VIll:

Practical
Microolology
Unit

DROP
EPARATION
P R E P A R A T I O

HANGING
motilit of bacta
observethe
IL. usedto bacterial ggrowth
r o w t h provided in
dropis the
b a c t e r i a l

( H a n g i n g
from tube
test
in a
prepared
of bacteria

It is
m e d i u m

culture
pared from

liquid
ed
a n dt h e n

Method(Fig 54.3) to you I place

proVIded

ring
of
glass
slide (Step ).
plasticin

Make a clean
1.
the centre
ofa bacterial
culture put it and

it in liquid
of 2).
with the help
2. Take
a loopful cleancoverslip (Step
centreofa c o n t a i n i n g the
asticin ring
plasti
in The de
siIde
the bacterial

3. Invert the
glass
onthe
cOversli (Step 2) in
a place 1tsticks with the
make
and
er to
(Step 1) the coversl1p
so that
such a manney' the
c e n t r e of the
remaining in
the smearfor
plasticin and thedrop
plasticin ring(Step 3). coverslip_into
ming the side slide along with
the glass
4. Bring back
the smear. position(Step 4). coversl1p, thats
theoriginal beneath the
hangs
5. The drop now
called hanging drop,
why it is the microscope
preparation under
crystal violet
Examine the
te (Other dyes 6.
be (Refer below).
letmay also
for one minute.

tone for 10-30

olourise.(Alcohol

to remove the

a plasticin ring
Step 1-Glass slide containing
nin (counterstain)
hsin or neutral red

X100) objective. medium

Step 2- Coverslip with a drop of culture
TO
0.5 gm
100ml

Step 3-Inveptéd glass slide from

gm step 1 placed on to coverslip in step 2
2ggm
100 ml

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P a c t i c a l M i c r o b i o l o y

vsualsed under
examiehe as
Hanging ote
visualsed under
pepeeion as
Hanging coo
XO obecthve
hanging drop under the
microscopN

Fig 544 Examination of

Method (Fig. 545)
as
are prepured
of stool speimen
Wet preparations
PreoautioS ign-hick or very of
plasticin_ing follows preparation

the as iodine
0 Do not make be of appropriatethicknes drop 1Saine
aswell
glass shide
preparation

thin.It should
same
the hanging made on the put at on
focusSing stool are cent) is
ditheulty in is used lt normal saline
(09 per
) There is plasticin ring A drop of the
end of other
thick obiective 2 while on

preparation when
slide
coversip while end of a
clean glass pat solution is
breakage of of iodine the
might resultin shde, a dop both in
focussing added same
loweredfor of facces is
the
dop to make
enss being plasticine ring.
the hanging
exminatian
3A
minute portion
of a small
stick wooden

of thin slideand with the help

(12) In case touches the
glass drops
cach
a unifornm suspension
suspension

over
preparation placed no
ditticult.) that
becomes is gently taking
care

ofdrop 4 A covershp
iodine)
separately,
and
Examination (Fig. 54.4) (saline objective

hanging drop air bubble forms low power

edge of a
under
the (X10) ot preparation

1Always fist focus low power objective Examine

the
(see
below)

microscope

preparation
,under lens of
and

mieroseope objective (X40) that

pouer
föcus to high
sure
Make
T u r n the motileyof
baeteria.
also be
visible

observefor the
sheuld
hanging drop
the
theedgeof
under high power.

(NI00) for
Precautionms objective
immersion Nwmmel s a l n e

Never use_oil
preparation

0 hangingdrop examinationJ
bacteria under the
Neyerat

545
Stool

motility of Fig.
look for the not under the low e o h y s

n
Always
(X40)and
A v l e

or
objective saline

npower preparation if to
ether oy
poiwer objective (X10. P r e c a u t i o s

Ie
nmivw
drop pr fase adding
laeces
should

(11) Never xamine the hanging

preparation
may
show
non ()
Before container

Sometimes
ovaor
m
up/Dry n
as
seen
be
has driedmotle bacteria may
a c e s

mixing
stick.
the the
to repeat
Wooden c o n t a n e r

Tesults ie always
better
the within

Therefore, it is interpretation
d o w n

of
in these
us
hotile. for proper distribution driol
preparation has
nging drop preparation
fore x a m i n a t i o np u r o w e s

1 . STOOL/FAECES EXAMINATION

(
ot
cysts
and
to find out on
examination
is done
different par Sites
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