the following formula:
Total no. of individuals of the species in all the sampling units ( S)
Density (D)=
Total no. of sampling units Studied ( Q)

Total number of individual(s) of the species in all the sampling units (S)
Total number of sampling units studied (Q)
The value thus obtained is then expressed as number of individuals per unit area.

Quadrat Method:
Quadrats are the sampling units of a known area. These sampling units must be distinct and must not
overlap each other. The quadrats having a rectangular frame and dimensions of 1m x1m are most
commonly used for herbaceous plants. Quadrat method is considered as the most preferable method
to calculate population density and percentage frequency of different plant species

REQUIRMENTS:
Cotton/nylon thread (five meters), nails, a hammer or wooden quadrat of size 1mx 1m, paper, pencil,
hand lens, etc.

PROCEDURE:
i. Select an area having varied vegetation for study.
ii. Fix four nails with the help of hammer in the selected area such that the area enclosed by them
is a square of 1m X1m dimension. Make the boundaries of the square by tieing thread over the
nails.
Or
You can also use a commercially available ready-made quadrat of size 1m x1m. Keep it over
thearea under study.
iii. Note down the names of all the plant species seen in the quadrat. If the names of the plants are
not known, assign alphabets such as A, B, C, D, etc. to them.
iv. Count the number of individuals of a plant species present in the quadrat and note down the
number in front of the species name.
v. Repeat the same step (4) for every plant species present in the quadrat. Record the data as
shown in the observation table ahead.

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 Biology Lab Manual 2024 - 25
vi. Now, repeat the steps 1 to 5 to obtain data of nine more quadrats placed randomly at the site
of study. Take care that all the regions taken as sample areas should be distinct and non-
overlapping.

OBSERVATIONS:
Write down the total number of species seen in ten quadrats. This will give an idea about the
composition of the vegetation. There will be difference in the species composition in the quadrats
made in shady areas, exposed areas with bright sunlight, dry or wet areas, etc.

RESULT:
The population density of a plant species in a given field can be calculated by the formula given below:
S
D=
Q
PRECAUTIONS:
• The measurement of the quadrats should be accurate
• Count the individuals of one plant species at a time.
• Squares should be marked from one field only and they should not be very far from each other.
• Make sure that the vegetation is not damaged while laying the quadrat.

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 Biology Lab Manual 2024 - 25
EXP: 3 STUDY PLANT POPULATION FREQUENCY BY QUADRAT METHOD

AIM:
To study plant population frequency by quadrat method.

PRINCIPLE:
Population is known as the aggregation of individuals of the same kind inhabitating a particular place
or a particular geographical area at a particular time. Frequency is concerned with the degree of
uniformity of the occurrence of individuals of a species within a plant community. It is measured by
noting the presence of a species in random sample areas (quadrats) which are distributed as widely as
possible throughout the area of study. Frequency can be determined by means of sample areas in
order to check general impressions about relative values of species. Frequency is the number of
sampling units (as%) in which a particular species (A) occurs. The frequency of each species (sps. A or
sps. B or sps. X, etc) is expressed in percentage and is calculated as follows:

% Frequencv or Frequencv index (F)

Number of sampling units (quadrats) in which the species occurs (N)
Total number of sampling units (quadrats) employed for the study (Q)

REQUIREMENTS:
Cotton/nylon thread of 5 meters, nails, hammer, paper, pencil and magnifying lens.

PROCEDURE:
i. Select a field and lay a quadrat with the help of meter scale.
ii. Measure area of about 1mx 1m with the meter scale. Hammer the nails firmly at corners and
tie the thread on the nails.
iii. Divide each quadrat into 16 small squares by tying thread at a distance of 25 cm each on either
side, fix nails to the thread to mark these small squares.
iv. List the names of the plant species seen in the quadrat (if the name is not known mark these as
species A or B, etc. and if the same species is seen in other quadrats assign the same alphabet).
v. Similarly, lay ten more quadrats randomly in the field of study and record the number of
individuals of each species.
vi. Calculate the percentage frequency of occurrence of plants using the formula given below

Frequency Percentage for a species = No.of quadrants in which species appeared(N)

Total number of quadrants studied(Q)
x100
where, F= Frequency or frequency index
N = Number of sampling units (quadrats) in which the species occurs
Q = Total number of (quadrats) employed for the study

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 Biology Lab Manual 2024 - 25

OBSERVATIONS
Record the total number of species seen in the eleven quadrats. This will give an idea about the
composition of the vegetation. There will be a difference in the species composition in the quadrats
made in shady areas, exposed areas with bright sunlight, dry or wet areas, etc. Observe that the
frequency of occurrence is not the same for all species.
Percentag
Number of
e
quadrats in
Plant Frequenc
Quadrats employed in the study which the
Species y
species is
F=
present (N)
N/Q*100
I II III IV V VI VII VIII IX X
X P P P P P P P 7 70%
Y P P 2 20%
Z P 1 10%

RESULT:
The frequency percentage of a given species F = N 100
Q

PRECAUTIONS:
• The measurement of quadrats should be accurate.
• The string or cord used should not be very thick.
• One individual of a species should be counted only once in the quadrat.
• Make sure that the vegetation is not damaged, while laying the quadrat.

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 Biology Lab Manual 2024 - 25
EXP: 4 PREPARATION OF TEMPORARY MOUNT OF ONION ROOT TIPS TO STUDY
MITOSIS.
AIM:
To prepare a temporary mount of onion root tips to study mitosis.

Requirements:
Onion bulbs, wide mouthed glass bottle, corked tube, petri dishes, scissors, forceps, needles, methyl
alcohol, acetic acid, N/10 HCL, acetocarmine, distilled water, spirit lamp, compound microscope, slides,
blotting paper, cover slips etc.

Procedure:
A. Growing Onion root tips:
i. Select medium onion bulbs and carefully move remove the dry roots attached to them.
ii. Place them in a bottle filled with water in such a way that only roots touch the water level.
iii. Check the water level in the bottle everyday and add few drops water periodically in order to
compensate any evaporation losses.
iv. Keep them in position for about 3-6days. New roots may take 3- 6days to grow.
v. Cut 2-3cm long freshly grown roots, 2hrs after the sun rice and keep harvested roots in fixative
(Glacial acetic acid and ethanol in 1:3 ratio) for 2hours.
vi. Transfer roots from fixative to 70% alcohol.
vii. These root tips are ideal material for the study of mitotic cell division.

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 Biology Lab Manual 2024 - 25
B. Preparation and study of slide
i. Take one or two preserved roots and wash them thoroughly in water on a clean slide.
ii. Place one drop of N/10 HCL on the root tip followed by 2-3 drops of acetocarmine stain on it.
Leave it for 5-10minutes
iii. Slightly warm the slide on spirit lamp. Care should be taken that the stain is not dried up.
iv. Blot the excess stain using blotting paper.
v. Cut the more stained tip portion of root and retain it on slide and discard the remaining portion.
vi. After 10-20 seconds, put one or two drops of water and blot them carefully using blotting paper.
vii. Gently squash the root by tapping the cover slip with the blunt end of a needle so that
meristematic tissue of that root tip and is smashed and spread as a layer.
viii. There should no air bubbles under cover slip.
ix. Gently warm the slide over a flame and seal the margins of cover slip using paraffin wax.
x. Slide is now ready for the study of mitosis.
xi. Observe it in a compound microscope.

OBSERVATIONS:
Under lower magnification of the microscope, rectangular cells with pink nucleus are seen scattered.
Under higher magnification of the microscope different phases of mitosis become distinct. The stages
of mitosis can be broadly categorised into two main parts, i.e., karyokinesis followed by cytokinesis.
Those cells which are not in the phases of cell division are considered to be in the interphase. You will
observe that most cells in a microscopic field are in interphase.
Interphase

i. It is a non-dividing phase of the cell cycle occurring betweentwo successive cell divisions.
ii. The cells are mostly rectangular, oval or even circular in shape.

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 iii. The nucleus is homogenous and looks granular in origin.
iv. Nuclear envelope is distinct,
v. Chromatin fibres appear in the form of an interconnectedmass within the nucleus.
vi. Nucleolus is well observed inside the nucleus.

Stages of Mitosis
a) Prophase
i. Nucleus is enlarged and occupies most of the cell volume. Intact nuclear outline is seen.
ii. Chromatin network gets condensed and appears as long thread-like structures called
chromosomes.
iii. Nuclear membrane may start disappearing.
iv. Nucleoli may or may not be visible.
v. If the cell is in early stage of prophase, then the chromatin fibres are very thin. However, in the
cells at late prophase, comparatively thicker chromatin fibres would be visible it.

(a) Early prophase (b) Late prophase

b) Metaphase

i. Chromosomes become shorter and thicker and hence become distinct and clearly visible under
the compound microscope.
ii. Nuclear membrane completely disappears.
iii. Chromosomes orient themselves towards the equator with their centromeres arranged on an
equatorial line forming metaphase plate. Out of two chromatids of each chromosome, one
chromatid faces one pole and the other chromatid faces the opposite pole.
iv. Series of spindle fibres attach the centromeres to the opposite poles.
v. Nucleolus is not observed during metaphase.

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 c) Anaphase

(a) Early Anaphase (b) Late Anaphase

i. The two sister chromatids of each chromosome separate due to the splitting of centromere
and move towards the opposite poles.
ii. The daughter chromosomes (separated chromatids) appear in V, J, L and I shapes, depending
upon the position of the centromere in the chromosomes.
iii. Anaphase is designated as early, mid and late stages depending upon the position of moving
chromosomes toward the opposite poles.
d) Telophase

Chromosomes reach to the

opposite poles, lose their individuality and look like a mass of chromatin again.
i. Nuclear membrane reappears to form the nuclei of two future daughter cells.
ii. Nucleolus gets reconstituted.
iii. Two daughter nuclei are thus formed and appear to be similar to the parent nucleus both
quantitatively and qualitatively.
Cytokinesis

(A) Animal cells (B) Plant cells

In plant cells, cell plate is formed in the middle after telophase. The plate can be seen to extend
outward to ultimately reach the margin of the cell and divide the cell into two equal halves. Such cell

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plates are characteristic of plant cells. In animal cell, furrow appears in the membrane at both sides
and grows inward to divide cells into two.

RESULT:
All the stages of mitotic cell division are clearly visible in the slide prepared from onion root tips.

PRECAUTIONS:
i. Always clean the slide and cover slip thoroughly before and after use.
ii. Care should be taken that there should be no air bubbles under the cover slip
iii. The base of the onion bulb should exactly be in contact with water, while growing the roots.
iv. Always filter the acetocarmine stain before use.
v. Material should be mounted on the centre of slide.
vi. Also, the material should only be warmed gently, i.e. do not allow solution to boil on slide while
warming.

EXP: 5 ISOLATION OF DNA FROM AVIALBLE PLANT MATERIAL

AIM:
To isolate DNA from available plant material like pea seed, spinach, onion, etc.

REQUIREMENTS:
Plant material (like pea-seed, spinach leaves, onion or cauliflower shavings), mortar and pestle,
beakers, test tubes, enzymes (like cellulase, protease, lipase and ribonuclease), ethanol, spool, glass
rod, etc.

PROCEDURE:
i. Take small amount of plant material (onion or cauliflower shavings) and grind it in a mortar
with the help of pestle. Alternatively, the plant material can be homogenised in a blender and
can be filtered easily through the muslin cloth.
ii. Now, pour the filtrate into a boiling test tube.
iii. Break the cell wall and envelope of plant cell by treating the material with the enzyme
cellulase.
iv. Now, the same material is treated with enzyme protease to remove histone proteins.
v. Now, add ribonuclease in it to dissolve the associated RNA followed by addition of lipase to
dissolve the lipids present in the sample.
vi. Gradually pour twice the volume of ice-cold 95% ethanol into the test tube. This will allow DNA
to precipitate out.
vii. The precipitated DNA can be spooled with the help of glass rod. It appears as the winding of the
fine threads of DNA.

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OBSERVATIONS:
Shiny white DNA can be seen at the junction of solution and ethanol. This DNA represents all the DNA
found in plant cells. The chromosomes were broken in the process and the DNA gets precipitated due
to the chemical treatment.

RESULT:
DNA appears as white precipitate of very fine threads on the spool.

PRECAUTIONS:
• Fresh plant materials should be used for extraction of DNA.
• The glass wares must be thoroughly cleaned and dried before the experiment starts.
• The enzymes and chemicals used for the experiment should be of standard quality.
• Always use distilled water to make the solutions.
• Chemicals and solutions should be prepared carefully in order to avoid any wastage.

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 SECTION – B
SPOTTING

EXP: 6 STUDY OF ADAPTATIONS OF FLOWERS POLLINATED BY DIFFERENT AGENCIES

(WIND, INSECT, AND BIRD)

AIM:
To study adaptations of flowers pollinated by different agencies (wind, insect, and bird).

REQUIREMENTS:
Compound microscope, hand lens, forceps, needle, slides, cover slip, fresh flowers of maize or grass,
Salvia, Bignonia, Vallisneria, etc.

PROCEDURE:
i. Select atleast one flower for each category (wind, insect, water and bird).
ii. Observe its morphological characteristics. Note down the adaptations of the flower in its
structure to facilitate pollination by external agencies.
iii. Dissect it out to see its male and female reproductive parts.
iv. Dust the pollens on a drop of water on slide.
v. Now, observe the characteristics of pollens under microscope.
vi. And draw a labelled diagram of each category.
OBSERVATIONS:
1) Anemophily (Pollination by Wind)

A. Maize (Zea mays)

i. Maize plant bears male and female flowers separately on the same plant. Male flowers are
terminal whereas, female flowers are axillary in position.
ii. Flowers are small, inconspicuous, colourless, odourless and nectar less.
iii. Both stigma and anthers hang outside the perianth (exerted in position).
iv. Pollens are light, small and dusty. They are produced in large numbers.
v. Stigma is hairy, feathery, branched and sticky to catch wind dispersed pollens.
B. Pinus (Pine)
i. The pollens are produced in huge numbers.
ii. They are light weighted.
iii. The pollen possess wings, which take the moff along with wind to far-off places.

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 C. Grasses (Wheat and Rice)
i. Flowers are not brightly coloured or showy and are inconspicuous in nature.
ii. They do not bear scent and dot not produce nectar, but produce light weighted pollen
grains in large numbers.
iii. Stigma is feathery to catch pollen.
2) Entomophily (Pollination by Insect)

Salvia
i.The flowers are showy, brightly coloured and attract insects like honeybees.
ii.They are borne on verticellaster inflorescence to become conspicuous.
iii.Each flower has two epipetalous stamens in which stamen is attached to the petals by
filaments
iv. In older flowers, the style brings the stigma in such a position that it brushes against the
back of insect and collects pollen grains brought by the insect from a young flower.
3) Ornithophily (Pollination by Birds)

Bignonia
i. The flowers are big and brightly coloured red, orange, yellow or blue.
ii. The flower parts are thick and leathery especially corolla.
iii. The flowers produce abundant nectar and may also have certain edible parts. Birds visit
flowers for feeding on this nectar.
iv. The flowers are usually odourless (without any fragrance or scent).
RESULT:
The different types of flowers adapt themselves according to their habitat, availability and the type of
pollinators. In this way, they show great variations in their morphological features along with special
adaptive features.

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PRECAUTIONS:
• The fresh flowers should be taken to study the morphological features.
• The pollens should be gently dusted on slide with a drop of water before observing them under
the microscope.

EXP: 7 STUDY THE POLLEN GERMINATION IN STIGMA THROUGH PERMANENT SLIDE

AIM:
To study the pollen germination on stigma through permanent slide.

REQUIREMENTS:
Permanent slice of pollen germination with stigma, compound microscope-drawing sheets, pencil
eraser, etc. for temporary Side preparation:5-6 excised styles with stigma of Petunia/ grass / maize
sunflower, beaker, water, slides, cover slips cotton blue stain, brush and needle.

PROCEDURE:
Using Temporary Slide:
i. Place the freshly excised stigma and styles in boiling water in a beaker for 5-10 minutes. This
will soften the tissues and also prevent them from drying.
ii. Add cotton blue dye using a dropper onto the tissue placed on a glass slide
iii. Wait for 3-5 minutes and then wash-off the excess stain with water
iv. Mount one stigma in a drop of glycerine on a slide.
v. Place a cover slip on the stigma and gently press the cover slip on the material.
vi. Place the temporary slide under a compound microscope and observe initially under low
magnification followed by higher magnification.
vii. Draw a well-labelled diagram of the structures seen.

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Using Permanent Slide:
i. Pick up a good permanent slide of pollen germination.
ii. Place it under low power of microscope and observe the slide.
iii. Focus structures seen in low power, then in high power to have clear view of details of pollen
tube.
iv. Draw a well-labelled diagram of the structures seen.

OBSERVATIONS:
The slide shows the presence of many germinating pollen grains on the stigma of the carpel. The pollen
tubes of the germinating pollen grow through the style, transversing the tissues of the style. The slide
shows many such growing pollen tubes of varied length.

RESULT:
As the pollen tube grows into style, it reaches to the ovule and carries the two male gametes; one to
the egg cell and another to secondary nucleus for fertilisation to occur inside the ovule.
PRECAUTIONS:
• The prepared permanent slide should be handled carefully.
• Draw proper figures of the structures as seen under low and higher m gnifications of the
compound microscope.
• Label the figures accurately.

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 Exp: 8 IDENTIFICATION OF STAGES OF GAMETE DEVEL OPMENT
AIM:
To identify the stages of gamete development, i.e. T.S. of testis and T.S. of ovary through permanent
slides.

REQUIREMENTS
Permanent slides of T.S. of testis and ovary, light and compound microscope, notebook, pencil,
eraser, lens cleaning fluid and cleaning paper.
PROCEDURE
1. First, clean the slide and microscope with the help of lens cleaning paper and cleaning fluid
2. Fix the slide on the stage of microscope.
3. Observe the slide of testis and ovary first under the lower magnification and then under higher
magnification.
OBSERVATION
T.S. of Mammalian Testis

Identification Characteristics
i. The seminiferous tubules are easily seen in cross-section of testis.
ii. The presence of larger cells called Sertoli cell is the characteristic feature of this slide. These
cells provide nutrition to the developing spermatozoa.
iii. The presence of interstitial cells or Leydig's cell is evident.
General Characteristics
i. The transverse section of testis shows many semini ferous tubules embedded in interstitial
tissues. These appear as coiled, circular structures.
ii. Each seminiferous tubule is lined on its inside by germinal epithelium which divides mitotically
to produce male germ cells called spermatogonia.
iii. Various stages of development of sperm from spermatogonia are seen from periphery to the
lumen of seminiferous tubules.
iv. All these stages are clearly visible in a transverse section of testis. The mature sperms are
present in the centre and primary spermatocyte in the middle layer.
v. Sertoli cells are prominent pyramid-shaped cells found in between the germinal cells. These
cells provide nutrition to the developing spermatozoa.
vi. Leydig cells are seen in the interstitial tissue. They produce the hormone called testosterone
responsible for development of male secondary sexual characters.
T.S. of Mammalian Ovary

Identification Characteristics
i. The presence of Graafian follicle is characteristic feature of mammalian ovary.
ii. Each Graafian follicle contains one ovum and a cavity called antrum.
General Characteristics
i. The section of mammalian ovary shows the mass of Primordial Interstitial follicle cell mass
Antrum follicle tissues lined up by germinal epithelium. Ovarian stroma
ii. The stroma is made up of connective tissue, blood vessels and nerve fibres.
iii. Various circular, structures seen in stroma are graffian follicles.
iv. In the later stages of follicular which separates the ovum and the cells around it except at one
point.
v. As this cavity enlarges, Graafian follicle becomes ready to release ovum.
vi. The mature follicle ruptures and releases ovum. The ruptured follicles form corpus luteum and
release progesterone.
vii. All stages of oogenesis cannot be seen at a same time, however, they can be seen when an
ovary is observed for entire duration of menstrual cycle
RESULT:
The transverse sections of testis and ovary of mice clearly show the developmental stages of sperms
and ova by the process of gametogenesis.
PRECAUTIONS:
• The prepared permanent slide should be handled carefully.
• It should be observed initially under the low and then under the higher magnification of the
microscope.
• The diagram of the structures seen in T.S. of ovary and testis should be drawn and should be
labelled properly in order to develop understanding of the detailed structures.
 EXP: 9 STUDY OF MEIOSIS IN ONION BUD CELL or GRASSHOPPER TESTIS USING PERMANENT
SLIDE
AIM:
To study the meiosis in onion bud or grasshopper testis using permanent slides.

REQUIREMENTS
Permanent slides of all the stages of meiotic division and compound microscope.

PROCEDURE
1. Focus the slide under low power (10x) of a compound microscope.
2. Observe the dividing cells carefully.
3: Look for the nucleus, chromosome, etc. in each cells.
4. Use pointer in the objective lens to point out a specific stage of meiotic cell division to be seen in
high power.
5. Observe the stage of the cell pointed by pointer under high power (45x).
OBSERVATIONS
Different stages of meiosis have different special features. These features are summarised with proper
labelled diagram as given below:
MEIOSIS-I
A. Prophase-I
It is a complex phase characterised by number of events. It is divided into five main sub-stages:

a) Leptotene
i. Nuclear membrane and nucleolus are not clearly visible. Fine network of thin
chromatin threads is seen.

ii. These are chromatin fibres in condensed form called chromosomes.

b) Zygotene
i. Each pair has two chromosomes similar in their length and their centromere position.
ii. Chromosomes become more distinct as they become much shorter than before.

c) Pachytene
i. The homologous pairs are clearly visible.
ii. Each chromosome has two chromatids and thus each bivalent consists of 4
chromatids. Hence, the chromosomes exhibit tetrad configuration.
iii. Crossing over, i.e., exchange of chromatid segments takes place between non-sister
chromatids of homologous chromosomes

Diplotene
iv. Diplos means double, so each homologous chromosome has two chromatids that show
distinct separation from each other except at some points.
v. The attachment points of two homologous chromosomes are called chiasmata.

vi. These chiasmata represent the site of crossing over.

d) Diakinesis
i. Bivalents condense further during this stage and appear to be more shortened, thick
and prominent than before.
ii. Chiasmata are clearly visible.
iii. All homologous pairs appear in a scattered form within the cell.
iv. Nuclear membrane and nucleolus have disappeared completely.
v. Spindle formation can be seen in its early stages.

B. Metaphase-l
i. The homologous chromosomes are still in pairs and are arranged along the equatorial
plane of the cell.
ii. At this stage, number of bivalents can be counted.
iii. Chiasmata may still be seen in few bivalents.
iv. The spindle fibres attach themselves to the centrosome of each chromosome pair.
 C. Anaphase-I
i. As a result of shortening of spindle fibres, the paired chromosomes start separating.
ii. At the end of anaphase-l, the chromosomes assemble at two poles.
iii. This results into the reduction of chromosomes number to half.
iv. Each chromosome has two chromatids at this stage.

D. Telophase-l
i. Chromosomes present at the two poles appear decondensed.
ii. The nuclear membrane is formed around the two new daughter nuclei.
iii. Nucleolus also reappears.
iv. Thus, each nucleus formed has half number of chromosomes as compared to the
nucleus of the parent cell

MEIOSIS-Il
Meiosis-Il is similar to mitosis without duplication of DNA. It is divided into following stages:
A. Prophase-Il
i. The chromosomes reappear as distinct rod-shaped or thread-like chromatin fibres
ii. Each chromosome has two chromatids.
iii. Nuclear membrane and nucleolus start disappearing.
iv. The chromosomes become short by coiling and condensation.
 B. Metaphase-Il
i. This phase is similar to that of mitotic division.
ii. The chromosomes having two chromatids attached at the centromere are observed
arranged at the equatorial plane of the cell.
C. Anaphase-Il
i. The centromere of each chromosome divides into two so that each chromatid gets its
centromere.
ii. Shortening of the spindle fibres occurs so that chromatids are pulled apart towards their
respective poles.
iii. The two chromatids of each chromosome after separation appear to lie at the two poles
of the cell.
D. Telophase-Il
i. The chromatids (now chromosome) on their respective poles, now uncoil and form the
chromatin network again.
ii. Nuclear membrane and nucleolus are reformed.
iii. four haploid nuclei are seen in each cell (male or female gamete)

RESULT:
The slides under observation revealed all the characteristic features of the meiotic cell division
occurring in the onion bud or grasshopper testis.
PRECAUTIONS:
• Handle the permanent slides cautiously so that they do not break.
• Focus each slide first under 10x magnification of light microscope and then under 40 x
magnification to get the better view of dividing cells.
EXP: 10 STUDY OF TS OF BLASTULA THROUGH PERMANENT SLIDE
AIM:
Study of TS of blastula through permanent slide
REQUIREMENTS:
Permanent slide of T.S. of blastula, compound microscope, lens cleaning fluid and paper, pencil
eraser.
PROCEDURE:
i. Select a good prepared slide of T.S. of blastula and focus under 10x magnification under light
microscope.
ii. Now, study the structures and draw the different layers of cells seen.
iii. And finally compare these structures with the labelled diagrams given ahead and identify the
maccordingly.

OBSERVATIONS:
The transverse section of blastula shows the following features:
i. It is a spherical mass of cells.
ii. The outermost layer, i.e. zona pellucida followed by a layer of trophoblasts is clearly seen.
iii. Within the envelope, a fluid filled cavity called blastocoel is commonly found.
iv. Mass of cells inner to the trophoblast is called inner cell mass. Inner mass cells form the
embryo.
v. The blastocyst has embryonic or animal pole which is found opposite to the abembryonic pole.
RESULT
The transverse section of blastula clearly shows all the characteristic features of blastula stage of a
mammal.
PRECAUTIONS
• Objective and eye lenses should be cleaned before viewing the slide.
• The slide should be centrally placed so that it can be focused properly under the low and high
powers of compound microscope in order to enhance the clear visibility of all cells.
 EXP: 11 STUDY OF MENDELIAN INHERITANCE USING SEEDS OF DIFFERENT
COLOUR/SIZES OF ANY PLANT
AIM:
Study of Mendelian inheritance using seeds of different colour / sizes of any plant
REQUIREMENTS:
Pea seed sample, enamel tray, petridish, napkin, notebook, pencil/pen.

PROCEDURE:
i. Collect 100 pea seeds of different shapes and sizes and those varied in colour in an enamel tray.
ii. Put 50 round seeds in one petridish and 50 wrinkled seeds in the other to represent male and
female gametes respectively. Let the round seed be indicated by 'R' and wrinkled seed by 'r'.
iii. Take a seed from each container and place them together on the napkin.
iv. Just like the previous step, continue to pick seeds and arrange them in pairs. Thus, 50 pairs of
seeds are obtained representing the 50 heterozygous F, -progeny.
v. Note that all the F,-individuals are represented by one round and one wrinkled seed.
vi. Note the genotype RR or Rr or rr of each pair and their possible phenotype.
vii. Calculate the genotypic and phenotypic ratios of your pooled data.
OBSERVATION:

Record the result in the following table:

S.no Character of Total number of Number of seeds Approximate ratio
seed seeds observed with contrasting
studied form of traits
1) Seed shape 106 80 : 26 3:7:1
Round : wrinkled
2) Seed colour 170 83 : 27 3:7:1
Yellow : green
PRECAUTIONS:
• Large sample of seeds should be collected to minimise the error.
• The contrasting form of the trait should be observed carefully.

EXP:12 STUDY OF PREPARED PEDIGREE CHARTS OF GENETIC TRAITS SUCH AS ROLLING

OF TONGUE, BLOOD GROUPS, WIDOW'S PEAK AND COLOUR BLINDNESS.
AIM:
To Study of prepared pedigree charts of genetic traits such as rolling of tongue, blood groups, widow's
peak and colour blindness.
REQUIREMENTS:
Questionnaire about a particular disorder, a family with genetic disorder for more than one
generation,
paper, pencil, etc.
PROCEDURE:
The careful examination of chart would suggest whether the gene for the character is
i. Autosome-linked dominant or recessive.
ii. X-chromosome linked dominant or recessive.
iii. Y-chromosome linked or not.
Explanation of Each Trait
A. Autosome Linked Dominant Traits
i. The encoding gene of this trait is present on any one of the autosomes.
ii. The mutant allele is dominant and the wild type allele is recessive for such traits
iii. In this pedigree chart, the female being interviewed is exhibiting the trait and is indicated by an
arrow mark in the chart.
iv. Transmission of trait occurs from any of the parent. is
v. Male and female are equally affected. e.g., Widow’ Peak.
Widow's Peak
It is a 'V'-shaped hairline across the forehead. It is a dominant trait. Thus, both homozygous dominant
(A) and heterozygous (Aa) individuals have widow's peak, while homozygous recessive (aa)
individuals have a straight hairline.
B. Autosome Recessive Traits
i. In this, mutant allele is recessive to its wild type allele.
ii. These traits occur in equal proportions in male and female siblings, whose parents are carriers
ofthe allele, i.e., they have only a single copy of the allele and therefore they are normal.
iii. The siblings are homozygous for the defective allele, but their parents, though some may
appearnormal are obviously heterozygous, i.e., are merely carriers of the trait.
iv. The marriage between man and woman genetically related with each other, occasionally
results inthe appearance of such traits, e.g., albinism and rolling of tongue.
The Rolling of Tongue
Some persons are able to roll their tongue in U-shape. The inability to roll the tongue is caused by an
autosomal recessive allele "a'. Thus, both homozygous dominant (AA) and heterozygous (Aa)
individuals are able to roll the tongue, while homozygous recessive (aa) individuals are unable to roll
their tongue in this way.

X- Linked Recessive Traits

i. These are the traits whose encoding gene is present on the X-chromosome and its mutant
allele is recessive to its wild type allele.
ii. The females expresses the trait only when they are homozygous for the mutant allele
whereas, males does so even when they are hemizygous for it.
iii. About half of the sons of the carrier (heterozygous for the trait) females are affected.
iv. In case of homozygous females showing the trait, 50% of her daughters and all of her sons
are likely to be affected. Therefore, it can be concluded that males are most affected
population in this category.
v. Affected persons are related to one another through the maternal side of their family.
vi. Any evidence of male to male transmission of the trait rules out the X-linked inheritance.=
vii. Examples are haemophilia and red-green colour blindness.
Colour blindness

The colour blindness is a sex-linked recessive disorder of humans. In this, the affected individual is
unable to differentiate between red and green colours. It results in the absence or malfunctioning
of one or more of the three types of cone cell responsible for colour vision.
Pedigree analysis of colour blindness

BLOOD GROUP INHERITANCE

i. Blood group inheritance is independent of sex of organism.

ii. The ABO blood groups are controlled by gene I which have three alleles IO, IA, IB, I
iii. IA and IB are completely dominant over IO, IA, IB, are dominant
OBSERVATIONS:
The integrated families showed the characteristics symptoms of the genetic disorder inferred in
studies undertaken for the pedigree analysis.
RESULT:
The families studied showed the transmission of traits responsible for genetic diseases in the
successive generations in each category.

EXP 13 CONTROLLED POLLINTION – EMASCULATION, BAGGING, TAGGING

AIM:
To perform emasculation, bagging and tagging for controlled pollination.
PRINCIPLE:
The plant breeding is a science of changing and improving the heredity of plants and production of
new varieties which are far better than the original type. Artificial hybridisation is the phenomenon
used by plant breeders for the crop improvement programmes.
Steps involved in plant breeding programme are:
i. Collection of germplasm.
ii. Selection of parents with desired characteristics.
iii. 3. Cross hybridisation among selected parents which involves emasculation, tagging, bagging
and 0lastly controlled pollination.
iv. Selection and recombination of superior recombinants.
 v. Selection and testing of new superior recombinants.
vi. Testing, release and commercialisation of new cultivar.
There are three methods which are involved in the process of controlled pollination.
Tagging:
The tag is fixed to the emasculated flower mentioning the name of the parent plant, date of
emasculation, controlled pollination and the name of the plant breeder.
REQUIREMENTS:
Ornamental plants/wild plants bearing large bisexual flowers, magnifying lens, tweezers, small sharp
scissors, brush, alcohol, paper bags, rubber bands, paper clips and tags.
PROCEDURE:
The steps given below are sequentially followed to perform the experiment of controlled pollination.
a) Emasculation:

i. It is the process of removal of anther before its dehiscence, without affecting female
reproductive system, to avoid self-pollination.
i. This method is adopted for the plants having large sized flowers, e.g. cotton, pea, etc.
ii. Emasculation is done with the help of pocket lens, forceps, needle, scissors, scalpel camel hair
brush.
b) Bagging:

• The emasculated flower is immediately enclosed in a bag made of butter paper or polythene to
avoid pollination by unwanted pollen. This process is called bagging.
 c) Cross-Pollination:

• As we get the female flower we dust them with the pollens of desired planton the stigma with
sterile camel brush. This way we ensure controlled cross-pollination.
d) Tagging and Labelling

• After performing cross-pollination, tagging is done on the plant. The tag includes all necessary
information about the experiment like plant variety of male and female parent, date of
pollination, etc.
OBSERVATION:
After the emasculation, bagging and tagging is done, the plant is kept under undisturbed observation
and proper care of experimental plants is taken till seed setting and harvesting. This ensures the
success of the experiment.
RESULT:

By the process of emasculation and cross-pollination under controlled conditions, the female plant will
produce seeds of the desired characteristics
EXP 14 IDENTIFICATION OF DISEASE CAUSING ORGANISMS
AIM:
To identify common human disease causing organisms like Ascaris, Entamoeba, Plasmodium and
ringworms through permanent slides or specimens.
REQUIREMENTS:
Permanent slides of Plasmodium, Entamoeba, specimen of Ascaris and ringworm fungi(Microsporum,
Dermatophyton, Trichophyton, etc.) and a compound microscope.
PROCEDURE:
1. Study the permanent slides under compound microscope.
2. Note down the identification characteristics of organisms in the practical record book.
OBSERVATIONS:
a) Entamoeba histolytica

Characteristics
i. It is unicellular protozoan and irregular in shape due to the presence of pseudopodia.
ii. It has single big nucleus, placed eccentrically in the cell.
iii. The cell shows the presence of few food vacuoles, but contractile vacuoles are absent.
Interactions with Host
i. It is an endoparasite of human intestine and causes amoebic dysentery or amoebiasis
ii. Source of infection is through the contaminated food and water.
iii. The parasite feeds on RBCs by damaging the wall of large intestine and reaching the blood
Symptoms
i. Person affected with Entamoeba complaints of severe abdominal pain.
ii. Patient passes repeated motions filled with blood and mucus.
b) Plasmodium vivax
Interactions with Host
i. It is an endoparasite seen easily within the RBCof the infected person.
ii. The Protozoa is known to cause malaria in humans.
Characteristics:
It is a unicellular organism.
It has a big vesicle with cytoplasm accumulated at one place containing nucleus, which looks like a bag.
Symptoms
i. The infected person suffers from headache, muscular pains and high fever, 102° to 103°F
ii. The person suffers from alternate chills, shivering and high fever.
iii. Sleeplessness, restlessness and loss of appetite
c. Ascaris lumbricoides

Characteristics
i. The animal shows sexual dimorphism. The female is longer than male.
ii. Both the ends are pointed with the posterior end of male is curved ventrally.
iii. Single longitudinal lines are present on dorsal, ventral and other two lateral side throughout
the body. 0Lateral lines are more distinct than other lines.
iv. Ascaris is an endoparasite of small intestine of human beings causes Ascariasis
Symptoms
i. Ascaris worm infests a single host and obstructs the intestinal passage thereby causing
abdominal discomforts.
ii. The sufferer complaints of colic pain.
iii. There is impaired digestion, vomiting and diarrhoea in severe infestation.
iv. Person loses weight and feels tired and lethargic.
d. Microsporum
Characteristics
i. Ringworm is an infectious disease caused by fungal infection of dermatophytes.
ii. On keratinised tissue, it produces fronds of hyphae which are waxyor cotton-like.
Interactions with Host
i. The physical contact with infected persons, or their belongings may spread infections. Usually
scissors, clips remain contaminated with fungal spores and cause infection in healthy persons.
ii. Symptoms It infects the keratinised areas of the body (hair, skin, scalp and nails) and the area
gets more red on periphery than in centre creating ring-like appearance.

Symptoms on Scalp
• Yellowish cup-like crust appears on scalp. Hair become lust less and grey. In severe cases, it
leads to baldness.
Symptoms on Skin
• Round ring-shaped growing lesions appear on any part of the skin of the body.
• These lesions are very itchy which usually appear flat and keep growing if left untreated.
Symptoms on Foot
• It appears specially in between the toes because of the presence of moisture. A watery fluid
leaks out from the infected areas.
• Cracking/scaling of skin of foot occurs due to the severe infection.
Symptoms on Nails
• Nails show discolouration, become chalky and thickened.
PRECAUTIONS:
• The specimens should be examined carefully with naked eyes and with the help of hand lens.
• Prepared permanent slides should be thoroughly observed under microscope to see its
diagnostic features.
• Make your observation first under low power followed by higher magnification.

EXP: 15 MODELS SPECIMEN SHOWING SYMBIOTIC ASSCIATION IN ROOT MODULES OF

LEGUMINOUS PLANTS, CUSCUTA ON HOST, LICHENS

Aim:
To observe model specimen showing symbiotic association in root modules of leguminous plants,
cuscuta on host, lichens
Requirement-model/specimen/symbiotic association in root modules plants, cuscuta on a host and
lichen
Observation:
a) Rhizobium in root modules of leguminous plant.

i.
Rhizobium bacteria are present in root modules of legume plants and form a symbiotic
relationship, where both are benefited from each other.
ii. Rhizobium convert the atmospheric Nitrogen into soluble nitrates can be used plant for
growth development.
iii. Bacteria receives nutrients continue their growth.
b) Cuscuta with host

i. Cuscuta commonly called dodder/ a marbel and live as stem ecto parasite o other plants.
ii. Cuscuta has no fully expanded form of leaves and has no chlorophyll.
iii. The stem of cuscuta is thin and slender shaped.
iv. Stem of cuscuta fixes itself to the stem host plant with special structures called “haustoria”.
v. Haustoria absorbs the water and other solutes.
vi. Cuscuta can weaken/kill host plant.
c) Lichen

i. It represents the symbiotic relationship/association between fungus and algal.

ii. The algal component is known as phycobiont and fungal component is known as mycobionts
iii. Algae prepare food for fungi, fungi provide shelter and absorb minerals, nutrients.
iv. They grow on lands, rocks, tree trunks.
v. They are pollution indicators.
EXP: 16 Flash cards models showing examples of homologous and analogous organs.
Aim:
To study homology and analogy with the help of models/charts of animals and
models/charts/specimens of plants.
Theory:
In animals, organs that are functionally dissimilar but anatomically, or structurally, similar are called
homologous organs. Different modes of life have created the differences, i.e., modified the organs to
enable them to survive. Analogous organs
are those which are functionally similar but structurally dissimilar.
Materials Required:
Models/charts of animals and models/charts/specimens of plants.
Observations:
a) Homologous Organs:

i. The organisms which show similar anatomical structures, but differ in the morphological
structures and functions. These shows divergent evolution.
ii. By externally examine the wings of the mammal, like bat and the forelimb of aman, no
similarity is found. But after examining the bones one by one, it is found that each of them has
arm bone (humerus),hand bones (radius-ulna), wrist bones(carpals), palm bones (metacarpals),
and fingers (phalanges).
iii. In terms of proportions of growth of each constituent bone, there are differences. For example,
the fingers of bat are much longer while the comparative study suggests is that basically the
forelimbs of these two creatures are made up of the same parts, that is, they are anatomically
similar.
iv. These organs need not perform the same function, as we see that bat uses it for flying and man
uses it for handling tools. Hence, the forelimb of man and the wing of bat are homologous
organs. Similarly, forearms of cat and man are homologous.
Ex. Flipper- Whale, Wing- bat, Fore limb- horse, Paw- cat, Hand-human
Thorn- bougainvillaea, Tendril- Cucurbita
b) Analogous Organ:

i. The organisms which show similar morphological structures and functions, but differ in the
anatomical structures. These shows convergent evolution.
ii. The internal structure of the wings of butterfly, its preserved specimen, the shape and size
are observed. It is found that it is membranous and is made up of thin cuticle.
iii. There are veins in the wing but there is no skeleton.
iv. The wings of the preserved specimen of a bat and a bird are examined. Skeletal support is
found. It shows that the basic structures of wings of butterfly, bird and bat are different. In
other words, they are anatomically different, although externally they lookalike. Wings in
these animals are used for Flying .
v. Such organs that differ an atomically and in embryonic mode of origin but perform similar
function are said to be analogous organs.
Ex: Wings of butterfly and wings of a bird.
Flippers of Penguin and Dolphin
Eye of an Octopus and a mammal (retinal position)
Sweet potato (root modification) and potato (stem modification)