Cell blocks in cytol (iac wrkshp i) 5 11-10
-
Upload
vshidham -
Category
Health & Medicine
-
view
5.194 -
download
0
description
Transcript of Cell blocks in cytol (iac wrkshp i) 5 11-10
![Page 1: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/1.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
May 17 & 19, 2010Vinod B. Shidham, MD, FRCPath, FIACProfessorCo-editor-in-chief & Executive editor, CytoJournal (www.cytojournal.com)Vice-chair Director of Cytopathology, Cytotechnology School, Cytopathology fellowship, & GI fellowshipDept of Pathology, Wayne State University Medical SchoolDetroit, MI 48201, [email protected]
Cell BlocksIn Cytopathology
(Workshop# 5 & 43)
![Page 2: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/2.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Disclaimer
VS is co-editor of ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company) cited for various methods of cell block preparation (the sketches and tables used are from this reference).
![Page 3: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/3.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
DisclaimerVS has indirect financial interest (through spouse) in AV marker mentioned in the workshop.
http://www.bioinnovationllc.com/Page_2.html
![Page 4: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/4.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Cell block- microbiopsy- Role in cytopathologic evaluation/patient managementCritical issues to be consideredDifferent methods of cell block preparationAligning the cells along the cutting surfaceDepth of section cuttingImmunophenotyping and cell blocks-Immunoreactivity interferenceMarker for SCIP approachA few study casesQ/A session
Outline
![Page 5: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/5.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Cell block of cytopathology specimen isEquivalent to microbiopsy evaluation
the interface between cytology and histopathology (bridge to histopathology/surgical pathology)
Routine example is Endocervical curretage (ECC), But without cytology preparation
Cell block with cytology preparation hasAdded benefit of excellent cytomorphologic details
in concert with architectural insight
Cyto-histo-pathology
![Page 6: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/6.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Role of cell blocks in cytopathologic evaluation/patient management
1. Immunophenotyping2. Special stains- Mucicarmine, Congo red, organism stains3. Architectural evaluation-
Trabecular-sinusoidal pattern in HCC,Hollow or solid proliferation spheres without cores
in carcinoma versus mesothelioma in effusions Evaluate for invasion Comparative evaluation with surgical pathology material
E.g- Peritoneal/pelvic washing Quantification of some features such as mitotic figures4. Enhanced sampling of FNAB rinses5. Molecular test e.g. FISH, CISH, In-situ PCR6. Archival for future studies
![Page 7: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/7.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Critical issues to be considered
Depending on the primary indication,the method of cell block preparation vary
Multiple variants should be considered for selecting the method and modifying it as needed for
individual specimens
Specimen type- Fresh versus fixed cells Cellularity of the specimenNature of cell distribution- predominantly solitary cells versus microfragments/aggregatesAncillary tests anticipatedAvailable resources/infrastructure in the labInstitutional and regional biases
![Page 8: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/8.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Different methods of cell block preparation
A. Cell block from specimen with clot or significant sediment- FNAB
B. HistoGel methodC. Gelatin embeddingD. Agar embedding methodE. Plasma-thrombin methodF. Collodion (Celloidin) bag method
G. From scraped material from cytology smearsH. From Millipore filtersI. From cells lifted selectively from the cytology preparation
(Kaneko C et al. Diagn Cytopathol 2000;22:117–119)
![Page 9: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/9.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Shidham & EppleCh 14 ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 10: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/10.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Processing of FNA aspirate to be submitted to laboratory for Cell block
Let the remaining aspirate clot in the syringe for 5 to 7 minutes (slightly longer than the clotting time).
1
Transfer the aspirated formalin with dislodged cot in to the specimen container with 10% formalin fixative
4
Gently and firmly remove the plunger of the syringe .
3
Aspirate 10% formalin from the container in which the specimen is to be submitted for cell block processing. This dislodges the clot from syringe wall.
2
![Page 11: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/11.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Protocol for plasma-thrombin method for cell block preparation
Shidham & EppleCh 14 ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 12: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/12.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Protocol for cell block preparation with collodion bag
Shidham & EppleCh 14 ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 13: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/13.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Protocol for preparing cell blocks with HistoGel
Shidham & EppleCh 14 ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 14: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/14.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Aligning the cells along the cutting surface
Depth of section cutting
![Page 15: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/15.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Varsegi GM, Shidham V (2009)Cell Block Preparation from Cytology Specimen with Predominance of Individually Scattered Cells. Journal of Visualized Experiments (JoVE) 2009 Jul 21;(29). pii: 1316. doi: 10.3791/1316. PMID: 19623160
![Page 17: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/17.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Video of JoVE article (8 minutes 15 sec)
At this stage, please note the URL
and observe the full FREE video in the new internet window
Video of JoVE article (8 minutes 15 sec)Video article FREE on web as open access at-
http://www.jove.com/index/Details.stp?ID=1316Or at
http://alturl.com/zkns
![Page 18: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/18.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
From:Varsegi GM, Shidham V (2009)Journal of Visualized Experiments (JoVE) 2009 Jul 21;(29). pii: 1316. doi: 10.3791/1316. PMID: 19623160
![Page 19: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/19.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
From:Varsegi GM, Shidham V (2009)Journal of Visualized Experiments (JoVE) 2009 Jul 21;(29). pii: 1316. doi: 10.3791/1316. PMID: 19623160
![Page 20: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/20.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Modified from:Varsegi GM, Shidham V (2009)Journal of Visualized Experiments (JoVE) 2009 Jul 21;(29). pii: 1316. doi: 10.3791/1316. PMID: 19623160
![Page 21: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/21.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
From:Varsegi GM, Shidham V (2009)Journal of Visualized Experiments (JoVE) 2009 Jul 21;(29). pii: 1316. doi: 10.3791/1316. PMID: 19623160
![Page 22: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/22.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Immunophenotyping and cell blocks-Factors affecting immunoreactivity- Loss, reduction, or enhancement of antigen immunoreactivityExposure to different reagents and fixative(s)TemperatureStorage of specimen with or without fixative
Subtractive Coordinate Immunoreactivity Pattern (SCIP) approachShidham & AtkinsonCh 5. Immunocytochemistry of effusion fl uids: introduction to SCIP approach. ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 23: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/23.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
23
1
65
4
8
7
Mes
oth
elia
l &
infl
amm
ato
ry c
ells
23
1
65
4
8
7
23
1
65
4
8
7
23
1
65
4
8
7
23
1
65
4
8
7
XM
etas
tasi
s(n
on
-car
cin
om
a)
2
3
1
6
54
7
2
3
1
6
54
7
2
3
1
6
54
7
3
1
6
54
7
2
3
1
6
54
7
ZM
etas
tasi
s(c
arci
no
ma)
21
54
3
7
6
21
54
3
7
6
21
54
3
7
6
21
54
3
7
6
21
54
3
7
6
Y
vim
en
tin
Pa
n C
K(M
ixtu
re o
f A
E1
/AE
3
& C
AM
5.2
)
Ca
lre
tin
in
WT
-1
LC
A(C
D45
)[o
r P
GM
1(C
D6
8)
or
mix
ture
of
LCA
&
PG
M1
]
A B C D E
SCIP approach
Immunocytochemistry of effusion fluids
(continued)
![Page 24: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/24.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Shidham & AtkinsonCh 5. ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 25: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/25.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Shidham & AtkinsonCh 5. ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
![Page 26: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/26.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
SCIP approach
F. CDX2Immunoreactivenuclear
HEstained cell blocksection
40X
B. Pan-cytokeratinImmunoreactive
C. LCA (CD45)Non-immunoreactive
A. VimentinNon-immunoreactive
D. CalretininNon-immunoreactive(Inset {2}- Mesothelial cell immunoreactivenuclear-cytoplasmic)
E. WT-1Non-immunoreactive(Arrow 2 with inset: Mesothelial cell- immunoreactivenuclear-cytoplasmic)
RM
RM
NC
10X
10X
10X
10X
10X
10X
40X
40X
40X
40X
40X
100X40X
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
NC
RM
RM
NC
Metastatic colonic adenocarcinoma, (peritoneal fluid).
35
![Page 28: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/28.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
A. Pap smear dx – LSIL, B. H&E cell block sections, C. p16 stained cell block sections, D. biopsy showing CIN II-III
A DCB
![Page 29: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/29.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
A. Pap smear dx – HSIL , B. H&E cell block section containing “microbiopsies”, C. p16 stained cell block section showing true nuclear positivity, D. biopsy showing invasive squamous cell carcinoma .
A DCB
![Page 30: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/30.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
12
a b CalretininCalretinin
Calretinin immunoreactivity pattern (epithelioid mesothelioma, pleural fluid). Mesothelioma cells (arrow in a) show nuclear (arrowhead 1) immunoreactivity usually with cytoplasmic immunostaining (arrowhead 2) imparting the so called ‘fried-egg’ appearance.
14
![Page 31: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/31.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
a b
c dCongo red stained 10 micron thick sections: Orange yellow birefringence under polarized light. The color changes to apple green when the axis of polarizer (blue arrows) is changed by 90 degree
Cell block- Fat pad aspiration
Positive control
![Page 32: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/32.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
C. CalretininNon-immunoreactive(Rare mesothelial cell [blue arrow] is immunoreactivenuclear-cytoplasmic)
D. BerEP4Immunoreactive
E. Estrogen receptorsImmunoreactive
B. CD68 (PGM1)Non-immunoreactive(inflammatory cells are immunoreactive)
A. VimentinNon-immunoreactive(Mesothelial & inflammatory cells are immunoreactive)
‘Subtractive coordinate immunoreactivity pattern’ (SCIP) in cell block sections
20X
20X
20X
20X
20X
40X
40X
40X
40X
40X
NC
RM
NC
NC
Metastatic mammary adenocarcinoma, (pleural effusion).
SCIP approach
(continued)
38
![Page 33: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/33.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
H&E CD177 vimentin
SMAWS cytokeratin S-100 protein
GIST
![Page 34: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/34.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
From: Çoban S et al. BMC Cancer 2004, 4:89Open access article from: http://www.biomedcentral.com/1471-2407/4/89
Cell block section of FNA of Hepatocellular Carcinoma
![Page 36: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/36.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Peer-reviewed, open access,
teaching material with many pictures.Hard copy and online availability.Opportunity for frequent updates
![Page 37: Cell blocks in cytol (iac wrkshp i) 5 11-10](https://reader033.fdocuments.us/reader033/viewer/2022052619/5564340dd8b42adb258b4b1c/html5/thumbnails/37.jpg)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Milwaukee Art Museum
Detroit
After June 1, [email protected]