EXPERIMENT SA

Preparation of a· stained temporary mount of an onion peel and to record observations and
draw their labelled diagrams.
~ORY: An onion is a multicellular plant organism. As in all plant cells, the cell of an onion peel
consists of a cell wall, cell membrane, cytoplasm, nucleus, and a large vacuole. The nucleus is 61
 pr om in en t, pr es en t in· th e ce nt re of the
m . Th e va cu ol e is
pr es en t atisthsue rrpe the [Link]
rip he ry ofcy
ou nd ed by toplasm
ce ll an d
 The presence of the cell wall and a large vacuole are indicators that identify plant cells, such
as cells in the onion peel.
[Link] S AND CHEMICAL S REomREn: A medium sized onion bulb, a glass slide, two watch
. glasses, a knife, a blade, distilled water, coverslips, forceps, a brush,: ne~dles, glycerine, a
dropper, safr~in solution, blotting paper, compound microscope.
PROCEDURE:

1. Take a medium sized onion bu~b. Using a knife, cut it laterally into
small pieces. Remove a leaf frOm one piece. _ •
2. Bend and gently break the above leaf frofil its middle. Carefully
peel the epidermis (covering) from
.
the inner surface
.
of the leaf.
.
. .. ,,.
. .

3. ~~ing-a forc~ps, remove the epidermal peel cind place it in a watch

glass which contains distilled-watef. .
.
4. Put 4-,5 drops of safranin-solution in _another watch gla~s. Using a
brush, lift the epidermal peel. from water, and place it in safranin
solution. Allow the peel .
to rest.. in -the . safranin. solution for at least
; '

3-5 minutes, so that the peel is properly stained. Be careful that·

the peel is not over-stained, nor is under-stained.
s. Using the brush, transfer the stained peel back to the watch glass.
containing distilled water. This is done to remove any extra stain
sticking
. to the peel. .
6. Take -a :clean, .dry glass slide. Using a dropper, put 2-3 drops fiJ
: epic
of dilute glycerine in the- middle of the slide. Glycerine is the
mounting material for the onion peel.
 7. Us ing a bru sh and a needle, tran_sf
er the stained-onion pee l to the mi dd le of the
ma y ten d to curl up. slid e. Th e peel
Flatten it gently, with the brush and needle.
8. Us ing bla de and needle, cut the pee
l to a square shape, approximately 2 mm or
3 mm .
9. Us ing a blo ttin g paper, gently rem
ove any. excess glycerine/stain around the pee
l..
1o. Pu t ano the r dro p of glycerine on .
the peel.
 Take a clean dry coverslip. Using a needle, lower it
gently on the peel in such a way that no air
• bubbles are anowed to enter. Using the brush and
peel so that the glycerine spreads over it evenly. . needle, gently flatten the coverslip over the
View the slide under a compound microscope.
E.R VAT lON ~•
0BSER\ATIONS:

1.A large n·umber of regularly shaped cells are lying side by side. Each cell has a distinct cell
wall.
2. A distinct (stained) nucleus is present on the periphery (one side) of each cell.
3. Lightly stained cytopla-sm is observed in each cell.
4. A large vacuole is present in the centre of each cell, surrounded by the cytoplasm.

CoNcLus10~: As cell walls and large vacuoles are clearly observed in all cells, the cells in
observation are plant cells.
 ... .-- ~~ - Cell membrane

- - ~ Nucleus

cell wall
_,_ __ _, Vacuoles
ce ll_ ..- --r 7
me mb rane nucleus

(a) (b)
( a) Structur e of onion peel un de r microscope (b) Structure of cells
• C ,\U 'f IO N S:
. As the p ee l is v e ry
apparatus to anotherd el ic at e, d o n o t hold it with •
. · •
. y~ur _~gers. Use a brus • • • •
Staining o f th e p e e l h to transfer it from• one
sh o u ld n ei th er b e to
Extra st ai n sh o u ld b o dark, nor too light.
e re m o v ed by· dipj,in
g the peel in water.
Hold th e g la ss sl id e
o n ly b y th e edges.
. .
The sp ec im en sh o u
ld b e m o u n te d iii th
e Ce~tre ·of the slide.
Extra g ly ce ri n e/ st ai • :' •
n /w at er sh o u ld b e re
Sufficient a m o u n t o moved; bj, u~ing biotti
f th e m o u n ti n g mat ng paper;
air b u b b le s in to th e erial, i.e. glycerine sh
co v er sl ip . ould be used to avoid
• • .• •. ·! ,·_: - : , the entry of
-•
• :: •• • •
 .. ,

Preparation of·a stained temporary- mount. of humari cheek: cells and to record observations
arid draw their labelled diagrams. • : •. • • • · •·

EORY: As in all animal cells, the cells of the' hurtian cheek do not possess' a cell Wall. A cell
(plasma) membrane, which is·semi-pei-meable; smtounds·.the cytoplasm.. Ulilike plant cells, the
cytoplasm in animal cells is denser, granular and occupies a larger space. The vacuole· in animal
 cells is smaller in size, or absent. The nucleus is pres ent at
the centre of the cytoplasm.
.

Abse nce o(._cell wall and prominent vacuole are indic ators
that identify animal cells; such as cells in the hum an chee k.
. .
..

APPA RATU S ·AND. CHEM ICAL S REQU IRED : A glass slide, a drop per,
a tooth pick , needles, a brush, methylene blue solu tion,
blott ing pape r, distilled Water, glycerine, a cove rslip , a
com poun d i;nicroscop(?.
 PROCEDURE:
Fig. 5.4 Structure of human
1. Take a clean glass slide. Using a dropper, put 1-2 drops of cheek cells
distilled water in the centre of the slide.
2. Rinse the mouth with fresh water and disinfectant solution.
3. ~ake a clean toothpick. Carefully scrape the inner side of your cheek to c0Uect some epithelial
tissue (cells). .
4. Put the tissue collected .on- the toothpick in the drops of water on the glass slide. Mix well.
5. Using a dropPer, put a drop of methylene ·blue solution on the above inixture on the glass slide.
Mix well and spread the ntjxture even~y .on the slide. Leave it undisturbed for 2-3 minutes, so
that the cell parts may get stained well. I •
• •

6. After 2-3 minutes, with blotting p·aper, carefully remove excess water and stain from the slide.
(Cace should be taken· NOT to remove the epithelial tissue as well.)
7. Using a dropper, put a drop of glycerine (mounting material) on the epithelial tissue.
8. Take a clean coverslip, and holding it with your left hand, lower-it carefully on the slide so
that it first comes in contact with the mounting material (glycerine). Then, using a needle,
slide it carefully over the specimen on the slide, making sure that no air bubbles enter. With a
brush and needle, gently flatten the cover~lip, ·so that glycerine is evenly distributed over the
.
specllllen.
9. With blotting paper, remove any extra liquid a~ound the coverslip.
10. View the slide under a CQmpo1:1nd microscope.

OBSERVATIONS:

1. A large number of flat and irregular-shape d cells are observed.

2. The cells do not have a cell wall. However, ~ach cell has a thin cell membrane (plasma
membrane).
3. A deeply stained nucleus (dark-blue stained due to methylene blue) is observed in the centre of
each cell.
4• No prominent vacuole is observed in the cells. The space between cell membrane and nucleus
is occupied by cytoplasm, which is granular in appearance.
s. '"fhere is no intercellular space b~tween the cells.
b th Ils of the
CoNcLus10N· As th
s . • e c~11s O served do not have cell wall, nor a prominent vacuole, e ce
pec1men on the slide are animal cells .

• ' •• \ 1 • • 4
I
a I • •
 -
Nucleus

f
l Cell
Membrane
-
. -.'
- ....
,
\
. . . . - ..... Jt.

- ._' .._,.·-
• '• l J

Nucleus Flat Polygonal Cells

Hum an cheek cells

 >

PRECAUTIONS:

I 1. Rinse the mouth with fresh wa ter and disinfecta

nt solution.
z. Ensure that the toothpick used to scrape the cheek is cle
an so it doe s not cau se inf ect ion to the
cheek. Also, scrape the ins ide of the che ek
gently so as not to cau se inju ry. ••
Also refer to "Pr eca utio ns" of Experiment~5A.
I

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