cell blocks in cytol (iac wrkshp i) 5 11-10
DESCRIPTION
Role of cell blocks in cytopathologyTRANSCRIPT
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)
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).
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
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
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
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
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
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)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Shidham & EppleCh 14 ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
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
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)
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)
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)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Aligning the cells along the cutting surface
Depth of section cutting
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
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
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
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
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
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
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)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
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A B C D E
SCIP approach
Immunocytochemistry of effusion fluids
(continued)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Shidham & AtkinsonCh 5. ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Shidham & AtkinsonCh 5. ‘Cytopathologic Diagnosis of Serous Fluids’Elsevier (W. B. Saunders Company)
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
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
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
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
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
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
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
H&E CD177 vimentin
SMAWS cytokeratin S-100 protein
GIST
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
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
Vinod B. Shidham, MD, FRCPath, [email protected]
Cell BlocksIn
Cytopathology
Milwaukee Art Museum
Detroit
After June 1, [email protected]