Cell biology/Meiosis/Lab

Procedure for studing meiosis stages AIM: To observe and study meiosis stages.

MATERIALS REQUIRED: a mature male grassgopper ,embryo cup-1,gass slide-1 ,coverslip-1,needle -1,brush-1,Carnoy's solution,100% alcohol ,90% alcohol ,70% alcohol,distill water ,Ferrous ammonium sulphate, Haematoxylin stain, 45% of acetic acid,pencil ,compund microscope

1.Collect a mature adult male Poekilocerus pictus (grasshopper).

As it feed on Calatropis gigentia ther are high chances you can collect from here.

To distinguish and identify between male and female grasshopper following easy keys you can follow:

a.Female abdomen is broader than male grasshopper.

b.Female is brownish in colour and male is green in colour.

c.Female abdomen is bifid than male abdomen.

2.Take out two pair of testis by squezing the male grasshopper from neck to the abdomen.

Thus you have pair of testis having millions of testis follicles.

3.Keep follicles in embro cup having carnoys fluid.

Carnoy's solution is a fixative composed of 60% ethanol, 30% chloroform and 10% glacial acetic acid.

4.Separate the follicles from other tissues .

So many follicles clustered together give appearance like aggregation of berry.

Each follicle is small drop shape white stucture.

(always transfer material with the help of smooth brush)

5.Transfer follicles in 100% alcohol for five minutes kept in embryo cup.

6.After five minutes, transfer follicle into 90% alcohol kept in embryo cup.

7.After 5 minutes ,tranfer follicles into 70% alcohol.

You can store them for months together.

8.Take out 2-3 folliles from cluster and keep them in embryo cup having distill water for five minutes.

Take Ferrous ammonium sulphate in embryo cup.

9.Transfer follicle in ferrous ammonium sulphate and leave it undisturbed 10 minutes.

10.Again transfer them in distill water for 5 minutes.

11.Take Haematoxylin stain in embyo cup .

12.Transfer follicle into haematoxylin and keep them undisturbed for half an hour.

13.Tranfer one follicle onto the clean glass slide with the help of brush.

14.Chop into small pieces and take only point material for avoid crowding and better sqaushing.

15.Put one drop of 45% of acetic acid on to the slide .

16.Put a clean coverslip on the material without avoiding air bubbles.

17.Squash the material by gently tapping on the coverslip by blunt end of pencil keeping blotting paper on it.

18.Focus the slide under compund microscope and locate the meiosis stages.

19.Immediately after identification seal the slide with wax.

20.DRAW DIAGRAM OF STAGES IN YOUR NOTEBOOK.

OBSERVATION:DIFFERNT STAGES OF MEIOSIS 