Scribd
Upload
4K views7 pages

BIO EXP 1 and 2

The document provides instructions for preparing temporary slides to observe pollen germination and the stages of mitosis in onion root tips under a microscope. Key steps include growing onion root tips in water, fixing and staining them, and mounting stained root tip sections on slides. Observations of interphase cells and the distinct stages of mitosis - prophase, metaphase, anaphase and telophase - are described, noting characteristics such as chromosome condensation and separation. Temporary slides allow for viewing the process of cell division in plant tissues.

Uploaded by

Bala Murugan.V
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
4K views7 pages

BIO EXP 1 and 2

The document provides instructions for preparing temporary slides to observe pollen germination and the stages of mitosis in onion root tips under a microscope. Key steps include growing onion root tips in water, fixing and staining them, and mounting stained root tip sections on slides. Observations of interphase cells and the distinct stages of mitosis - prophase, metaphase, anaphase and telophase - are described, noting characteristics such as chromosome condensation and separation. Temporary slides allow for viewing the process of cell division in plant tissues.

Uploaded by

Bala Murugan.V
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
You are on page 1/ 7

I SLIDE PREPARATIONS.

1.STUDY OF POLLEN GERMINATION

AIM: To prepare a temporary mount to study pollen germination on


the slide.
MATERIALS REQUIRED :

Fresh seasonal flowers, cavity slide, nutrient solution, brush,


needle, beaker, & Compound microscope.
PRINCIPLE :

(i) Pollen grain or microspore is the male gametophyte in


Angiosperm.
(ii) Pollen grains are usually spherical structure which measure
about 25-50 micrometers in diameter. In nature they germinate
on the compatible stigmas of the carpel.
(iii) In the laboratory their germination can be induced by placing
them in a pollen germination medium which contains nutrients
essential for pollen germination.
(iv) Each pollen grain is surrounded by a two layered wall referred to
as sporoderm.
(v) The outer layer of wall is called Exine and the inner layer is
called intine.
(vi) The exine is very hard and is deposited with Sporopollenin
which can withstand high temperatures and Strong acids and
alkali. No enzyme that degrades Sporopollenin is so far known.
(vii) Pollen grain has prominent apertures called “germ pores” where
sporopollenin is absent.
(viii) During germination, the intine of pollen grain emerges out as
pollen tube through one of the germ pores.
PROCEDURE:

(i) Dissolve 10g of sucrose in 100ml of distilled water to prepare the


nutrient medium.

(ii) Put few drops of the nutrient medium inside the cavity of the Slide.

(iii) Break the anther of a mature flower and dust the pollen grains in the
nutrient solution.

(iv) with the help of a needle place a cover slip on the slide and leave it
for about 10 minutes.

(v) Observe the slide first under the low power of the microscope to see
pollen germination and then under high power of the microscope.
OBSERVATIONS:

Numerous pollen grains put forth pollen tubes in the nutrient


medium.

RESULT :

Pollen grains germinate in the nutrient medium.


PRECAUTIONS:

(i) use only freshly plucked flowers

(ii) The cavity slide and cover slip should be clean.


2.STUDY OF MITOSIS:

Aim: To prepare a temporary mount to study the different stages of mitosis


in onion root tips.

Materials Required:

Onion bulb, wide mouth glass jar/bottle, glacial acetic acid, ethanol,2-
4% acetocarmine stain, N/10 HCI, spirit lamp, slide, cover slip,
blotting paper, needle, blade, and compound microscope.

PRINCIPLE

During somatic growth in the body, number of cells increases. The


process of cell division is responsible for the increase in number of
cells in an organism. A somatic cell divides by mitotic division to form
two daughter cells. The characteristic feature of mitotic division which
differentiates it from meiotic division is that in the daughter cells the
number of chromosomes remains the same as in the parent cell. In
plants, mitotic division takes place in meristematic tissues which are
located in the apices of root and shoot. So one can easily observe
and study different stages of mitosis by preparing temporary mounts
of apices of root and shoot.

PROCEDURE:

(a) Growing of Root Tips

1. Take a medium-sized onion bulb and remove the dry roots from
its base.

2. Place bulb on the mouth of a glass bottle/jar filled with water in such
a way that the stem portion of the bulb (basal part) just touches the
water.

3. Within 3-6 days, new roots will appear from the bulb. When roots are
2-3 cm long, cut them from the tip.

4. Transfer the root tips into a vial containing freshly prepared mixture of
1 : 3 glacial acetic acid and ethanol. This will fix the root tips. In onion,
usually cell division takes place about two hours after sunrise.
Therefore, roots grown on water should be cut and fix only at that
time.

5. After 24 hours, transfer the root tips to 70% ethanol (for preservation
and use in future).

Preparation of Slide

1. Take one or two preserved root tips and wash them in water on a clean slide.

2. Place one drop of N/10 HC1 on the root tips followed by 2-3 drops of acetocarmine solution.
Now warm the slide slightly on spirit lamp. Care should be taken that the stain is not dried up.
Using a blotting paper carefully blot the excess stain.

3. Now with the help of a blade cut about 2-3 mm tip portion of the root which is comparatively
more stained. Retain the comparatively more stained portion on the slide and discard the
remaining portion.

4. Put one or two drops of water on the stained root tip and blot it carefully using blotting paper.
Again put a drop of water on the root tip and mount a cover slip on it avoiding air bubbles.

5. To squash the meristematic tissues of the root tip, slowly tap the cover slip using the blunt end
of a pencil or a needle. As a result of this, the meristematic tissue will spread in the form of a
thin layer of cells.

6. Place the slide on the stage of a compound microscope and first observe it under the lower
magnification to locate the area having a few dividing cells. To study the detailed features of
mitosis, examine the dividing cells under higher magnification.

OBSERVATIONS:

 Under lower magnification rectangular cells with pink nucleus


are observed.
 Under higher magnification most of the cells observed in a
microscope field are not in the phases of cell division. These
cells are considered to be in interphase. Cells showing mitotic
division are in different phases. Some of the characteristics of
interphase and each phase of mitosis are as follows:

Interphase:
1. It is the non-dividing phase of the cell cycle which occurs between
two successive cell divisions.

2. The cells are rectangular in shape.+

3. Each cell contains a densely stained large nucleus almost in the


centre.

4. The coloured material of the nucleus is homogeneous and appears


granular.

5. The nucleus has a distinct boundary and one or few nucleoli.

Stages of Mitosis

(a) Prophase

1. The nuclear membrane starts to disappear and at the late prophase it


disappears completely.

2. In the early stage, nucleolus is visible but in later stage, it disappears.

3. The chromatin opens up and chromosomes become clear. In the


early stage, chromosomes are very thin but at late prophase, they
become comparatively thicker and rod-like.

(b) Metaphase

1. The nuclear membrane is absent.

2. The nucleolus is not observed during this phase of cell division.

3. Thick and rod-shaped chromosomes are seen arranged at the


equatorial plane of the cell.

4. Each chromosome at this stage has two chromatids joined


together at the centromere.

5. Fine fibrils form spindle-shaped body. Two ends of spindle are


called poles.
6. Centromere of each chromosome is attached to fine fibrils of
spindle-shaped body.

(c) Anaphase.

1. In this phase, the spindle fibres start to shorten, as a result two


chromatids of a chromosome which are still attached to centromere
start to separate.

2. The centromere splits and now each chromatid represents a


separate chromosome and it has its own centromere.

3. The chromosomes drag towards the opposite poles of the spindle.

4. The chromosomes at this stage assume the shape of inverted ‘V’,


‘J’ or ‘I’ depending upon the position of centromere in them.

(d) Telophase and Cytokinesis

1. Chromosomes reach the opposite poles.

2. Chromosomes become thin and they entangled to form a mass


called chromatin.

3. Nuclear membrane and nucleoli reappear to form the nuclei of the


two future daughter cells.

4. In plant cell, after telophase a cell plate is formed in the middle.


This cell plate extends towards the periphery of the cell. As a result,
the parent cell divides into two daughter cells.

PRECAUTIONS :

1. The base of the onion should be in contact with water while


growing the roots.

2. The root tips should be cut and fixed early in the morning.

3. Slide and cover slip should be clean.

4. Slide should be warmed gently much above the flame of burner.


5. There should be no air bubble under the coverslip.

You might also like

pFad - Phonifier reborn

Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.

Note: This service is not intended for secure transactions such as banking, social media, email, or purchasing. Use at your own risk. We assume no liability whatsoever for broken pages.


Alternative Proxies:

Alternative Proxy

pFad Proxy

pFad v3 Proxy

pFad v4 Proxy