VISAYAS STATE UNIVERSITY

COLLEGE OF ARTS AND SCIENCES

DEPARTMENT OF BIOLOGICAL SCIENCES
VISCA, BAYBAY CITY, LEYTE, 6521 PHILIPPINES

Name: Alastra, Vimson B. Instructor: Ris Menoel Modina

Course & Year: M. Ed-1 Bio Date: February 22, 2020
Schedule: Saturday (1:00-6:00) Score:

A Scientific Report on the Stages of Meiotic Prophase I of a Grasshopper

Using the Squash Technique

Introduction
Prophase meiotic stages have been noted in the recent studies since its stages are difficult to
interpret. It is also difficult to locate the stages of prophase meiotic cycle because of its lack on
chronological criteria.

To have a clear view and understanding on the different stages of meiotic prophase, a specie of a
grasshopper was used. The grasshopper was used to study the different meiotic stages in
Prophase I because of its availability, clear and large chromosomes, and easy to squash for the
beginners. An experiment is conducted to have a clear view on the meiotic stages of
spermatogenesis during Prophase I.

Materials and Methods

To observe the prophase meiotic stages, the reproductive gland that produces sperm of a matured
grasshopper is used. A matured grasshopper was collected, one (1) petri dish, one (1) dissecting
needle, five (5) glass slides, five (5) coverslips, one (1) 5 mL centrifuge tube, filter papers, one
(1) dissecting microscope, and one (1) light compound microscope. The reagents used for the
experiment was aceto-carmine stain, Carnoy’s fluid, 70% ethanol, chloroform, and acetic acid.

The experiment was done through squash technique using the testis of a matured grasshopper.
 A matured male grasshopper was collected

Small amount of chloroform was poured to slay

the grasshopper.

Small amount of 70% ETOH was poured in a

petri dish together with the grasshopper.

The testis of the grasshopper was removed

using a dissecting needle under dissecting
microscope.

Lobes of testis collected were placed in a 5-mL

centrifuge tube and washed with 75% ETOH.

The sample was soaked in aceto-carmine stain

for eight (8) hours.

Stained lobes were transfered in a petri dish

with 70% ETOH.

Samples were mounted in a glass slide with a

drop of 40% acetic acid and gently squashed
laterally.

Wiped off excess fluids and view the specimen

under light compound micrsocope (400x).

Results and Discussions

After preparing the sample slides of a grasshopper’s testis. The sample slide was placed and view
under High Powered Objective (400x) of light compound microscope. The process is repeated
several times until one (1) stage of meiotic prophase I is observed. The meiotic stage of Prophase
I that was observed under the light compound microscope was the Zygotene as shown in Figure
1.

Zygotene

Figure 1. Zygotene stage of a grasshopper’s testis viewed

under light compound microscope

The meiotic Prophase I has five (5) stages: leptotene, zygotene, pachytene, diplotene and
diakinesis. In leptotene stage condensation of chromosomes begins- chromosomes in this stage
cannot be distinguished. The second stage of meiotic prophase I is the zygotene. In this stage the
attachment of homologous chromosomes- the chromosomes begin to synapse. Synapsis of a
chromosome is cemented by a complex called synaptonemal complex, which facilitates crossing
over. Since in the zygotene stage the synaptomenal complex mediates the pairing of
chromosomes. When the pairing homologous chromosome is done, the crossing over takes place.
The crossing over between homologous chromosomes happens in the third stage of meiotic
prophase I- the Pachytene stage. Diplotene the fourth stage, here the condensation of
chromosomes continues and the homologous chromosomes starts to divide- disassembling of
synaptomenal complex. Diakinesis is the final period of 1st meiotic prophase. In this stage the
nuclear membrane starts to disappear. Prophase is known as the stage where nuclear envelop
disappear.
In the experiment conducted, only one stage of meiotic prophase is being observed due to some
reasons:
1) the collected grasshopper is not matured enough
2) the lobules or cells overlap
3) the sample is not fully squash

Those reasons leads to unsuccessful conduct of the experiment.

Summary and Conclusions
With regards of studying the different stages of meiotic prophase using the testis of a male
grasshopper. One of the best technique that is commonly used especially for the beginners is the
squash technique where the cell is pressed flat on a piece of glass and observe with the use of
microscope. In the experiment, a matured male grasshopper was collected. The testis of the
grasshopper have been removed with the use of dissecting needle under dissecting microscope
and soak with aceto-carmine for eight (8) hours. Making of sample slides using squash technique
follows. The prepared sample slide is viewed under a high powered objectives (400x) of light
compound microscope. The result of the experiment was not successful for some reasons. Of the
five stages of meiotic prophase, only one stage is being observed.

Recommendations
As mentioned, meiotic prophase is difficult to interpret because of its lack of chronological
criteria. The conducted experiment was not successful due to several reasons. To have a
successful experiment on the different stages of meiotic prophase one must consider the
following:

1) use a matured grasshopper;

2) when doing the squash technique, press or tap the coverslip gently to avoid
overlapping of cells; and
3) the sample must be fully squashed

References:
Biotechnology Methods / Cell Biology and Genetics
https://biocyclopedia.com/index/biotechnology_methods/cell_biology_and_genetics/meiosis_in_
grasshopper_testis_poecilocerus_pictus.php

Liu MH , Zhao KQ, Wang YD, Yang MP, Zhao NN, Yang DX

https://www.ncbi.nlm.nih.gov/pubmed/23262112

Schools of Biomedical Science WIKI

https://teaching.ncl.ac.uk/bms/wiki/index.php/Meiosis_prophase_1