Study of the Microscope

Aim - To study the parts of a microscope

SIMPLE DISSECTING MICROSCOPE

A simple dissecting microscope is needed for small objects for dissection or detailed study. It is a
single lens system. It consists of the following parts:
(a) A base or foot made of heavy material that supports the weight of the microscope
(b) A vertical stand with folded arms on which the lens is mounted
(c) A focusing limb that can be brought up and down with the help of an adjustment screw
(d) A glass stage with clips where the object is fixed at a required position
(e) A simple convex lens system for magnifying the object
(f) A reflecting mirror fitted below the stage for adjustment of light.

Procedure to be followed -
The object to be examined in the dissecting microscope is placed on the glass plate under the
lens. Then the object is illuminated suitably by adjusting the mirror. The lens is moved to align over
the object. One eye is placed over the upper side of the lens and the adjustment screw is moved
up and down until a sharp image of the object is obtained.

COMPOUND MICROSCOPE: Used for higher magnification

Principle
Consists of two sets of lenses
First lens - objective (close to object)
Second lens - eyepiece
The magnification which objective produces at a fixed distance is called magnifying power
The total image enlargement or the total magnification produced with the microscope is the
product of the primary and secondary (objective and eye price) magnification.

Parts of a compound microscope:

1)Foot - The microscope is built around a strong basal foot and a vertical limb. The foot supports
the limb and rests on the table.
2)Body Tube - The objectives are screwed to the revolving nose piece which is attached to the
lower end of the body tube. At the upper end of the tube is the eyepiece. The body tube itself
though incapable of lateral movement can be moved along its axis to bring the lens into
adjustment using two knobs- one coarse and another fine. The tube and its attachment are
connected to the base of the instrument using a hinged joint. This helps in the convenient handling
of the instrument.
3) Stage - Below the objective in the centre of the microscope, is the stage on which the
microscope slide along with the object can be kept. The stage is attached to the limb using a
broad rigid bracket. The stage is provided with two clips to hold onto the slide.

4) Substage - A movable or fixed sub-stage is placed directly below the stage. It is provided with
an iris diaphragm and condensed lens. Iris diaphragm is a wheel-shaped metal disc that regulates
the aperture. It controls the intensity of light entering the condenser lens. The condenser, a system
of two or more lenses, is used to increase the intensity of illumination of the object. It gathers all
the possible light rays, condenses and sends them to the object. The condenser can be raised or
lowered using a rack and pinion arrangement.

5) Mirror - At the base, below the condenser, a Plano-concave mirror is fixed which gather light
rays and sends them to the condenser.

PRECAUTIONS
When not in use, keep the microscope in its cabinet
Always keep the microscope in an upright position while moving it from one place to another
Never touch the lens with your hand. Before and after the use, all lenses and the body of the
microscope should be cleaned. The lenses should be cleaned with tissue paper or a mulling cloth.
Always bring the object into focus-first in low power and then in high power, do not use higher
power than is necessary
Never turn the coarse adjustment knot while using the high-power objective
Observe the object with both eyes open; get accustomed to using either eye
Never focus downward while looking through the eyepiece
Take great care to focus accurately to avoid eye strain
- See that the microscope is clean (free from traces of water, fixatives, stains and reagents) and
dry and in good order when you put it rack.
 Test for Starch - Food Tests
Experiment 1 - Starch
Aim - To test the distribution of starch grains in a potato tuber
Requirements - Potato tubes, razor, slide iodine solution, microscope, Petri plate
Procedure - Make thin sections of the pulp of a potato tuber and mount them in an iodine solution
Observation - Starch grains take blue stain
Result - Starch grains in potato tuber can easily be located by blue-black colour

Experiment 2 - Reducing sugars

Aim - To test the presence of reducing sugar (e.g., glucose) in the given solution.
Requirements - Test tubes, Fehling reagent or Benedict reagent, spirit lamp
Procedure - Take approximately 3 ml of a given solution in a test tube, and add an equal quantity
of Benedict reagent or Fehling reagent to it. The test tube is shaken well and then put in a water
bath till the boiling of the solution.
Observation - A brick-red brown or green precipitate appears
Results- The precipitate indicates the presence of reducing sugar

Experiment 3 – non-reducing sugar

Aim- To test the presence of non-reducing (e.g., sucrose) in a given solution
Requirements. Test tubes, hydrochloric acid, Sodium bicarbonate, Benedict reagent, sucrose
solution spirit lamp
Procedure - Boil a little amount of given solution (sucrose solution) with dilute hydrochloric
neutralise with sodium bicarbonate, then test with Benedict's reagent

Experiment 4 - Lipids (fats)

Aim - To test the presence of lipids (fats)
Requirements – Paper strip test
Procedure – Coat a strip of paper in a given solution
Observation- The paper turns translucent

Result - Appearance of a brick-red colour on the surface of the substance indicates the presence
of protein (Proteins show this reaction due to the presence of tyrosine)
Caution - Millor's reagent is poisonous. Do not boil it for long, inhale its fumes or get it on your
skin.
Experiment 6 - Protein
Aim - To test the presence of proteins by using Biuret's reagent
Requirements - Test tube, protein suspension, Biuret reagent
Procedure - Take about 3ml of a solution of the suspension of protein in a test tube and add a few
drops of Biuret reagent down the side of the test tube. Do not heat.
Observation - A blue ring appears at the surface of the solution that on shaking disappears and
the solution turns purple
Result - Presence of protein is confirmed (Protein show this reaction due to the presence of
peptide bonds in their structure)
Permeability of Cell Membrane
Aim: To demonstrate the effect of heat on the permeability of the cell membrane of beetroot cells
Requirements: Test tubes, beaker, measuring cylinder, spirit lamp, mat, tripod and gauze, cork
borer, razor blade, thermometer, watch, forceps, marker
Procedure: Select a large fresh beetroot and wash it in running water. Cut a few cylinders from it
with a cork borer. Make all cylinders of the same length with the help of a sharp blade. Wash
cylinders in running tap water to remove injured cells. Using a spirit lamp, a tripod and a gauze,
heat 200 ml of distilled water to 100℃ in a 250 ml beaker. Now take four test tubes and mark them
A, B, C, D. Put 15 ml of boiling water in test tubes A and C and freezing water in D. Add distilled
water to C to drop the temperature to 50°C. Fill test tube B with distilled water at room
temperature. The water in each test tube should be at the following temp
A 100°C.
B room temperature
C 50°C
D freezing

Now put two cylinders in each test tube. After 30 minutes, shake the test tube and then remove
the cylinders. Compare the colour of A, B, C, D. (B as control)
Observation
Test Tube Temp (C) Degree of Pigment
A 100 C Max
B 28 C (rt) None
C 50 C None
D 0C None

Inference - High temperatures kill the cell membrane thereby making it fully permeable; hence at
high temperatures, maximum leaching of anthocyanin is seen.
Precautions
The beetroot should be fresh
All cylinders should be equal in dimension
Edges of cylinders must be level
Cylinders should be thoroughly washed.
The number of cylinders and amount of water should be constant
Note - The experiment can also be carried out by heating one beetroot with alcohol. It kills the cell
membrane thereby making it fully permeable and allowing max leaching
 Demonstration of Plasmolysis
Aim - To demonstrate the phenomenon of plasmolysis in plant cell
Requirements - Rhoeo discolour leaf/ onion bulb scale/ rose petal, safety blade, watch glass, petri
dish, slides, forceps, brush, needle, coverslip, salt or sugar solution of different concentrations,
microscope.
Procedure - Peel off a thin scrap from the lower surface of a Rhoeo discolour leaf. This is done by
tearing the leaf obliquely with a single jerk or scraping it with a safety blade. Mount one peel in a
drop of water on a slide and examine the cells of the peel under the microscope.
Take another peel and mount the pieces. in a drop of salt or sugar solutions of different
concentrations
Observation - Observe the prepared slides under a microscope and draw the boundaries of the
protoplasm in each case. Look for plasmolysis. What happens when a plasmolysed cell is again
put in the water?
Result - In the peel mounted in water, cells are turgid and protoplasm is closely pressed against
the cell wall. When a slightly concentrated solution is used for mounting, the cell contents withdraw
a little from the cell wall. Colourless space between the cell wall and the coloured all sap is
distinctly seen. When more concentrated solutions are used, the cell contents move appreciably
away from the cell wall and ultimately the cell contents withdraw from the cell wall and shrink into a
small, round, ball-like form. When plasmolysis cells are put in water the protoplasm of the cells
regains its original position.
Conclusion - When a peel is kept in concentrated salt/sugar solution (hypertonic solution), the cells
lose water due to exosmosis. As a result, the protoplasm shrinks leaving the cell wall in its original
position. The small space between the cell wall and the contents indicates the beginning of
plasmolysis. It is known as incipient plasmolysis. With the increase in the concentration of solution
outside, the space between the cell wall and contents increases. Finally, due to continued
exosmosis, the cell becomes fully plasmolysed. In a plasmolysed cell, the space between the cell
wall and the cell wall and the contents is filled with the hypertonic solution which diffuses through
the permeable wall of the cell.
Precautions
Always use fresh plant material
Never allow material to dry up
Observe the same cells for plasmolysis and deplasmolysis
 Cell Structure
Aim - To study the structure of a plant cell by using onion peel
Requirements – Onion, scalpel, water, safranin/iodine, slide, cover-slip, glycerine, compound
microscope.
Procedure - Take an onion bulb and strip off the transparent peel from the inner fleshy scale. Stain
the peel with the help of safranin/iodine and mount in glycine. Examine the peel under a
compound microscope.
Observation
Looking at the epidermal peel under a compound microscope the following structures can be
seen-
1) Cell wall
It is found as a protective layer all around the cell and provides shape and rigidity to the
plant cell. The cell wall is made up of cellulose. Inner to the cell wall, a cell membrane is
found
2) The cell membrane is inner to the cell wall
3) Cytoplasm
It is the jelly-like fluid protoplasmic matrix that surrounds the nucleus and constitutes the
true internal milieu of the cell. It contains several organelles such as mitochondria, plastids
etc.
4) Nucleus
The nucleus is generally spherical and controls all the processes of a cell.
 Stages of Mitosis in Onion Root Tips
Aim - To investigate the phases in mitosis using onion root tips
Requirements - Onion bulb, 125 ml bottles, small corked tube, forceps, petri dish, hot plate,
compound microscope, slides, cover clips, razor blade, watch glass, acetocarmine, hydrochloric
acid
Procedure –
1) Growing of roots from onion bulbs
Place an onion bulb on a bottle filled with water so that its base remains submerged in
water. Leave it as such for 4-5 days without any disturbance. Alternatively, onion bulbs may
also be germinated in earthen pots filled with sawdust or sand.
2) Fixation of root tips
When roots have grown 1-2 cm remove the bulb and cut off the terminal 5mm of these
roots. For fixation transfer the root tips to a small corked tube containing acetic -alcohol and
leave overnight at room temperature.
3) Preparation of smear
 a. Remove root tips with forceps by holding the cut end of the root, transfer to a petri
dish containing distilled water and wash for a few minutes to remove the fixative
b. Transfer the root tips in a watch glass containing acetocarmine stain and molar
hydrochloric acid in the proportion of ten parts stain to one part acid
c. Warm but do not boil for 6-10 minutes on a hot plate (in this process middle lamellae
break down which holds cells together)
d. Place the stained root tip on a clean slide. Cut in half transversely and discard the
farthest half of the apex
e. Add 2 to 3 drops of stain to the root tip on а slide and without interfering too much
with the arrangement of cells, break up root tip with a needle and spread out as
thinly as possible.
f. Put on a coverslip, and squash gently. If necessary, irrigate with more stain
g. Warm the sides on a hot plate for 10 seconds to intensify the staining. The slide
should be warm but not too hot to touch
h. Examine the slide under the low and high microscope and identify cells showing
different stages of mitosis.
i. Draw and label the cells showing phases of cell division
Observation - Cells in the following stages of mitosis are seen in a smear of the onion root tip
1) Interphase
a. It is a non-dividing phase of the cell cycle between two successive cell divisions
b. Chromatin fibres appear in the form of a network within the nucleus
c. Nuclear envelope and nucleolus are distinct
2) Prophase
a. Chromatin materials shorten and condense into thread-like structures called
chromosomes
b. Each chromosome consists of two chromatids, joined at a point called the
centromere
c. The nuclear membrane and nucleolus start disintegration and disappear at the end
of the prophase.
3) Metaphase
a. A bipolar spindle develops in the cell. Chromosomes become thick and the two
chromatids of each chromosome become clear.
b. Chromosomes become arranged at the equator of the spindle
c. Each chromosome gets attached to the spindle fibre by the centromere

4) Anaphase
a. The two sister chromatics of each chromosome separate from the centromere and
move towards the opposite poles
b. The daughter chromosomes (separated chromatid) appear V, J, L, I shaped
depending upon the position of the centromere
5) Telophase
a. The spindle disappears and the daughter chromosomes uncoil to form chromatin
fibres at the two poles
b. Nuclear membrane and nucleolus reappear and two daughter nuclei appear at
opposite poles
c. Cytokinesis occurs by cell plate formation between the two daughter nuclei.
Precautions
The base of the onion bulb should be in contact with water while growing the roots
Root tips should be fixed in the morning between 8 to 10 am
The slide should be warmed gently much above the flame of the spirit lamp
 Effect of CO2 on Photosynthesis
Aim - To demonstrate the effect of different carbon dioxide concentrations on the rate of
photosynthesis

Requirements - An aquatic plant (Hydrilla), beaker, funnel, test tube, distilled water, sodium
bicarbonate, physical balance, pipette stopwatch, measuring cylinders
Procedure -
1) Preparation of sodium bicarbonate solution. Dissolve 10g of sodium bicarbonate in 100 ml
of distilled water. This is 10%. Solution of sodium bicarbonate. Every 1 ml contains 0.1g of
NaHCO3.
2) Setting up of the experiment
Fill the select beaker with distilled water. Select a healthy twig of Hydrilla. Cut its stem
obliquely with a sharp blade in water. Insert the twig in the funnel in such a way that the cut
end is firmly inside the stem of the funnel but the twig is spreading in its wide mouth. The
funnel is then kept in the beaker fully immersed in water. Thereafter, a test tube filled with
water is inverted over the stem of the funnel in such a way that no water comes out of the
tube. Leave this set up in sunlight and observe the gas bubbles coming out of the cut end of
the stem. Count the number of gas bubbles that evolved per minute. Then add 10 ml of
10% solution of NaHCO3 in the experimental set-up and again count the number of gas
bubbles evolved per minute.
Observation
No. Qty of NaHCO3 in ml No. of bubbles evolved
1 0 16
2 10 20
3 20 25
The number of air bubble production increases by the addition of sodium bicarbonate solution

Result - The rate of photosynthesis increases with the addition of sodium bicarbonate that
increases the supply of CO₂
Precautions-
1) Ensure that water does not flow out of the test tube remain fully and should remain filled
with water (the rate increase is up to a certain limit).
 T.S. of Monocot and Dicot Root
TS of Dicot root
Cut a thin transverse section of the material
Stain with safranin - fast green combination, mount in glycerine and study.

Staining and Mounting

Select the thinnest sections and stain them in the appropriate combination. For general anatomical
work safranin - fast green (or light green aniline blue) combination is most suitable.

(4-5 min) Stain with safranin

Wash repeatedly with water

Destain with acid water (if needed)

Wash thoroughly in water to remove traces of and

Stain with fast green (a light green or aniline blue) for about a minute

Wash thoroughly in water

Mount in glycerine.

A transverse section of a dicot root shows the following structures-

1. Epiblema (Piliferous layer) - This is the outermost layer of thin-walled cells;

the outer walls of the cell proliferate outwards to form unicellular root hairs.

2. Corten - It is undifferentiated and consists of several layers of closely

arranged thin-walled parenchymatous cells
 3. Endodermis - It is made up of a single layer of closely packed barrel-shaped
cells. This layer separates the vascular tissue from the cortex. The radial
and tangential walls of endodermal cells have Casparian thickening.

4. Pericycle - It is a single layer of thin-walled cells inside the endodermis

5. Vascular Tissue - It shows, radial arrangement with usually 4(2-6) strands of

xylem alternating with phloem stands. The xylem is an exarch
A parenchymatous conjunctive tissue is present between the phloem and
xylem strands.

6. Pith - It occupies a small area in the centre of the root. It is composed of

thin-walled parenchymatous cells

Points of Identification
 Unicellular hairs present, on the epiblema
 Vascular bundles are radial and exarch
 Xylem stands are two to six
 Vessels are angular or polygonal in transection
 Pith is absent or poorly developed.

TS of monocot root
Cut a thin transverse section of the material stain, with Safranin fast green combination mount in
glycerine and study.

Transection of a young the following structures maize root shows the following structures-
1. Epiblema – This is the single outermost layer of thin-walled cells that give rise to unicellular
root hairs.
2. Cortex - It is composed of several layers of loosely arranged, parenchymatous cells with
distinct intercellular spaces
3. Endodermis - This is a layer of compactly arranged, barrel-shaped cells with characteristic
Casparian thickenings on their radial and tangential walls. Some cells of the endodermis
placed against the protoxylem elements are thin-walled passage cells.
4. Pericycle - It follows the endodermis and is of composed a single layer of compacity
arranged thin-walled cells.
5. Vascular tissues - The vascular tissue shows the radial arrangement. There are 6-12
strands of xylem alternating with as many phloem stands. The xylem is exarch. In between
xylem and phloem strands, parenchymatous conjunctive tissue is present.
6. Pith - The pith is well developed, consisting of loosely arranged parenchymatous cells.
These cells are rich in starch grains.
Points of Identification-
1. Unicellular hairs present on the epiblema
2. Vascular bundles radial and exarch
3. Xylem strands six or more
4. Vessels appear oval or rounded in transection
5. Pith is large and well developed.
TS of Dicot Stem
Cut a thin transverse section of the material strain with safranin-fast green combination, mount in
glycerine and study.

1) Epidermis - It is the outermost layer of tangentially elongated cells - Multicellular hairs and
Stomata are present in the epidermis
2) Cortex - Composed of several layers of cells, it is differentiated into three regions-
a. Hypodermis - This is the outermost region of the context and consists of 3-5 layers of
collenchymatous cells.
b. General cortex - This is the middle zone of the cortex, composed of several layers of
parenchymatous cells with intracellular spaces.
c. Endodermis - This is the innermost layer of the cortex which delimits it from the
vascular cylinder. The endodermal cells are barrel-shaped and they accumulate
starch in a zone called a starch sheath.
3) Pericycle - The endodermis is followed by the pericycle which is in the form of patches of
sclerenchymatous cells.
4) Vascular bundles –
a. Arranged in a rung
b. Conjoint
c. Collateral
d. Open
Each bundle has outer phloem and inner xylem and a strip of cambium in between them.
These cells have thin walls and appear almost rectangular in TS.
5) Pith - Well developed and occupies the central portion of the stem. Made up of large thin-
walled parenchymatous cell's spaces with distinct intercellular spaces

Points of Identification
1) Cortex well differentiated
2) Vascular bundles conjoint, endarch, open and arranged in a ring
3) Pith well developed

TS of Monocot Stem.
Cut a thin transverse section of the material, stain with Safranin -fast green combination mount in
glycerine and study.
1) Epidermis - It is a single layer of parenchymatous cells, covered on the outer side by a thick
layer of cuticle. It contains stomata.
 2) Hypodermis - The epidermis is followed by 2-4 layers of sclerenchymatous cells which form
the hypodermis
3) Ground tissue - It extends from just below hypodermis to the centre of the stem and is
composed of parenchymatous cells.
4) Vascular bundles – Numerous vascular bundles are scattered in the ground tissue. They
are conjoint, collateral and closed. The peripheral vascular bundles are smaller and as they
pass towards the centre, they become larger. Each vascular bundle is surrounded by a
sclerenchymatous sheath. The xylem of the bundle consists of vessels, tracheids and
xylem parenchyma. There are four distinct vessels arranged in the form of '4'; the two
smaller vessels lying laterally constitute the metaxylem. Small pitted tracheids are present
in between the arm of ‘Y’. In an older vascular bundle, a schizolysigenous cavity is formed
by the disintegration of protoxylem elements. The phloem consists of a sieve tube and
companion cells. Phloem parenchyma is absent.

Points of Identification
1) Cortex undifferentiated, presence of ground tissue
2) Vascular bundles are conjoint, collateral, and are scattered on the ground tissue
3) The vascular bundle is closed and is surrounded by a bundle sheath
4) Xylem is Y shaped and endarch
Monocot root
Dicot root
 Spotting
SPOTTING SAMPLES

Agaricus (Mushroom)

I. Classification
Kingdom - Mycota (fungi),
Division – Basidiomycota
Class - Hymenomycetes
Order - Agaricales
Family - Agaricaceae

II. Diagnostic features-

1. It is an umbrella-shaped structure with a long massive stripe and a broad pileus.
2. Stripe is a thick fleshy and cylindrical structure.
3. Umbrella like pileus is present at the distal end to the strip
4. Numerous present vertically hanging gills are present on the undersurface of the pileus

Riccia (Liverwort)

I. Classification
Kingdom - Plantae
Division – Bryophyta
Class - Hepaticopsida
Order – Marchantiales
Family – Ricciaceae

II. Diagnostic features-

1. Plant body thalloid, prostate, flat, dorsiventral and dichotomously branched
2. Thallus is linear to wedge-shaped with an apical notch at the apex and thick midrib. On the
dorsal surface, the midrib is transversed by a mid-dorsal - groove.
3. On the ventral surface of the thallus rhizoids are present in the midrib region and scale at the
margins
4. Scales are multicellular
Funaria (Moss)

I. Classification
Kingdom – Plantae
Division – Bryophyta
Class – Bryopsida
Order – Funariales
Family – Funariaceae

II. Diagnostic features

1. The plant body is foliose with an erect leafy axis with a tuft of rhizoids at the base.
2. Rhizoids are slender, branched, multicellular with oblique septa
3. Axis is upright, slender and branched
4. Leaves are simple sessile with district midrib, especially arrange sporophyte

Sporophyte
1. The sporophyte is present at the tip or the archegonial branch
2. Differentiated into the foot, seta and capsule

Dryopteris (Fern)

I. Classification
Kingdom – Plantae
Division – Tracheophyta
Sub-division – Pteropseda
Class – Leptosporangiate
Order – Filicales
Family – Polypodiaceae

II. Diagnostic features

1. The sporophytic plait body is differentiated into roots, rhizome and leaves.
2. Adventitious roots are present on the underside of the rhizome
3. Leaves are large and bipinnately compound; the lower portion of the leaf is covered by their
hairy ramenta
4. Young leaves. are circinately coiled
Pinus (Pine)
I. Classification
Kingdom – Plantae
Division – Gymnosperms
Class – Coniferopsida
Order – Coniferales
Family – Pinaceae

II. Diagnostic features

1. The plant body is differentiated into root, stem and leaves.
2. Presence of long shoots and dwarf shoots
3. Leaves are needle-like
4. Both male and female cones are present
5. Male clusters replace dwarf shoots
6. Female cones occur singles and replace the long shoots

Monocot plant(bamboo)
1. The plant is attached by a tuft of fibrous adventitious roots and persists using sympodial
rhizome formed by the lower internodes of the stem
2. Stems are hollow and jointed with long internodes
3. Leaves are alternate
4. Venation is parallel

Dicot plant(petunia)
1. The stem is herbaceous, aerial, erect, cylindrical, branched, hairy, and green
2. Leaves are simple and have reticulate venation
3. Flowers are borne in a few-flowered cyme in shades of white and purple

Espongia (Common bath sponge)

I. Classification
Kingdom – Animalia
Phylum – Porifera
Class – Demospongiae
Order – Keratosa

II. Diagnostic Features

 1. The body is massive with variable shapes dark brown or black in colours
2. Several small projections made up of sponge fibres and known as convuli over the surface
3. The canal system leuconid type
4. Sexes are separate

Hydra

I. Classification
Kingdom – Animalia
Phylum – Cnidaria
Class – Hydrozoa
Order – Anthoathecata

II. Diagnostic features

1. The body is elongated and tubular, open at the distal and closed at the proximal end
2. The free distal end has a small conical hypostome with a star-shaped mouth
3. 6-10 tentacles present
4. Single large internal cavity called coelenteron present

Hirudinaria granulosa (Indian cattle leech)

I. Classification
Kingdom – Animalia
Phylum - Annelida
Class – Hirudinea
Order - Gnathobdellida
Genus - Hirudinaria

II. Diagnostic features

1. Elongated uniform body, divisible into 33 metamerically arranged segments; each segment is
divided externally by transverse grooves
2. Body distinguished into the cephalic, pre-clitellar, clitellar, middle, caudal and posterior sucker
3. Digestive fact is a straight tube
4. Excretory system consists of 17 pairs of segmented nephridia
Silkworm

I. Classification
Kingdom – Animalia
Phylum – Arthropod
Sub-phylum – Mandibulata
Class – Insecta
Order – Lepidoptera

II. Diagnostic features

1. Body differentiated into head thorax and abdomen
2. The head consists of pair of pectinate antennae a pair of compounds sucking mouthparts
3. Thorax bears 3 pairs of legs and 2 pairs of membranous wings
4. Hindwings are comparatively smaller than the forewings

Labeo rohita (rohu)

I. Classification
Kingdom – Animalia
Phylum – Chordata
Subphylum – Vertebrata
Group – Gnathostomata
Superclass – Pisces
Class – Osteichthyes
Order – Cypriniformes

II. Diagnostic features

1. Body spindle-shaped, divisible into a head, trunk and tail
2. The mouth is subterminal and remains bounded by thick and fleshy lips
3. Snout bears dorsally a pair of small nostrils
4. Lateral eyes on the head without
5. Presence of bony gill cover (operculum)
 Permanent slide spotting
1. Blood smear of human
a. The blood corpuscles are of 3 types, viz. erythrocytes (red blood cells), leukocytes
(white) blood cells) and blood platelets
b. Erythrocytes are enucleated, circular structures appear biconcave side view and are
arranged in stacks. They are devoid of cell organelles.
c. Leukocytes are nucleated cells but they lack haemoglobin. They are of two types,
granulocytes and agranulocytes
d. Granulocytes possess granules in their cytoplasm based on staining properties of
the granules, they are of three types
i. neutrophils nucleus is visible in shape and has various lobes
ii. eosinophil nucleus is composed of two or three lobes which are joined
together delicate threads.
iii. Basophil’s nucleus is s-shaped
e. The agranulocytes are devoid of granules and are of two types,
i. lymphocytes
ii. monocytes

2. Amoeba

I. Classification
Kingdom – Protista
Phylum – Protozoa
Class – Rhizopoda
Order – Lobosa

II. Diagnostic Features

a. Shapeless jelly-like mass of protoplasm that remains surrounded by a delicate cell
surface membrane and changes its shape constantly with the help of pseudopodia
b. The cytoplasm is differentiated into an outer layer of clear plasma gel called
ectoplasm and an inner mass of granular plasma known as endoplasm
c. Endoplasm contains a variable number of contractile vacuoles and a variety of food
vacuoles
d. The nucleus is embedded in the cytoplasm but occupies no fixed position

3. Saccharomyces
I. Classification
Kingdom – Mycota
Division – Ascomycotina
Class – Endomycetales
 Family – Saccharomyxetaceae

II. Diagnostic features

a. It is a unicellular saprophytic fungus of considerable economic importance
b. Cells may be globose, oval, or elliptical
c. Presence of distinct thin and firm cell wall
d. Presence of centrally located large vacuole
e. Presence of a small nucleus attached to the vacuole
f. Vegetative reproduction takes place by budding

4. Spirogyra
I. Classification
Kingdom – Plantae
Group – Algae
Division – Chlorophyta
Order – Conjugales
Family – Zygnemaceae

II. Diagnostic Features

a. Plant body filamentous and unbranched
b. Filaments uniseriate consisting of a row of cylindrical cells
c. Presence of large vacuole across which runs strands of cytoplasm
d. Cells are uninucleated the nucleus remains suspended in a central position by
protoplasmic Strands.
e. The most important feature of the cell presence of one or more chloroplasts that are
spiral and ribbon-shaped. Many pyrenoids are embedded in the chloroplasts at
regular intervals.
 Chromatography
Object – To separate plant pigments by paper chromatography.
Requirements – Spinach leaf/leaf extract, chromatographic paper, broad mouth test tube/jar,
petroleum ether, acetone, beaker, funnel, mortar and pestle, filter paper, capillary tube, rubber,
cork, blade
Procedure
1. Preparation of leaf extract. Take 10-15 spinach leaves, place them in a clean mortar, add a
little amount of acetone and reduce the leaves to a pulp with the help of a pestle. Stir well
and filter in the leaf extract.
2. Preparation of chromatographic paper step. Cut chromatographic paper into a strip to a size
that would fit easily in a test tube/jar
3. Put a circular spot of leaf extract on the paper with the capillary tube, 2 cm above the
bottom of the paper strip.
4. Allow the pigment spot to dry. Repeat the same process 5-6 times. Let the spot dry
5. Now, add about 5ml petroleum ether acetone solvent (9 parts petroleum ether: I part
acetone) in the test tube /jar, then with the help of the split cork place the spotted paper
vertically so that the spot is just above the solvent level.
6. Keep the test tube/jar airtight with the help of the cork and keep the tube undisturbed
7. The solvent shall move up the paper
8. Remove the chromatogram when the solvent reaches the top
9. Take out the chromatogram from the test tube/jar and allow it to dry
10. Since the colours of the pigments fade, examine the paper when fresh

Observation
Pigments Colour of band
Carotene Orange-yellow
Xanthophyll Yellow
Chlorophyll a Blue-green
Chlorophyll b Yellow-Green

Precaution
1. The chromatographic paper strip should not touch the walls of the test tube/jar
2. Cork should be airtight
3. The pigment spots should be placed at least 2-3 cm above the bottom of the paper strip
4. Place the spotted paper vertically, so that spot is just above solvent level
5. Do not dry strip in the sun
 Observation and Comments on the Experimental Setup
Objective
Demonstration of osmosis in the laboratory by using parchment paper.
Requirements
Sugar Solution, water, thistle funnel, parchment paper, blade, thread, beaker, stand.
Procedure
The phenomenon of osmosis can be easily demonstrated in the laboratory by using an animal
bladder or parchment paper. The membrane is tied over the thistle funnel with a long stem. The
funnel is filled with a concentrated sugar solution with the stem of the funnel partially empty and
then inverted and dipped in distilled water in a beaker.
Observation
The level of the solution within the stem of the funnel goes up steadily.
Inference
The level rise is due to the entry of water molecules into the sugar solution in the funnel through
the semipermeable membrane by osmosis. After some time, the level of sugar solution in the
stems of the funnel becomes static as there is no more entry of water molecules inside. This is
because the solution is prevented to gain more solvent molecules.

Objective - Transpiration pull

The water absorbed by root hairs passes through the cortex, passage cells and pericycle to enter
the tracheary elements of the xylem. It takes place either actively through the DPD gradient or
passively through transpiration pull. Water is lifted to leaves as a continuous unbroken column in
the tracheary elements of the xylem by one or more forces like root pressure, cohesion and
adhesion of water and transpiration pull acting simultaneously.
Experimental setup
Fill the let up the whole apparatus with water by keeping a thumb on the lower end of the capillary
tube is put in the coloured – water-filled beaker and tie the apparatus with a stand with the help of
clamps. Now insert the twig through the hole into the cork in a way that the cut end of the plant is
dipped in the water. Make the cork and hole airtight by applying Vaseline. Note the initial level of
coloured water in the capillary. Allow the experiment to stand in an open and sunny place. Note
the level at the end of the experiment.
Observation – Level of colour uses after sometime
Inference
The plant transpires and absorbs water to make up the deficit thus created. This results in pull or
suction called transpiration pull.
The transpiration pull exerts a tension on the water present in the xylem elements. This tension is
passed down to the roots. Thus, the whole column of water is lifted. The colour column is similarly
pulled upward exhibiting transpiration pull.
Ganong's Potometer

Procedure
A twig of some suitable plant cut with a sharp knife is fixed in an apparatus. The entire
apparatuses are filled with water so no air spaces are present. An air bubble is introduced into the
horizontal graduated capillary tube which is dipping into the beaker containing water. Potometers
measure the rate of water uptake.

Observation
The air bubble moves up the graduated cylinder indicating uptake.
Precautions
1. The potometer should be made completely watertight
2. The twig should be cut obliquely and underwater to avoid suction of air bubble into the twig
which will stop the absorption of water into the xylem.
 Demonstration of osmosis in plant cell
Objective
To demonstrate the osmosis by using potato osmoscope
Requirements
Potato tuber, sugar solution, water, peter dish, pins.
Procedure
Peel the outer skin of the potato tuber. Cut one end flat. From the other end scoop a cavity neatly
down to the bottom. Half fill the cavity with sugar solution and mark the level of the solution in the
cavity by piercing with a pin. Place the tuber on its flat end in a dish of pure water so that the level
of water in the dish is not higher than the sugar solution in the tuber’s cavity.
Observation
After some time, it is observed that the level of sugars solution being higher, the water moves
through and level in the tuber rises.
Conclusion
The flow of solvent is along the concentration gradient i.e. higher to lower
 Study of Different Modifications in Leaves
The leaf is a flattered, lateral outgrowth of the stem or the branch, developing from a node and
having a bud in its axil. Leaves always develop in acropetal order on the stem and are exogenous
in origin.

Parts of Leaf
A typical foliage leaf may be differentiated into 3 parts: leaf base, petiole and lamina
1. Leaf base - It is the point of attachment of the leaf to the stem, Leaf base is simple with a
pair of outgrowths called stipules
2. Petiole - It is the stalk of the leaf, which helps in the attachment of the leaf to the stem. A
leaf with petiole is called petiolate
3. Lamina - Flat expanded portion of the leaf is called lamina or blade.

Venation
The arrangement or disposition of veins and veinlets in the leaf lamina is called venation.
1. Reticulate Venation - When veins are irregularly distributed to form a network, it is known
as reticulate venation
2. Parallel Venation - When veins run parallel to each other and do not form a reticulum it is
known as parallel venation.
Simple and Compound Leaves

Simple Leaves
In simple leaves, there is a single lamina which is usually entire or sometimes incised, but the
incisions never reach up to the mid-rib
Compound Leaves
In compound leaves incisions of the lamina reach up to the midrib (rachis) and the lamina is
divided into several small segments called leaflets or pinnate. they are all easily distinguishable
from simple leaves by the absence of buds in their axils

1. Pinnately compound leaf - In this type, incisions of the lamina reach up and are directed
towards the midrib, known as rachis. Leaflets are arranged on both sides of the rachis
alternately or in the opposite manner
a. Bipinnate - A twice pinnate leaf eg Acacia
b. Tripinnate - A thrice pinnate leaf. eg Moringa
 2. Palmately compound leaf – In this type, all leaflets seem to be articulated at the tip of the
petiole, like fingers of a hand.
a. Multifoliate(digitate) – Having 5 or more terminal leaflets eg bamboo

Phyllotaxy
Phyllotaxy is the mode of arrangement or distribution of leaves on the stem and its branches
1. Opposite - when a pair of leaves arise from each node. If a pair of leaves arise at right
angles to each other, it is the opposite decussate.
2. Whorled - When three or more leaves arise from the same node.