EXPERIMENT-1

Aim of the experiment.

To study the different parts of a compound microscope.

MATERIAL REQUIRED

 Compound microscope
 Permanent slide

Theory
A compound microscope is high magnification microscope that uses two lenses to achieve
the real and magnified images of objects and a high magnification microscope that uses
two lenses to achieve the real and magnified images of objects and helps multiply the
magnification level. The first lens is referred to as the objective lens and typically has a 4x,
10x, 40x or 100x magnification ability. The second lens is known as the eyepiece lens.

Parts of Compound Microscope

The parts of the compound microscope can be categorized into:

 Mechanical parts
 Optical parts

Mechanical Parts of a Compound Microscope

 Arm
o The whole microscope is handled or carried by the curve-shaped
structure called the arm.
 Body tube
o The body tube separates the objective and the eyepiece of microscope.
The body tube can be shifted down and up using the adjustment knobs.
 Foot or base
o It is a U-shaped structure and supports the entire weight of the
compound microscope.
 Stage
o The flat and rectangular plate that is connected to the arm’s lower end is
called the stage. It is the platform upon which the specimen or slide are
placed for studying and examining the various features. The height of
the mechanical stage is adjustable on most compound microscopes. It
has Stage clips on the stage that hold the slide in place on the
mechanical stage. The centre of the stage has a hole through which light
can pass.
 Clips
 o The upper part of the stage is connected to two clips. These clips on the
stage used to hold the slide in its position.
 Diaphragm
o The diaphragm is found under the stage of the microscope and its
primary role is to control the amount of light that reaches the specimen.
It’s an adjustable apparatus, hence controlling the light intensity and the
size of the beam of light that gets to the specimen.
o The diaphragm can be of two types:
 Disc diaphragm
 Iris diaphragm
 Nose piece
o The nose piece is circular and a rotating metal part that is connected to
the body tube’s lower end. The nose piece has three holes wherein the
objective lenses are embedded.
 Fine adjustment knob
o It is the smaller knob, which is used for sharp and fine focusing of the
object. For accurate and sharp focusing, this knob can be used.
 Coarse adjustment knob
o It is a large knob that is used for moving the body tube down and up for
bringing the object to be examined under exact focus.
 Light Source
o The light source in the microscope is a lamp that you turn on and off
using a switch. You can adjust the intensity of light by turning the light
adjustment knob.

Optical Parts of Compound Microscope

 Eyepiece lens or Ocular

o At the top of the body tube, a lens is planted which is known as the
eyepiece. On the rim of the eyepiece, there are certain markings such as
4X, 10X, 40X and 100 X. These indicate the magnification power. The
object’s magnified image can be observed with the help of an eyepiece.
 Condenser
o A condenser sits between the stage and the diaphragm. The condenser
controls how much light from the illuminator is permitted to pass up
through the aperture, it controls the intensity of the light.
 Objective lenses
o The objective lens gathers light from the specimen, magnifies the image
of the specimen, and projects the magnified image which can be
observed through eyepiece. There are three objective lenses are as
follows:
 Oil immersion objective – 100X
  High power objective – 45X
 Low power objective – 10X

Procedure

 Place the microscope on the working table and remove dust by wiping the body
with a silk cloth.
 Clean the lenses with lens cleaning fluid and lens cleaning paper. Identify the
various parts of the microscope.
 Draw a diagram of the microscope and label its various parts.
 Take a permanent slide preparation or a temporary preparation made by you, and
keep it on the stage of a microscope, fix it with clips after focusing and view.
 Learn how the microscope can be tilted or inclined by moving the arm of the
microscope.
 Note, how focussing is done by moving the coarse adjustment and fine adjustment
knobs.

Precautions

 The objective lenses and eyepiece should be cleaned with the help of silk cloth
and cleaning liquid before use.
 The microscope should not be tilted when working, using it.
 When an object needs to be studied, focus on the low power objective first and
then move to high power.
 When focusing, care needs to be taken to ensure that the objective lens never
strikes the stage or the slide.
 Only the fine adjustment knob should be used when the high-power objective is
employed.
 Coverslip should always be used to cover well-mount preparations before
observation under the microscope is made.
 Do not dismantle the microscope.
 An oil immersion lens should never be used without the use of oil.
EXPERIMENT-2

Aim of experiment

To study the stomatal distribution on the upper and lower leaf surfaces and to calculate the
stomatal index.

Materials required

(i)Glycerine

(ii)Safranin solution

(iii)Forceps

(iv)Leaf

(v)compound microscope

(vi)Needle and brush

(vii)Cover slip
(viii)Dropper

(ix)Blade

(x)Watch glass

Theory
Stomata are minute pores found on the epidermis of leaves and young shoots of plants
that are used to control exchange of gases. The pore is surrounded by a pair of specialised
cells called the guard cells that are responsible in regulating the size of the opening.

Water is released through the stomata into the atmosphere in the form of water vapour
through the process called transpiration. Besides this, the exchange of oxygen and carbon
dioxide in the leaf also occurs through the stomata.

Distribution of Stomata
Distribution of stomata varies between monocots and dicots, between plant species, and
between the underside and top side of the leaves on a plant.

Stomata are found more on plant surfaces thriving under higher light, lower atmospheric
carbon dioxide concentrations and in moist environments.

Usually the lower surface of a dicot leaf has a greater number of stomata while in a
monocot leaf they are more or less equal on both surfaces. In most of the floating plants,
stomata are found only on the upper epidermis.

Procedure

 Pluck one fresh leaf of a plant.

 Take two watch glasses and pour some distilled water into the both watch glasses.
 Split the leaf from the plant obliquely.
 Take the peel from the upper surface of the leaf using the forceps.
 Place the peel into a watch glass containing water.
 Take another peel from the lower surface of the leaf using the forceps.
 Place the peel into the other watch glass containing water.
 Using a dropper, take few drops of Safranin solution and put it into the two watch
glasses.
 Take two clean glass slides and place the leaf peel on the slides one by one, using a
brush.
 Take a blade and cut a small rectangle or square piece from each peel.
 Take some glycerine using a dropper and put one drop of glycerine on both slides.
 Take a cover slip and place it gently on the peel with the help of a needle.
 Take the glass slide and place it under compound microscope.
 Observe under the microscope.
 Count the number of stomata in the peels of both upper and lower epidermis of the
leaf appearing in the microscopic field.

 Calculation of Stomatal Index

  The distribution of stomata on the upper and lower surfaces of the leaf can be
studied by removing the peels of the leaf from the upper and lower surfaces and
observing the same under a microscope.
 The count of the the number of stomata and epidermal cells in the microscopic field
is taken and the stomatal index of each surface of the leaf can be calculated using
the following formula:

EXPERIMENT-3

Aim of the experiment

To prepare a temporary stained mount of a transverse section of dicot and monocot stem to
study various plant tissues.

Material Required
 Preserved material stem
 Microscope
 Sharp blade
 Slides
 Watch glass
 Coverslips
 Safranin (1gm in 100ml of 50% ethanol)
 Brush
 Glycerine
  Blotting paper

Procedure

Taking Sections
 Hold the dissected plant material between your index finger and thumb, while keeping
the edge of the razor perpendicular to the longitudinal axis of the plant. Slice it into
thin sections.
 Using the edge of the blade, shift these sections into a watch glass with the help of a
brush. The watch glass must hold water.

Process Of Staining
 Pick 2 to 4 thin and good transverse sections. Shift it to a different watch glass
holding safranin stain.
 Let the complete set rest in the stain for a couple of minutes.
 After a while, drain the sections of the stain and rinse again with water so as to wash
off the excess strain.

Mounting
 On a clean slide, place a stained section in the middle of the slide, mounting water or
glycerine.
 Using a needle slowly places the coverslip.
 An excess of water or glycerine from the edge of the coverslip can be removed by
placing blotting paper.
 Ensure, that the air bubbles are not formed while mounting.

Aim
To study the transverse section of a dicot stem, a sunflower stem.

Theory
 A typical example of a dicot stem is a sunflower
 The outline of the stem in the transverse section is circular with a hairy surface
 The transverse section depicts the arrangement of different tissues in a particular way
from the exterior to the centre as listed below:
  Epidermis

 It is the outermost covering of the stem and is single-layered

 The cells are living cells that are densely arranged with a thin wall,
enclosed in a cuticle

 It displays multicellular hair

 Function – It provides protection

 Cortex

 Located just under the epidermis

 It consists of the following regions – Hypodermis, Endodermis,

General cortex.
(a) Hypodermis – It is situated just below the epidermis and consists of 4 to 5 layers of
collenchymatous cells. These are cells that have the cellulose deposits on the edges and are
living cells. It may possess chloroplast and hence carry out photosynthesis.

Function: It renders mechanical support

(b) Endodermis – Forms the interior most covering of the cortex consisting of a singular row
of cells that are densely arranged and barrel-shaped with no intercellular spaces. They are
composed of starch grains.

(c) General Cortex (Parenchyma) – Located just under the hypodermis, it consists of
several layers of cells that are living having thin walls along with intercellular spaces. It may
contain chloroplast and hence becomes photosynthetic. Several mucilaginous canals can also
be observed. Function: Food storage

1.

 Stele
In tissue, it forms the central core having a vascular bundle. It comprises of – pericycle,
vascular bundles, medullary rays, and pith.

(a) Pericycle

 Located between the vascular bundles and endodermis.

 Has recurrent patches of parenchyma and sclerenchyma.

 Each sclerenchyma patch is linked with hard-blast fires.

(b) Medullary rays

 It is located in the gaps between the vascular bundles. This gap finds
parenchymatous cells that are thinly walled and arranged radially in 4-
5 rows.

 Helps in the storage of food material and in the lateral conduction of

water and food.
(c) Vascular bundle

 They are open, collateral and conjoint and are set up in a ring-like
manner.

 Each of the vascular bundle comprises of- xylem, phloem, cambium.

 Phloem is located below the pericycle towards the exterior side. It

consists of thin cell walls possessing companion cells, sieve tube and
phloem parenchyma and conducts food material.

 Xylem is found in the interior of the vascular bundle, i.e., smaller

protoxylem towards the centre and larger metaxylem towards the
periphery. Therefore xylem is endarch. These cells are dead and
lignified. Xylem conducts minerals and water.

 The cambium is present between phloem and xylem. The cells are
rectangular and have thin cell walls. It consists of meristematic tissues
that generate new cells.
(d) Pith

 It forms the mid-region of the stem. It arises under the vascular bundle
up till the centre and consists of large parenchymatous cell

Identification
 The epidermis has multicellular hairs.
 The hypodermis is collenchymatous.
 The xylem is endarch with the protoxylem being at the centre and the metaxylem
being at the periphery.
 Vascular bundles are open, collateral and conjoint.
 They are arranged in a ring-like manner.
 Pith is found at the centre.

Conclusion
The specimen provided is a section of a dicot stem.
 T.S of monocot stem – Maize stem

Conclusion
The specimen provided is a section of the monocot stem