CHAPTER
35
Immunocytochemistry
Mamatha Chivukula and David J Dabbs
Contents
In tro d u c tio n
B re a st C y t o lo g y
Im m u n o c y to lo g y T e c h n iq u e s
E -c a d h e rin a n d p 1 2 0 C a te n in
T h e S p e c im e n
Im m u n o c y to c h e m is try fo r T a r g e t e d T h e ra p ie s
In te rp re ta tio n a n d L im ita tio n s o f IC C
C D 1 1 7
E ffu s io n C y t o lo g y
H E R 2 /n e u
M e so th e lia l M a rk e rs
G y n e c o lo g ic C y t o lo g y
C a lr e tin in
p i 6 ink4a
H B M E -1
P ro E x C
C y t o k e r a t in - 5 /6
In S itu H y b r id iz a t io n A s s a y s f o r D e t e c t io n o f H ig h -R is k H P V
W ilm s ' T u m o r G e n e P r o d u c t
O v a ria n C y t o lo g y
D 2 -4 0
\
A
/’
1
~T
n
1
I
G L U T -1
N o n M e so th e lia l (A d e n o c a r c in o m a ) M a rk e rs
M O C -3 1
C a rc in o m a o f U n k n o w n P rim a ry
B G -8
B e r-E P 4
N o n e p ith e lia l M a lig n a n c ie s P re s e n t in g a s T u m o rs
M o n o c lo n a l C E A
o f U n k n o w n O rig in
T A G -7 2 ,3
M a lig n a n t M e la n o m a
C D 1 5 (L e u -M 1 )
K i6 7
S ite -s p e c ific M a rk e rs
c - m e t
T h y r o id T r a n s c r ip tio n F a c to r-1
C o n c lu d in g R e m a rk s
E s tr o g e n R e c e p to r
C D X 2
Introduction
T he u tility o f im m u n o c yto c h e m istry (IC C ) in diagnostic cytopa-
th o lo g y has added a new d im e nsion to cytologic diagnosis. The
ap p lication o f im m u n o h isto c h e m ic a l procedures to the w o rku p
o f d iffic u lt cases has paralleled the progress seen in surgical
pathology. IC C continues to play an im p o rta n t ro le in diag-
nostic cytop atholog y b u t is also increasingly im p o rta n t as an
ad juvant to o l in targeted therapies. W ith the use o f a uto m a tion ,
there is a great deal o f q u a lity im p ro ve m e n t o f ICC.
Immunocytology Techniques
The Specimen
Specim en Collection
The gam ut o f specimens th a t can be used fo r IC C includes exfo-
lia tive cell preparations, effusions, direct im p rints, fine-needle
aspirates, and th in -la yer collection samples. These specimens can
be processed th rou g h a ir d rying o r im m ed iate fix a tio n in alcohol
o r can be processed as cytocentrifuge o r cell b loc k preparations.
C yto m o rp h o lo g y fo rm s the basis fo r deciding the m ost appro-
priate anc illa ry technique to arrive at an accurate diagnosis. The
m ost im p o rta n t task o f the cytop atholog ist is to ensure th a t the
conventional R om anow sky o r Papanicolaou stains have been
exam ined, th a t a d iffe re n tia l diagnosis is generated, and th a t the
appropriate q uestion to be answered by IC C is form ulated .
Fixation
Prerequisites fo r adequate IC C studies1
are a well-spread film
o f cells o n a glass slide,2 adequate fix a tio n ,3 rem oval o f b lo o d
and
proteinaceous
m aterial,
and
a sensitive,
reproducib le
m eth od o f IC C .4 W e t fix a tio n in to alcohol m ust be perform ed
w ith o u t delay, because p artial a ir drying m ay result in false-
p ositive results. A ir-d rie d smears are often m ore cellular than
alcohol-fixed slides because som e m aterial o fte n floats o ff the
slide w h en it is alcohol-fixed. A ir-d rie d slides m in im iz e cell
loss and result in a m ore even film o f cells. Such slides m ust
be fixed im m ed iately before perform ance o f ICC, and the types
o f preferred fixatives vary am ong cytopathologists. C old acetone
and 95% alcohol are c om m o n fixatives,5 and B5 m ay be used
fo r ly m p h o id m arkers and neuroend ocrine antib od ies.6 Leong
and colleagues advocate the use o f in itia l fix a tio n in 0.1%
fo rm o l-s a lin e s o lu tio n , fo llo w e d b y fix a tio n in 95% ethanol
1043
previous page 1027 ComprehensiveCytopathology 1104p 2008 read online next page 1029 ComprehensiveCytopathology 1104p 2008 read online Home Toggle text on/off