Cytology Project in 18 page

7/23/2019 Cytology Project in 18 page

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INDEXS.n

oChapter Page No.

CYTOLOGY STAINING

1 Introduction 3

2 Clinical Significance 4

3

Sampling Techniques in cytology

1. !"foliation of cells noa#rasi$e

methods2. Collections of cells remo$ed #y

#rushing or similar a#rasi$e

technique3. %ashing or la$age4. &ine needle aspiration cytology

'&N(C)

*

4 Preparation of smear +

* &i"ation +

+

Types of stains ,

1. Pap-s stain2. omano/s0y stain3. Cresyl $iolet stain4. epid stain*. Special stain+. emato"ylin !osin stain. Shorr-s stain

to 1*

aintenance of the stains 1+

5 Consolation 1

6 eference 15

17 (nne"ure


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1. INTERODUCTION8 Cytology is a #ranch of diagnostic

medium it deals /ith study of free cells from shade from

#ody. Itself or aspirated #y tu#e of needles9 therefore

effusion secretion9 aspirates scraping all are use in

diagnostic cytology.

It can #e referred to as cell #iology or cytopathology

according to the specific use of cytology. Cytopathology is a

#ranch of pathology that steadies and diagnoses diseases on

the cellular le$el.


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2. CLINICAL SIGNIFICANCE8 Cytopathology is usually

used to aid in the diagnosis of cancer. o/e$er it also

help in the diagnosis of certain infection discuses and

other inflammatory conditions. It is also commonly used to

in$estigate thyroid lesion9 disease in$ol$ing slerile #ody

ca$ities.

%hich studies /hole tissues a common application of

Cytopathology is the :pap smear; used as a screening tool9 to

detect precancerous cer$ical lesion and pre$ent cer$ical

cancer.


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3. SAMPLING TECHNIQUES IN CYTOLOGY8

Exfoliaio! of "#ll$ !o!a%&a$i'# M#(o)$*+ This is

spontaneous shedding of cells deri$ed from the linling of

an organ into the #ody ca$ity. &or e"ample $aginal

smears9 sputum9 urine and other #ody fluids.

Coll#"io! of "#ll$ ,o'#) %- %&$(i!/ o& $i,ila&

a%&a$i'# #"(!i0#*+ The method allo/s direct samplingof the targets such. (s surface of the uterine cer$i" or

#ronchus. The sampling of organs of the respiratory

gastrointestinal tract is possi#le

a$(i!/ o& la'a/#*+ Normal saline or similar solution are

installed in the target organ under $isual control.

This solution are aspirated collected in small

container

Fi!# !##)l# a$i&aio! "-olo/- FNAC4*+ (ny palpa#le

or non palpa#le lesion in the #ody can #e sample suing

either palpitation or imaging techniques . the technique

can #e adopted as an outpatient procedure. &N(C is raip

cast effecti$e methodology.


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5. PREPARATION OF SMEAR*+ Smear are made

immediately form s/a# #y8 Smear should #y thin e$enly spread /ithout clump.


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7. TYPES OF STAINS8

a. Pap-s stain 'papinoculo stain)#. omano s0y stainc. Cresyl $iolet staind. apid stainse. emato"ylin !osin Stainsf. Shorr-s staining

Pa8$ $ai! ai!o"lo $ai!4*+ The most commonly

used stain in the qynac cytology is the papinoclo stain the

important of these stain is suita#le for cer$ical smear.

M#(o)*+ This staining ethod can #e perform either

manually or automatic.

Solio!8

a. erry-s emato"line.

#. +c. !osin (nure*7

P&o"#)

a. emo$e smear from fi"ati$e.#. inse in descending grade of alcohol 57?9 7? *7?

and /ater for 17sec. it.c. Stain in erry-s emato"line for 2 minutes.d. inse in /ater for 2 minutes.e. @ifferentiate in 1? acid alcohol until only the nuclear retain

the stain.f. %ash Alue in ta# /ater for 3* minutes.g. Transfer to 7? alcohol for fe/ seconds.h. Transfer to 6*? alcohol for a fe/ seconds.i. Stain in organ >+ for 2minuts.

B. inse in 2 changes 6*? alcohol.0. Stain in eosin (ure , *7 for 24 minutes.


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l. inse in 2 changes 6*? (lcohol.m. @ehydrate in (#solute (lcohol clear in "ylene and ount

in natural synthetic modicum.

R#$l8

Nuclei Alue

(cidophilic Superficial cell

Aasophilic Alue

Canada ed pale pin0 Tricomonus Deginitis >rayish >reen


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Ro,a!o9$:- Sai! 8 &ollo/ing &i"ation in either alcohol

smears may #e any of the omano/s0y stain.

The most popular omano/s0y stains are the >iemsa

and the =eishman stain.

M#(-l#!# ;l#*+ This signal stain rapid simple

method is useful for the screening of fresh specimens

especially sputum for malignant cells.

Solio!*+

ethylene 1 /,@istilled /ater 1


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CRESYL =IOLETSIN*+

I,o&a!"#*+ any la#oratories use this method since it

is simple fast and gi$es clear morphologic cellular details. R#a/#!$*+

7.1 gl '$E$) cresyl $iolet

1? acetic acid

P&o"#)*+

Prepare the smear and fi" in ether alcohol fi"ati$e

ydrate the smear in the follo/ingo 7 ? '$E$)

o *7 ? '$E$) ethanol

o @istilled /ater

@ip in 1?'$E$) acetic acid for 37 sec.

inse in distilled /ater '2 changes)

Stain in freshly filered cresyl $iolet solution for 3 min.

@rain and dehydrates #y rinsing in acetone '#rieflyfor 17 and * seconds respecti$ely)

Clear in "ylene for * minutes each

ount in Canada #alsam '@PF)

A))iio!al i!fo&,aio!*+


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arris hemato"lin

1.7? '$E$) hydrochloric acid in 7? ethonal 'acid

alcohol reagat) !osin 'y) 8 1? '/E$) aqueous.

P&o"#)*+

Treat /ith 6*? alcohol for 2 min.

%ash in running tap /ater for 23 min.

Stain in arris hemato"ylin for *17 min.

%ash in running tap /ater for * min.

@ifferentiate in acid alcohol solution for *17 min.

%ash in running tap /ater for *17 min.

Treat /ith eosin solution using 2 dips

Alat and dehydrate in a#solute alcohol

Clear in "ylene and fi" co$er slip #y using a drop of

@PF

R#$l *+

Nuclei 8 Alue

Cytoplasm 8 Pin0


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Rai) Sai!$*+ These stains are used mainly to chec0

adequacy of fine needle aspirates9 apid stein can also #e

used for immediate e$olution to control cross contamination.

The dyes used in rapid steaming procedures are ethylen

#lue. Thionin #lue Toluieline #lue.

Toi)i!# %l# $ai! i$ a) a$ follo9$8

Toluidine #lue 8 7.* g

6*? ethyl alcohol 8 27 ml

@istilled /ater 8 57 ml

@issol$e dye in alcohol ass /ater filter store in dar0

#ottle.

Toli)i!# %l# $ai!i!/ &o"#)*+ It is performed as

follo/s.

&i" the smear for 17 to 1* seconds in 6*?

alcohols.

Treat /ith toluidine #lue use 17 dips

inse in /ater to remo$e e"cess of stain.

Put a co$er slip on /et smear su#mit for

microscopic e"amination


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S#"ial Sai!*+ These are /idely use in histopathology la# can

#e applied to cytological preparation.

In cytological e"amination /hich is use for thein$estigation called ID created in$es #ronchial al$elar

la$age for pneumocytis is carinei.

This is used to quite easu to ma0e a diagnosis of

>> papinoculo stained smear.

Solio!*+

1. M#(-la,i!#*+ Sil$er nitrate 'Stoc0 solution and their

/or0ing solution) *? of chromium trio"yle 1? of sodium #isuffite 7.2? of >old chloride 12? of sodium thyosulfate

2. P&o"#)*+

a. %ash slide in @.% '1727 dips)#. Transfer to cooplin for /ith *? chromium trio"ide

place 1ur in a /arn /ater 43*57c 27 minc. inse in tap /ater 1727 dipsd. Transfer to sodium #isulphate 37sece. unning tap sodium #isulphate 37 secf. unning tap /ater 1*27 secg. Se$eral rinse of @.%

h. !thylamine sil$er main /ateri. inse in @.% 3 times 17 to 1* days each

B. 7.2? >old chloride 2737 dip0. inse in @.% 3 times 171* dip eachl. 2? Sodium thyorulfate for 1 minm.unning tap /ater 1*27 secn. &ast grain 373* seco. inse in @.% l/ice 1*27 dips

p. 6*? ethyle alcohol t/ice 1*27 dipq. (#solute alcohol t/ice 1*727 dips


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r. Fylene 'Clearing)s. ount

R#$l*+ Pn/mocytisis corinic fungal element Alac0

usine glycogen pale gray

Aac0 ground pale green


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S(o&&8$ Sai!i!/*+ This method uses a single differential

staining solution. It gi$es less cytoplasmic transparency and

poorer nuclear definition than the papanicolou.

It is not $ery useful in the diagnosis of malignant

cells.

Sai!i!/ Solio!8

*7 ? ethyl alcohol 177 ml

Aie#rich Scarlet 'ClNo. 2+67*)

%ater , Solu#le 7.* g

'Cl No. 1+237) 7.2*g

&astgreen &C& 'Cl no. 427*3)


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?. Mai!ai!#"# of (# $ai!$8 &ollo/ing precautions are

ta0en /hile using the steins8a. Stain used must #e certified #y Aiological stein

commission.#. @ye content on the certified la#el of dye must #e

considered in stein preparationc. Contamination control >ynec Nongynec slides are

stained separatelyd. &itter stains reagents dailye. Non>ynec Specimens ha$e high potential for cross

contamination during staining procedures.f. Solutions and stains are changed from time to time

depending on the $olume of /or0g. Staining dishes should #e 0ept co$ered

h. (lcohol used during rehydrating and dehydrating

process are changed on a rotation #asis and if needed

also filtered prior to staining procedurei. @aily microscopic chec0 are documented for the quality

control of stains.


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@. CONCLUSION*+

The focused ultrasound therapy is safe

and effecti$e ethod for treatment of chronic

cer$icitis and is /or0ing to #e popularied.


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. REFRENCE*+1. Te"t #oo0 of edical la#oratory Technology

3rd9 Praful A.>od0ar9 @arshan P.>od0ar9

Ahalni pu#lishing ouse9 um#ai9 27142. edical =a#oratory Science9 G. ra/ !ducation pri$ate

=imited Ne/ @elhi2717

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Cytology Project in 18 page