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Purpose: The purpose of this study was to examine the diagnostic accuracy of the cell block (CB) method and clinical features affecting it in patients with vitreoretinal lymphoma (VRL). Methods: This study enrolled 38 eyes in 33 VRL patients, and 7 eyes in 7 patients with idiopathic uveitis who underwent diagnostic vitrectomy. Medical records including the results of CB cytology, interleukin (IL)-10/-6 concentrations, and immunoglobulin heavy chain gene (IgH) rearrangement were retrospectively searched. Results: Patients with VRL comprised 16 women and 17 men, and the age of onset ranged from 44 to 85 years (mean: 70 years). CB preparations detected large malignant cells in 35 eyes (92%), whereas the other 3 VRL eyes were negative. Two of the latter three eyes showed subretinal infiltrates, which existed in 7 of 35 CB-positive eyes. Intravitreal IL-10 and -6 concentrations were 1866 ± 4088 pg/mL and 98 ± 139 pg/mL, respectively, and the rate of IL-10/-6 >1 was 86.9%. The presence of IgH monoclonality was 63.2%. In patients with uveitis, CB specimens revealed no atypical but small inflammatory cells. IL-6 concentration was 311.1 ± 240 pg/mL, whereas IL-10 was undetectable in six eyes, and the IL-negative rate was 85.7%. Six eyes (85.7%) with uveitis showed no IgH monoclonality. Conclusions: Diagnostic accuracy of CB preparations in VRL could achieve an equivalent outcome to IL ratio calculation and IgH monoclonality detection. The appearance of subretinal infiltrates may diminish the CB positivity.

Background: Distinction between reactive mesothelial cells, malignant mesothelioma and carcinoma is challenging in both biopsy and cytologic material. This study was conducted to differentiate benign/ reactive mesothelial proliferation from malignant mesothelial proliferations and metastatic adenocarcinoma by using immunohistochemical (IHC) markers Desmin (DES), Epithelial membrane antigen (EMA) and Calretinin (CAL) in pleural uid cell block (CB) preparations. A two year descriptive study (Oct.2016- Sept.2018). 46 pleural uids samples sentMethods : to the Dept. of Pathology, RIMS for routine examination and histopathological examination by CB preparation were studied using IHC markers EMA,DES and CAL following H & E stain. Out of 46 cases, 9(19.6%) cases were diagnosed as benign, 23(50.0%) as reactive andResults: 14(30.4%) as adenocarcinoma on H & E section by CB preparations within an age range 34 to 80 years (Mean±SE, 60.32±12.13). Following IHC staining with EMA, DES & CAL, 11(23.9%) cases were conrmed as benign, 17(37.0%) as reactive, 16(34.8%) as adenocarcinoma and 2(4.3%) cases as malignant mesothelioma. This study showed that EMA, DES and CAL helpful in conrming benign or reactive mesothelialConclusions: and malignant mesothelial with epithelial cells which will be helpful in providing appropriate diagnosis in difcult cases and provide better patient management.

BACKGROUND Inconclusive cytomorphology often results due to failure to distinguish between adenocarcinoma cells from benign, reactive, atypical mesothelial cells in effusion specimens. To resolve such dilemmas, auxiliary techniques like immunohistochemistry were utilised to reach a definitive diagnosis for better treatment and management of patients. We wanted to compare cytodiagnosis achieved on cell block preparations with the cytodiagnosis on conventional smear and perform immunohistochemistry for epithelial membrane antigen (EMA), calretinin, desmin, vimentin and E-cadherin on cell block preparation of the fluids in cases of indistinguishable cytomorphology of adenocarcinoma and reactive, atypical, and benign mesothelial hyperplasia. METHODS The immunohistochemical markers namely EMA, calretinin, vimentin, desmin and Ecadherin were applied on cell blocks employing streptavidin-biotin method. Immunohistochemistry was interpreted by giving scores to the percentage of stained cells. RESULTS EMA and E-cadherin had 100 % sensitivity in diagnosing adenocarcinoma whereas calretinin, vimentin and desmin were 100 % sensitive in diagnosing reactive, atypical mesothelial carcinoma on the cell block preparations. CONCLUSIONS Immunocytochemistry of fluid should be carried out on the cell block preparation where cytological diagnosis on conventional smear and cell block fails to detect malignant cells in the preparation. KEY WORDS Cell Block, Adenocarcinoma, Mesothelial Cells, Immunohistochemistry, EMA, Calretinin, Vimentin, Desmin, E-Cadherin

Background: Programmed death ligand-1 (PD-L1) assessed by immunohistochemistry (IHC) is used as biomarker for pembrolizumab therapy in advanced stage lung cancer patients. However, data permitting direct performance comparison between cytology and surgical specimen types are limited since both specimens from a single tumor site are infrequently available. In addition, alcohol fixation used with cytology specimens requires technical validation of the PD-L1 IHC assay before clinical use. We here report our experience with implementation of the PD-L1 22C3 IHC pharmDxTM assay for cytologic samples at a large tertiary cancer center. Study Design: Archival formalin-fixed (FF), paraffin-embedded cell blocks (CBs) and subsequent lung tumor resections (LTRs) from the same anatomical site were used for a direct comparison of PD-L1 tumor proportion scores (TPSs). TPS values were independently determined by one surgical lung pathologist and two cytopathologists blinded to the specimen pairs. An interim analysis was performed to facilitate the pooling of expertise among observers. After PD-L1 22C3 IHC pharmDxTM implementation for FF cytology specimens, dual-processed samples were used for a prospective technical validation of CytoLyt® prefixation (CF). Digital image analysis was performed for a subset of dual-processed specimens. Results: Eighty-one CBs and LTRs were included for comparison of the specimen types. PD-L1 assessment in CBs had an accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of 88.9/72.8, 66.7/73.5, 95.2/72.3, 80.0/65.8, and 90.9/79.1% for the ≥50/≥1% cutoff, respectively. The intraclass correlation coefficient was 0.84 (95% confidence interval [CI]: 0.76, 0.90), and it improved after interim analysis (before: 0.79 and after: 0.92). The overall concordance between CF and FF for the categories defined by the ≥50/≥1% cutoff values was 90.4% (95% CI: 79.0, 96.8). Similar assay performance was confirmed by digital analysis. Conclusions: PD-L1 22C3 IHC pharmDxTM shows good reliability if used with CB preparations. CF does not impact assay results significantly. Clinical validation with outcome data is needed, and digital methods of assessment should be further investigated.

Objective Primary retroperitoneal involvement of lymphoproliferative disorders is rare. Diffuse large B cell lymphoma (DLBCL) in the retroperitoneum has been scarcely reported and the diagnostic workup is challenging due to difficulties in access for tissue acquisition. Materials and methods A 73-year-old female presented with epigastric pain and nausea. A computed tomography scan revealed retroperitoneal lymphadenopathy. An EUS-FNA was performed and a cytological diagnosis of lymphomatous disorder was made on direct smears .A panel of immunocytochemical markers was applied on cell block preparations and revealed a EBV related DLBCL (CD20 positive, PAX-5 positive, CD3 negative, MUM-1 positive, BCL-2 positive, BCL-6 positive, CD10 positive, MIB-1 positive, and LMP-1 positive). Conclusion Our case illustrates the utility of EUS-FNA in the work-up of retroperitoneal lymphadenopathy and the importance of cell block processing in the diagnostic approach of EBV related DLBCL.

Background The identification of BRAF mutations in thyroid cancer has prognostic and therapeutic implications. Although the gold standard for identifying BRAF mutations is molecular testing, the ability to perform BRAF p.V600E immunostaining on fine-needle aspiration (FNA) samples can facilitate the rapid triaging of patients to treatment options. Methods A total of 50 thyroid carcinoma FNA samples, including papillary (29 samples), poorly differentiated (10 samples), anaplastic (9 samples), and Hurthle cell (2 samples) carcinomas, with a known BRAF p.V600E mutation status were selected for the current study. Immunostaining was performed on smears and cell block sections using an anti-BRAF p.V600E antibody (clone VE1). The results were compared with the known mutation status obtained by molecular testing and/or immunostaining of surgical pathology material from the same patient. Results Of the total of 50 cases, 26 cases had smears available for the evaluation of BRAF p.V600E immunostaining; positive immunostaining was noted in 16 samples and negative immunostaining was noted in 4 samples, whereas 6 cases were equivocal. Of the 34 cases for which cell blocks were available for evaluation, BRAF p.V600E immunostaining was positive in 17 cases, negative in 16 cases, and equivocal in 1 case. The overall sensitivity and specificity of BRAF p.V600E immunostaining on the cell block preparation was 94.4% and 100%, respectively, whereas for the smears it was 80% and 63.6%, respectively. Conclusions BRAF p.V600E immunostaining can be performed reliably on thyroid FNA cell block preparations. However, false-positive results on direct smears limit their utility and therefore need to be interpreted with caution. Cancer Cytopathol 2018;126:406-13. © 2018 American Cancer Society.

BACKGROUND: Programmed death ligand-1 (PD-L1) assessed by immunohistochemistry (IHC) is used as biomarker for pembrolizumab therapy in advanced stage lung cancer patients. However, data permitting direct performance comparison between cytology and surgical specimen types are limited since both specimens from a single tumor site are infrequently available. In addition, alcohol fixation used with cytology specimens requires technical validation of the PD-L1 IHC assay before clinical use. We here report our experience with implementation of the PD-L1 22C3 IHC pharmDxTM assay for cytologic samples at a large tertiary cancer center. STUDY DESIGN: Archival formalin-fixed (FF), paraffin-embedded cell blocks (CBs) and subsequent lung tumor resections (LTRs) from the same anatomical site were used for a direct comparison of PD-L1 tumor proportion scores (TPSs). TPS values were independently determined by one surgical lung pathologist and two cytopathologists blinded to the specimen pairs. An interim analysis was performed to facilitate the pooling of expertise among observers. After PD-L1 22C3 IHC pharmDxTM implementation for FF cytology specimens, dual-processed samples were used for a prospective technical validation of CytoLyt® prefixation (CF). Digital image analysis was performed for a subset of dual-processed specimens. RESULTS: Eighty-one CBs and LTRs were included for comparison of the specimen types. PD-L1 assessment in CBs had an accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of 88.9/72.8, 66.7/73.5, 95.2/72.3, 80.0/65.8, and 90.9/79.1% for the ≥50/≥1% cutoff, respectively. The intraclass correlation coefficient was 0.84 (95% confidence interval [CI]: 0.76, 0.90), and it improved after interim analysis (before: 0.79 and after: 0.92). The overall concordance between CF and FF for the categories defined by the ≥50/≥1% cutoff values was 90.4% (95% CI: 79.0, 96.8). Similar assay performance was confirmed by digital analysis. CONCLUSIONS: PD-L1 22C3 IHC pharmDxTM shows good reliability if used with CB preparations. CF does not impact assay results significantly. Clinical validation with outcome data is needed, and digital methods of assessment should be further investigated.

Introduction The diagnosis of pancreatic ductal adenocarcinoma (PDA) on endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) material is often challenging. An immunohistochemical (IHC) panel may help establish the diagnosis of PDA in cases limited by sample size or ambiguous cytology. S100P, IMP3, and SMAD4 are 3 IHC markers that have shown promise as individual markers for PDA that have never been tested together as a panel. In this study, we evaluated the individual and combined efficacy of S100P, IMP3, and SMAD4 for the detection of PDA. Materials and Methods S100P, IMP3, and SMAD4 IHC staining was performed on cell blocks (CBs) procured from pancreatic EUS-FNA procedures. The cohort included CBs that were diagnostic for PDA (n = 35), suspicious but nondiagnostic for PDA (n = 2), as well as CBs with benign pancreatic ductal epithelium (n = 12) and benign reactive pancreatic ductal epithelium (n = 18). A positive result for IMP3 and S100P was defined as moderate or strong staining of >10% of ductal cells. Complete lack of SMAD4 nuclear staining was considered a positive result—any nuclear SMAD4 staining was considered a negative result. Results Two and 3 IHC marker panels were almost always more specific than individual IHC markers. Positivity for at least 2 of 3 IHC markers was a sensitive (91.89%) and highly specific (100%) marker of PDA. Conclusions The 3 IHC marker panel composed of S100P, IMP3, and SMAD4 is highly specific for PDA. Future studies should evaluate efficacy in a cohort with more atypical and suspicious cases.

Studies using the cell-block technique for bone marrow (BM) analysis are limited in the veterinary literature. This work assessed whether the histopathological analysis of canine BM was feasible using cell-block cytoinclusions prepared from fine-needle sternal aspirate samples. Eight clinically healthy young-to-middle-aged dogs underwent both fine-needle sternal aspiration for BM cell-block (BM-Cb) processing and iliac-crest BM core biopsy (BM-B). Histopathologic parameters were compared between the 2 methods. There were no statistically significant histopathological differences between hematopoietic tissue areas (P = .6294) in the BM-Cb and BM-B sections, and they had similar microscopic characteristics and microarchitecture. Cellularity and reticulin-fiber staining were equivalent in the BM-Cb and BM-B preparations in 87.5% (7/8) and 100% (8/8) of the sections, respectively. However, the quantitative results of the megakaryocytic series differed between BM-Cb and BM-B in 37.5% (3/8) of the sections, and the myeloid:erythroid (M:E) ratios differed between the 2 methods in 25% (2/8). These preliminary data indicate that cell-block preparations made from sternal fine-needle aspiration samples warrant continued evaluation in a larger number of animals, including those with various diseases affecting the bone marrow.

Background: Cell Block (CB) procedures have now become an established part of cytological diagnostics because of its pivotal role in diagnosis and ancillary studies. Hence the present study was undertaken to emphasize the role of CB technique over Conventional Smear (CS) in serous effusions and to compare the Plasma-Thrombin (PT) block to Formalin Method block (FM) in assessment of morphological preservation and cellularity. Aim: To obtain simple , cost effective and ideal CB preparation where in maximal number of cells are displayed within a small area Methods: The sample was divided into three Parts(A,B,C). After centrifugation of all three parts of sample at 3000rpm for 15min – Part A sediment was used to prepare two CS for Papanicolaou (PAP) and May Grunwald Giemsa(MGG) stains. Part B sediment was subjected for 1hr and 24hr fixation in 1:1 solution of 5ml ethylalcohol and 10% formalin. To the Part C sediment 2drops of finger prick plain blood was added, mixed well and allowed to clot. The sediment of Part B and the clot of Part C were then processed for paraffin embedding. Result: 110 fresh effusion samples were evaluated for cellularity retention of architectural patterns and volume of background. FM block’s were inconclusive in 12 cases due to low cellularity. PT block’s were all evaluable with best preservation of architecture and pale background. Conclusion: The CB technique revealed better architectural patterns and increased the sensitivity of cytodiagnosis. PT block’s had sufficient to abundant cellularity with evenly distributed cells in small area. PT preparation is simple and cost effective. DOI: 10.21276/APALM.1432

Objective Atypical squamous cells of undetermined significance (ASCUS) and atypical glandular cells (AGC) reflect cellular abnormalities insufficient for clear diagnosis. We used cell pellets obtained from liquid-based cytology (LBC) to prepare cell blocks (CB) and clarify the initial diagnosis of ASCUS and AGC. Study Design A total of 393 CBs with initial diagnosis of ASCUS or AGC were processed. Of those, 305 of the ASCUS and 20 of the AGC had adequate specimens. We compared results of CBs prepared from ASCUS and AGC to determine which had higher frequencies of higher-grade lesions. Results A majority of specimens (83%) were adequate for evaluation. Compared with the initial diagnosis, 14% (42/305) of ASCUS were diagnosed with low-grade squamous intraepithelial lesion (LSIL) in CB, while 10% (2/20) of AGC were diagnosed with LSIL or adenocarcinoma. No statistical relationship between the initial diagnosis of ASCUS and AGC and results of higher-grade lesions in CB is evident as determined by p value greater than 0.05 (p = 0.228). Conclusions CBs prepared from Liqui-PREP cell pellets are, in most cases, assessable and can be useful as an adjunctive test to help clarify the initial diagnosis of ASCUS and AGC. Diagn. Cytopathol. 2017;45:520–525. © 2017 Wiley Periodicals, Inc.

Myofibroblastoma (MFB) is a benign tumor of the mammary stroma with predominant myofibroblastic differention. The cytologic reports of MFB are very few in the available literature. From the cytodiagnostic point of view about 21 cases of MFB with cytological evaluation by fine-needle aspiration cytology (FNAC) have been encountered in the English-language literature: A 35-year-old woman presented with lump in the left breast. FNAC showed mild degree of pleomorphism with occasional groups with fibrous stroma and tumoral cells. A few benign epithelial cell groups were seen. Hematoxylin-eosin-stained sections of cell block preparation from the aspirate showed fascicles of spindle cells forming whorl structures. Three months later, excision biopsy was performed. The diagnosis was a classic variant of MFB. On immunohistochemical examination, sections were found to be highly positive for vimentin, CD34, and bcl- 2. In light of these findings, cell block material was retrospectively reviewed both morphologically and immunhistochemically. The findings of resection and cell block material were found to be very similar. MFB may cause a potential diagnostic pitfall while interpreting FNAC due to its wide differential diagnosis spectrum. We concluded that cytology and cell block findings complement each other. Diagn. Cytopathol. 2016;44:1064-1069. © 2016 Wiley Periodicals, Inc.

Objectives Programmed death-ligand 1 (PD-L1) is an emerging molecular target in anticancer therapy, most notably non–small cell lung cancers. PD-L1 expression in pancreatic adenocarcinomas (PDAs) on surgical specimens is highly variable, with 10% to 60% of tumors showing expression. PD-L1 expression in PDA on endoscopic ultrasound-guided fine needle aspirations (EUS-FNAs) has been rarely studied. Methods Formalin-fixed, paraffin embedded (FFPE) cell blocks (CBs) from 65 EUS-FNA samples of 55 patients, with a diagnosis of PDA, with at least 20% tumor cellularity were retrieved. The cell blocks were originally fixed in CytoRich fixative. Immunohistochemical staining (IHC) for PD-L1 was performed using the M365329-1 (22C3) clone according to the manufacturer’s approved protocol and optimized for the fixation method using appropriate controls. A combined positive score (CPS) defined per CAP guidelines as the total number of positive tumor cells and inflammatory cells as a percentage of the total number of tumor cells was assessed for each case and was grouped as 20. Results Twenty-five samples (38%) from 21 patients showed immunoreactivity to PD-L1, with 21 (32%) having a CPS of Conclusion Immunohistochemical analysis for PD-L1 is feasible on EUS-FNA CB samples when optimized and validated for the fixation method using appropriate controls. PD-L1 expression is seen in only a minority of PDAs, albeit with a very low CPS. The potential role of PD-L1 as a prognostic marker for disease progression needs further exploration.

Objectives Immunohistochemistry especially for biomarkers such as PD-L1 in personalized medicine is increasingly being performed on cell blocks from cytopathology samples. We evaluated the effects of staining cell blocks (CBs) from different cytologic fixatives using four different commercially available PD-L1 antibody clones. Methods PD-L1 immunohistochemistry (IHC) using four different commercially available clones was performed on eight cell blocks processed from two different fixatives and also on eight formalin-fixed tissues. The clones used were 22C3 at 1:50 dilution and 1:100 dilution, SP263, SP142, and E1L3N. The cell block (CB) samples contained cells that strongly express PD-L1 and either fixed in CytoLyt (methanol-based fixative, n = 4) or CytoRich (ethanol-formalin-based fixative, n = 4). Formalin-fixed control tissues (tonsillar tissue, n = 4; stomach, n = 4) were also included. Results Expression for PD-L1 was noted on all the CytoRich fixed cell blocks (n = 4) for all four antibody clones, with the intensity and distribution of expression comparable to the formalin fixed tissues (n = 8). However, consistent absence of expression for PD-L1 was noted on all the CytoLyt fixed cell blocks (n = 4) for all four antibody clones. Conclusion The results of this pilot study demonstrate that PD-L1 IHC on cell blocks is feasible. However, there is a need to validate IHC protocols according to specific fixation methods.

Objectives: HER2 status of breast carcinoma is a powerful prognostic and predictive biomarker-particularly in the metastatic setting. Limited data is available regarding assessment of HER2 on cell block preparations (CBP). The primary objective of this study was to assess correlation between HER2 results obtained via immunohistochemistry (IHC) and fluorescence in-situ hybridization (FISH) in cases of metastatic breast carcinoma (MBC) on CBP. Secondary objectives included study of inter-observer variability in interpretation of HER2 on IHC, and concordance between HER2 results on CBP and formalin-fixed paraffin embedded material (FFPEM). Materials and Methods: Cases of MBC diagnosed on fine needle aspirates (FNA) with HER2 testing performed via IHC and FISH on CBP over 5 years (2010-2015) were reviewed. CBP material was fixed in an ethanol-based fixative (CytoRich Red Fixative system, BD). HER2 IHC was performed using polyclonal antibodies against Cerb-2 (Dako 0485). HER2 FISH testing was performed using the LSI HER2/neu/CEP17 probes (Vysis/Abbott Molecular Inc., Des Plaines, IL). Results: Seventeen cases (all female, median age: 59) were analyzed. 41% of CBP were products of bone FNA (7/17). Other sites included lymph node (3), lung (2), pleural fluid (2), liver (1), skeletal muscle (1) and mesentery (1). Median interval between diagnosis of primary carcinoma and FNA of metastasis was 5 years (range: 10 months-32 years). FISH was inconclusive due to suboptimal specimen quality in 2 cases. Correlation between IHC and FISH results was as follows: IHC 0/1+ (0/2; 0% amplification), IHC 2+ (2/12; 16.7% amplification) and IHC 3+ (0/1; 0% amplification). Inter-observer agreement of IHC scoring between 2 pathologists who independently reviewed IHC slides was fair (66.7% agreement, κ = 0.31). Comparison of HER2 results on CBP with FFPEM (primary carcinoma or metastasis) showed a high discordance and slight agreement (discordance rate = 37.5%; κ = 0.02). Conclusions: In this study, (a) 16.7% of MBC cases that scored 2+ on IHC showed amplification on FISH, (b) there was poor inter-observer agreement in HER2 scoring of IHC on CBP, and (c) there was high discordance between HER2 results obtained on CBP and FFPEM. Our results indicate that HER2 testing of MBC on CBP may be unreliable.

The effectiveness of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) has been established for a definite pathological diagnosis of pancreatic solid masses. We investigated the usefulness of cell block preparations in EUS-FNA, by evaluating the added value of cell block preparations over conventional smears alone.Between March 2011 and June 2013, 61 patients were retrospectively evaluated who underwent EUS-FNA for pancreatic solid masses. Diagnostic values for diagnosing pancreatic malignancy were compared for a combination of the conventional smear and cell block (CSCB) and the conventional smear alone (CS).The addition of the cell block technique increased the sensitivity of conventional smear for diagnosing the pancreatic malignancy from 79% (CS) to 90% (CSCB, p=0.0313) and the accuracy from 81% to 91% (p=0.0313). The specificity and positive predictive value were 100% in both methods. The negative predictive value was increased from 33% (CS) to 50% (CSCB), but this difference was not statistically significant (p=0.0833).The addition of the cell block method after a conventional smear may increase the sensitivity and negative predictive value for diagnosing the pancreatic malignancy in patients with pancreatic masses who undergo EUS-FNA. Further study may be warranted to determine whether the cell block method can replace the conventional smear.

Atypical squamous cells of undetermined significance (ASCUS) and atypical glandular cells (AGC) reflect cellular abnormalities insufficient for clear diagnosis. We used cell pellets obtained from liquid-based cytology (LBC) to prepare cell blocks (CB) and clarify the initial diagnosis of ASCUS and AGC.A total of 393 CBs with initial diagnosis of ASCUS or AGC were processed. Of those, 305 of the ASCUS and 20 of the AGC had adequate specimens. We compared results of CBs prepared from ASCUS and AGC to determine which had higher frequencies of higher-grade lesions.A majority of specimens (83%) were adequate for evaluation. Compared with the initial diagnosis, 14% (42/305) of ASCUS were diagnosed with low-grade squamous intraepithelial lesion (LSIL) in CB, while 10% (2/20) of AGC were diagnosed with LSIL or adenocarcinoma. No statistical relationship between the initial diagnosis of ASCUS and AGC and results of higher-grade lesions in CB is evident as determined by p value greater than 0.05 (p = 0.228).CBs prepared from Liqui-PREP cell pellets are, in most cases, assessable and can be useful as an adjunctive test to help clarify the initial diagnosis of ASCUS and AGC. Diagn. Cytopathol. 2017;45:520-525. © 2017 Wiley Periodicals, Inc.

Evaluation of ER, PR and Her 2 are routinely performed on breast carcinomas. For accurate detection of these markers, compliance with the ASCO/CAP guidelines is recommended. Our previous study showed that alcohol fixation did not affect ER results when alcohol-fixed cell block (CB) sections were compared to formalin-fixed tissue sections, while PR and Her2 showed less concordance. The aim of this study was to evaluate and to compare ER, PR and Her2 IHC results on formalin-fixed CB sections to those observed on subsequent surgical (needle core or resection) specimens (SS). Fifty cases of formalin fixed CB samples obtained from primary (18%) and metastatic (82%) breast carcinomas were studied, all of which had subsequent SS available. ER, PR, and Her2 IHC studies were done on all samples and results were compared. ER results on formalin-fixed CB samples showed excellent correlation with SS (correlation coefficient cc = 0.82). While there was minimal improvement in PR results (cc = 0.433), Her2 detection did not improve by formalin fixation (cc = 0.439). Formalin fixation for CB preparations does not significantly improve the already good detection of ER positive breast tumors. The concordance rate in PR and IHC results between formalin-fixed CB and SS samples showed improvement as compared with the alcohol-fixed CB results. However, there was no improvement in detection of Her2 overexpression by using formalin fixation on cytology specimens.

BACKGROUND Cell block (CB) preparation during the endobronchial ultrasound-guided transbronchial fine-needle aspiration (EBUS-TBNA) procedure plays an important role in the diagnosis of lung cancer and recovery of cellular material for molecular characterization of the tumor. However, the efficiency of the conventional method of CB preparation is suboptimal.

Objective Recent studies have validated the use of cytologic materials to determine HER-2/neu status. Good concordance has been shown between results obtained by immunocytochemistry (ICC), immunohistochemistry (IHC), fluorescence in situ hybridization (FISH) on cytologic and surgical specimens. However, the utility of cytologic cell block material in determining HER-2/neu status has not been reported and is the subject of this study. Study Design HER-2/neu status was determined in 25 cases of primary or metastatic breast carcinoma by IHC and FISH. All cases were formalin-fixed, paraffin-embedded (FPPE) cell block preparations. ICC was performed using monoclonal antibodies TAB250 (Zymed) and CB11 (Novacastra Laboratories). FISH analysis was performed using the PathVysion HER-2 probe kit (Vysis, Inc.). Results of ICC and FISH were compared in each case. Results Of 25 cases studied, 17 showed no protein overexpression or amplification. Five cases showed protein overexpression and amplification. The remaining 3 cases showed 2+ staining intensity by ICC in 10, 20, and 50% of carcinoma cells, respectively, and all demonstrated lack of amplification. Conclusion Immunocytochemistry performed on FFPE cell block material is a reliable method for determining HER-2/neu status in cytologic specimens. We recommend routine preparation of FFPE cell block material in instances of suspected primary of metastatic breast carcinoma.

BACKGROUND Calretinin (CR) is a valuable marker in the immunohistochemical distinction between malignant mesothelioma (MM) and adenocarcinoma (ACA) in tissue sections. However, there is limited and conflicting data regarding the utility of CR in this differential diagnosis on cytologic material, especially cell block preparations. Also, the possible role of CR in the distinction of papillary serous borderline tumor (SBT) cells from reactive mesothelial cells in peritoneal washings has not been examined. METHODS Formalin-fixed paraffin-embedded cell block specimens of cytologic fluids, washings, and aspirates with a suspicious or positive cytologic diagnosis and a confirmed diagnosis of MM (29 cases), ACA (39 cases), and SBT (10 cases) were used for CR immunohistochemistry (IHC). RESULTS Moderate to strong staining in > 50% of tumor cells was noted in 26 of 29 (90%) MMs but in only 3 of 39 (8%) ACAs and 1 of 10 (10%) SBTs. Admixed reactive mesothelial cells (when present) were strongly positive in all fluid specimens, but the staining pattern of benign non-reactive mesothelial cells in washing specimens was less reliable. CONCLUSIONS CR is both a sensitive and specific marker of reactive and neoplastic mesothelial cells in cytologic cell block preparations and is thus useful in the differential diagnosis of MM and metastatic ACA in malignant effusions. However, CR IHC does not appear to allow definitive identification of SBT in peritoneal washings. Cancer (Cancer Cytopathol) 2000;90:312–9. © 2000 American Cancer Society.

OBJECTIVE: To evaluate the utility of immunohistochemical stains for desmin in discriminating mesothelial cells from adenocarcinoma in serous fluid cell block preparations. STUDY DESIGN: Cell block preparations from 22 cases (representing 18 patients) that were positive for carcinoma and 5 cases that were negative for malignancy were immunostained with an antibody to desmin. Positive staining was evaluated and scored semiquantitatively in both tumor cells and background mesothelial cells in the malignant cases and mesothelial cells in the negative controls. Staining was evaluated with a score of 0-3 for intensity and 0-5 for distribution. The sum of the two scores was recorded as the total score (TS). RESULTS: Mesothelial cells from all the carcinoma and benign cases stained with desmin (median TS = 5.5, range 4-8), typically strong in intensity and widespread in distribution. Positivity was observed in carcinoma cells in all cases, typically weak and focal (range 2-4). Using a total score of 4 as a cutoff for definitively positive staining, desmin staining was positive in mesothelial cells in 25/25 cases and carcinoma cells in 1/22 cases (P

Objective: Cervical cancer is one of the most common malignancies worldwide, yet it is preventable by population screening. In a previous study, we confirmed the feasibility of utilizing whole slide imaging (WSI) of cell block (CB) preparations to overcome the limitations of digitizing cytologic samples. In this study, we evaluated the accuracy of WSI in identifying various organisms and nonneoplastic findings. Study Design: A total of 335 WS images from Pap CB preparations were analyzed using the Aperio system. The test performance characteristics of ThinPrep (TP) and WSI samples were compared for adequacy, for the presence of bacterial vaginosis (BV), fungi, Trichomonas vaginalis (TV) and herpes simplex virus (HSV) and for nonneoplastic findings. Results: The WSI samples contained optimal material from all preparations. BV was diagnosed in 33 WSI versus 36 TP samples. Budding yeasts and/or pseudohyphal forms were noted in 18 WSI versus 19 TP samples. TV organisms (10 of 11 samples) and 1 HSV case were accurately identified in the WSI and TP samples. Squamous metaplasia, keratosis and reactive/reparative and inflammatory changes were easily identified by WSI. Conclusions: The concept of WSI from Pap CB preparations is potentially feasible for adoption. Digital remote web-based technology eliminates the need for an individual on site, saving time and resources.

We compared the results of smears to those of cytospin and cell block preparations from fine-needle aspirations to determine the cost effectiveness of each and to determine which should be routinely obtained. We reviewed 844 cases, 361 of which had both smears and cytospins, and 483 of which had both smears and cell blocks. Smears alone were diagnostic in 94% of cases (796/844 cases), cytospins alone diagnostic in 43% of cases (154/361 cases), and cell blocks alone diagnostic in 57% of cases (277/483 cases). Cytospins contributed additional information beyond that obtained from smears in 2% (7/361) and cell blocks in 12% (57/483) of cases. When smears were nondiagnostic, cytospins contributed additional information in 10% (2/21) of cases and cell blocks contributed additional information in 44% (12/27) of cases. The cost of providing a diagnosis from smear alone is $212, from cytospin alone is $352, and from cell block alone is $392. The cost for additional information established by cytospin is $7,736 and by cell block the cost is $1,906. Smears are superior to either cytospins or cell blocks in providing a diagnosis. It is not cost-effective to obtain either cytospins or cell blocks in addition to smears on all cases. However, it is cost-effective to obtain cell blocks when the immediate smear evaluation is nondiagnostic.

Objective To evaluate the utility of CD10 immunostaining on cell block preparations from pelvic washing cytology material obtained during laparoscopy in cases of pelvic endometriosis (PE). Study design Six premenopausal women presenting with ovarian masses underwent a laparoscopic procedure. In addition to routine cytospin preparations, cell blocks were prepared using the formalin-fixed, paraffin-embedded method from the pelvic washings in each case. Immunocytochemistry using monoclonal antibodies for cytokeratin (CK) 7, CK20, CD10, estrogen receptor (ER), progesterone receptor (PR) and CA125 was performed on sections from cell blocks using the streptavidin-biotin peroxidase complex. Results Clusters of epithelial cells (3 cases) and stromal cells (2 cases) were seen in a background rich in hemosiderin-laden macrophages and poorly preserved mesothelial cells on cytospin smears. CD10 and CK7 were consistently positive in the stromal and epithelial components, respectively, in all cases on cell block sections. Variable staining reactions were noted for ER, PR and CA125. Conclusion CD10 immunostaining is a useful ancillary method in the diagnosis of endometriosis. Its routine use on cell block preparations from pelvic washings in women undergoing a laparoscopic procedure is recommended.

Endobronchial ultrasound-guided transbronchial needle aspiration has demonstrated its accuracy in the diagnostic workup of enlarged mediastinal lymph nodes. In addition to conventional smears, the use of liquid-based cytology (LBC) and cell block preparations (CBP) has been introduced more recently. The aim of our study was to determine the performance of each of the different techniques, separately and combined, in terms of diagnostic yield and sensitivity. A total of 290 consecutive patients were included. The pathological examination was based on smear cytology, LBC, and CBP. Adequate sampling was defined by the presence of pathological material or lymphocytes. The global diagnostic yield was 82.7 % and the sensitivity was 89.1 %. The diagnostic yield was 72.8 % for smears, 78.8 % for LBC, and 69.9 % for CBP. The combination of smears with CBP significantly increased diagnostic yield (p = 0.01) and sensitivity (p = 0.006), but not the combination of smears with LBC (yield: p = 0.07; sensitivity: p = 0.13). The combination of the three techniques further increased yield (p = 0.007) and sensitivity (p = 0.006), compared with smears alone. CBP were more sensitive than smears for both diagnoses of carcinoma (p = 0.01) and granulomatous inflammation (p = 0.048). Conversely, LBC was less sensitive than smears for granulomatous inflammation (p = 0.004), but the difference was not significant for carcinoma (p = 0.42). CBP, as a complement to smears, increases diagnostic yield and sensitivity for both diagnoses of carcinoma and granulomatous inflammation. LBC, if used alone, increases the risk of a false-negative result.

Cell block (CB) preparation during the endobronchial ultrasound-guided transbronchial fine-needle aspiration (EBUS-TBNA) procedure plays an important role in the diagnosis of lung cancer and recovery of cellular material for molecular characterization of the tumor. However, the efficiency of the conventional method of CB preparation is suboptimal.In the current study, the "tissue coagulum clot" cell block (TCC-CB) method was used to prepare the CBs and its efficiency was compared with that of the conventional saline rinse cell block (NR-CB) method. A total of 84 consecutive TCC-CBs (106 lymph nodes [LNs] and 14 lung lesions) and 28 consecutive cases of NR-CB (39 LNs and 3 lung lesions) obtained within the same time period were included in the current study.In the TCC-CB specimens, 94 of 106 LN cases (88.7%) yielded sufficient diagnostic material, as did 11 of 14 lung lesions (78.6%). In the NR-CB group, which was used as the control, 22 of 39 LN specimens (56.4%) and none of 3 lung specimens (0%) were found to provide sufficient diagnostic material. Although the average size of the LNs in the study group were not significantly different from those in the control group (1.76 cm vs 1.82 cm; P.05), the overall nondiagnostic rates in the TCC-CB and NR-CB groups were 11.2% and 43.6%, respectively (P.001). The nondiagnostic rates of the lung specimens were 15.4% in the TCC-CB group and 100% in the NR-CB group (P.05). In addition, immunohistochemistry studies and epidermal growth factor receptor (EGFR)/KRAS mutational analyses were performed in 26 and 14 TCC-CB cases, respectively. With the exception of 1 case, all of them had satisfactory results.The data from the current study demonstrate that the TCC-CB method significantly increases the cellular yield of CB preparations without compromising cytomorphological characterization of tumor cells.

Objective: To determine the value of calretinin and cytokeratin (CK) 5/6 in discriminating mesothelioma from adenocarcinoma in serous effusion specimens. Methods: A total of 101 recent, histologically or clinically confirmed malignant effusions with immunostained cell block preparations were reviewed. The cases consisted of 34 mesotheliomas and 67 adenocarcinomas. This included 17 ascitic fluid and 84 pleural fluid samples. The adenocarcinomas included metastatic carcinomas from the breast (12), lung (19), stomach (3), colon (1), pancreas (2), ovary (6) endometrium (1) and 23 histologically confirmed metastases from unknown primary sites. The cases were assessed as negative or positive (>5% of cells stained). The staining pattern was recorded as cytoplasmic, cell membrane, nuclear or cytoplasmic and nuclear staining. Results: Calretinin staining was present in 97% (33/34) of the mesothelioma cases with a majority of them showing both cytoplasmic and nuclear staining (29/33). Only 3% (2/67) of adenocarcinomas were positive for calretinin, one being a lung adenocarcinoma and the other an adenocarcinoma of unknown primary site in an ascitic fluid. Cytokeratin 5/6 staining was also present in 33/34 (97%) of mesothelioma cases. Six (9%) adenocarcinomas were positive, including metastases from the lung (1), breast (1), ovary (2) and unknown primary site (2). Four of the six adenocarcinoma cases positive for CK5/6 were in ascitic fluids. No cases of mesothelioma were negative for both calretinin and CK5/6. Only one adenocarcinoma case, (which was from unknown primary site in an ascitic fluid sample), was positive for both markers. Conclusions: The results confirm that calretinin and CK 5/6 are useful markers for mesothelioma in effusion specimens. CK5/6 staining may be less useful for peritoneal fluid specimens where metastatic adenocarcinomas may be more likely to express the antigen. Further study of ascitic/peritoneal specimens is warranted. However, positive staining, particularly for both antigens, is highly indicative of a mesothelial origin for cells. The two markers make a useful addition to EMA and the panel of adenocarcinoma markers routinely applied to effusion specimens.

Calretinin (CR) is a valuable marker in the immunohistochemical distinction between malignant mesothelioma (MM) and adenocarcinoma (ACA) in tissue sections. However, there is limited and conflicting data regarding the utility of CR in this differential diagnosis on cytologic material, especially cell block preparations. Also, the possible role of CR in the distinction of papillary serous borderline tumor (SBT) cells from reactive mesothelial cells in peritoneal washings has not been examined.Formalin-fixed paraffin-embedded cell block specimens of cytologic fluids, washings, and aspirates with a suspicious or positive cytologic diagnosis and a confirmed diagnosis of MM (29 cases), ACA (39 cases), and SBT (10 cases) were used for CR immunohistochemistry (IHC).Moderate to strong staining in50% of tumor cells was noted in 26 of 29 (90%) MMs but in only 3 of 39 (8%) ACAs and 1 of 10 (10%) SBTs. Admixed reactive mesothelial cells (when present) were strongly positive in all fluid specimens, but the staining pattern of benign non-reactive mesothelial cells in washing specimens was less reliable.CR is both a sensitive and specific marker of reactive and neoplastic mesothelial cells in cytologic cell block preparations and is thus useful in the differential diagnosis of MM and metastatic ACA in malignant effusions. However, CR IHC does not appear to allow definitive identification of SBT in peritoneal washings.

Context.—The cytologic distinction between pleomorphic adenoma (PA) and adenoid cystic carcinoma (ACC) can be diagnostically challenging in aspirate smears. Hence a cytologic diagnosis of “atypical cytology” with a differential diagnosis including PA and ACC is occasionally rendered in a subset of salivary gland fine-needle aspirations. Objective.—To evaluate the role of glial fibrillary acidic protein (GFAP) and CD57 expression in cell block material obtained during fine-needle aspiration procedure in differentiating PA from ACC. Design.—We performed GFAP and CD57 immunostains on formalin-fixed, paraffin-embedded cell block sections of 26 salivary gland fine-needle aspiration cases with the following cytologic diagnoses: (1) PA (10 cases); (2) atypical cytology, cannot exclude ACC (8 cases); and (3) ACC (8 cases). Results.—All 10 (100%) cases with cytologic diagnoses of PA were positive for GFAP, and 8 (80%) of 10 cases were positive for CD57; tissue follow-up confirmed the diagnosis of PA in all cases. All 8 (100%) cases with cytologic diagnosis of ACC were negative for both GFAP and CD57; tissue follow-up confirmed the diagnoses of ACC in all cases. Of the 8 cases with diagnoses of atypical cytology, 4 (50%) were negative and 4 (50%) were positive for both GFAP and CD57. Subsequent tissue follow-up in these cases revealed 4 cases of ACC (all negative for GFAP and CD57) and 4 cases of PA (all positive for GFAP and CD57). Conclusions.—Our results show that positive staining for GFAP and CD57 serves as a useful adjunct for the diagnosis of PA and helps to reduce the uncertainty in challenging cases.

Serous tumors of low malignant potential are uncommon in women under 30-years-old peritoneal washings play an important role in the diagnosis and prognosis of ovarian neoplasms. Accurate diagnosis of peritoneal washings is important in determining therapeutic regimens and in determining patient prognosis. In peritoneal washings, these tumors can be difficult to distinguish from reactive benign mesothelial cells. The cases of four women with borderline serous ovarian tumors, all of whom were under the age of 25, and had peritoneal washings which were positive for papillary tumors were reviewed.

We compared the results of smears to those of cytospin and cell block preparations from fine-needle aspirations to determine the cost effectiveness of each and to determine which should be routinely obtained. We reviewed 844 cases, 361 of which had both smears and cytospins, and 483 of which had both smears and cell blocks. Smears alone were diagnostic in 94% of cases (796/844 cases), cytospins alone diagnostic in 43% of cases (154/361 cases), and cell blocks alone diagnostic in 57% of cases (277/483 cases). Cytospins contributed additional information beyond that obtained from smears in 2% (7/361) and cell blocks in 12% (57/483) of cases. When smears were nondiagnostic, cytospins contributed additional information in 10% (2/21) of cases and cell blocks contributed additional information in 44% (12/27) of cases. The cost of providing a diagnosis from smear alone is $212, from cytospin alone is $352, and from cell block alone is $392. The cost for additional information established by cytospin is $7,736 and by cell block the cost is $1,906. Smears are superior to either cytospins or cell blocks in providing a diagnosis. It is not cost-effective to obtain either cytospins or cell blocks in addition to smears on all cases. However, it is cost-effective to obtain cell blocks when the immediate smear evaluation is nondiagnostic.

AIMS: To review consecutive cell block preparations of cytological specimens in a large general hospital. METHODS: 50 cell blocks were made over an 18 month period in which about 1900 fine needle aspirations (FNAs) were performed. The aspirator was a cytologist or, for image guided FNAs, a radiologist with a cytologist at hand to collect the specimen. Forty eight cell blocks were from FNAs and two were from serous fluids. RESULTS: The cellularity of the cell blocks was inadequate in only four preparations. The main motive for making cell blocks was to obtain tissue for immunohistochemistry. This was performed on 28 cases and a total of 107 immunostained sections were produced. The most common diagnostic dilemma was between carcinoma and melanoma, and the second most common between carcinoma and lymphoma. Consequently cytokeratin, S-100, and LCA were the most frequently used antibodies. At least one of these three antibodies was positive in 17 cases. Five cases were immunostained only for prognostic breast markers. CONCLUSIONS: The use of cell block immunohistochemistry is a reliable and technically unsophisticated aid in the cytological examination of tumours other than lymphomas. Success depends on having highly experienced aspirators that reliably obtain sufficiently cellular material.

Immunocytochemistry on fine needle aspiration (FNA) material has been mainly performed on cytologic preparations; there have been few reports on the use of FNA cell blocks. This study compared the intensity scores of immunoperoxidase staining on FNA cell block preparations from 21 breast, 12 thyroid and 10 lymph node aspirates with the scores on the corresponding surgically excised specimens. FNA materials for cell blocks were fixed in formalin and embedded in agar. Ten commercially available antibodies forming three panels were studied using standard peroxidase-antiperoxidase and avidin-biotin complex techniques. In general, the staining results on the FNA cell block sections agreed with those on the surgical specimens; in addition, there were fewer aberrant positive staining results and much less background staining in the cell block sections. These phenomena were most striking with the cytokeratin antibodies. It is concluded that immunoperoxidase staining on FNA cell block preparations is reliable; the advantages of the use of cell block sections as opposed to smears are discussed.

Background The development of high-throughput drug screening (HTS) using primary cultures provides a promising, clinically translatable approach to tailoring treatment strategies for patients with cancer. However, this has been challenging for solid tumors because of often limited amounts of tissue available. In most cases, in vitro expansion is required before HTS, which may lead to overgrowth and contamination by non-neoplastic cells. Methods In this study, hematoxylin and eosin staining and immunohistochemical staining were performed on 129 cytopathology cases from 95 patients. These cytopathology cases comprised cell block preparations derived from primary tumor specimens or patient-derived xenografts as part of a pediatric precision oncology trial. Cytopathology cases were compared with the morphology and immunohistochemical staining profile of the original tumor. Cases were reported as tumor cells present, equivocal, or tumor cells absent. The HTS results from cytopathologically validated cultures were incorporated into a multidisciplinary tumor board report issued to the treating clinician to guide clinical decision making. Results On cytopathologic examination, tumor cells were present in 77 of 129 cases (60%) and were absent in 38 of 129 cases (29%), whereas 14 of 129 cases (11%) were equivocal. Cultures that contained tumor cells resembled the tumors from which they were derived. Conclusions Cytopathologic examination of tumor cell block preparations is feasible and provides detailed morphologic characterization. Cytopathologic examination is essential for ensuring that samples submitted for HTS contain representative tumor cells and that in vitro drug sensitivity data are clinically translatable.

Pleural fluid is a liquid located in the pleural cavity, between the pleural visceral and pleural parietal. Analysis of pleural fluid is mandatory in all cases of diseases that give rise to pleural effusion. Cytopathological test gives the first indication of malignancy in malignant effusion, one of which is by making cell block. The stages of making cell block begin with fixation to maintain the cell component and its state is close to the original. The purpose of this study was to determine the comparison of the quality of the cytopathological examination results from the manufacture of cell block using a fixative solution of Nathan Alcohol Formalin Substitute (NAFS) and alcohol 96% -NBF 10% in pleural fluid samples. Experimental research with cross-sectional research design. The results showed that 13 samples of pleural fluid preparations in cell block preparations with NAFS fixative solution and cell block preparations with 96%-NBF 10% alcohol fixative solution there was no significant difference in the morphological figure of the cells, the number of cells produced, the number of malignant cells analyzed, the absorption of staining and cytodiagnosis sensitivity. Based on the results of this study, it can be concluded that alcohol 96% -NBF 10% can be used as an alternative solution to fixative for pleural fluid samples.

Introduction: Identifying metastatic breast carcinoma (mBC) in malignant effusion cytology (MEC) specimens is critical, as this will determine the patient’s prognosis and therapeutic management. Overlapping cytomorphologic features of breast carcinoma (BC) with other neoplastic entities makes the use of sensitive and specific markers highly desirable. Recent studies have reported trichorhinophalangeal syndrome type 1 (TRPS1) as a sensitive and specific marker for primary BC and mBC. We aimed to investigate TRPS1 expression in MEC of mBC and its most common diagnostic mimickers. Materials and Methods: A retrospective search from the pathology archives identified 82 MEC. TRPS1 expression in mBC was analyzed, and the results were compared to those in metastatic carcinoma of Müllerian origin (mMC) and metastatic pulmonary adenocarcinoma (mPAC). TRPS1 immunoperoxidase was performed on cytospin or cell block preparations, and p < 0.05 was considered significant. Results: Nuclear expression for TRPS1 was evaluated and scored as positive (≥1% of tumor cells) or negative. Nuclear TRPS1 expression was seen in 100% (30/30) mBC, 72% (18/25) mMC, and 7% (2/27) mPAC. This resulted in sensitivity, specificity, positive predictive value, and negative predictive values of 100%, 61%, 60%, and 100%, respectively. Conclusion: TRPS1 is a sensitive marker for mBC and can be reliably performed on cytology specimens. TRPS1 expression was also identified in a significant proportion of mMC, creating a potential diagnostic pitfall. Therefore, caution should be exercised when evaluating MEC of mBC with TRPS1. Consequently, a combination of immunoperoxidase panels should be employed in this setting.

Background: Fluid cytology plays an important role in delineating benign from malignant effusions, tumor staging, and also in diagnosing recurrences. Various methods are used in cytology for the preparation of smears. As the accurate diagnosis of the fluids aids in clinical decisions, the method of preparation of cytology smears, it is very important. Cytospin preparation of smears is one of the methods which provide higher cellular yield with better preservation of cellular morphology and is less time consuming. On the other hand, cell block method gives superior architectural details and provides options for immunocytochemistry. Aims and Objectives: The aim of this study was to assess the diagnostic utility of cytospin in comparison to cell block method in peritoneal and pleural fluid cytology. The study is done to determine the sensitivity, specificity, predictive values, and diagnostic accuracy of cytospin preparations with cell block method which is considered as the gold standard. Materials and Methods: This was a diagnostic test evaluation study done at the Department of Pathology, Government Medical College, Kottayam. The sample size was 240 which included all pleural and peritoneal fluids received in our cytology laboratory during the study period. Cytospin prepared smears of peritoneal and pleural fluids were compared with the tissue sections prepared by cell block method, to evaluate the diagnostic utility of cytospin. Tissue sections prepared from the cell blocks of effusions were considered as the gold standard for comparison. Results: A diagnostic test evaluation of cytospin preparation was done with cell block preparations. The sensitivity of cytopsin preparations in pleural and peritoneal fluid cytology is 94%. The specificity of cytopsin preparations in pleural and peritoneal fluid cytology is 100%. The positive predictive value of cytopsin preparations in pleural and peritoneal fluid cytology is 100% and the negative predictive value of cytopsin preparations in pleural and peritoneal fluid cytology is 96.8%. Hence, accuracy of the test is 97.9%. Conclusion: There is only minimal statistical difference between the results obtained by the cytospin and cell block methods. Cytospin method is less time consuming along with the advantage of higher cellular yield. Hence, the incorporation of cytospin along with the cell block technique is beneficial for augmenting the results of effusion cytology.

Introduction: Metastatic cancers are frequently detected on fine-needle aspiration (FNA) cytology, and confirmation of metastatic breast cancer often requires immunocytochemistry. Tissue provisioning for FNA specimens is important. In this study, GATA3, gross cystic disease fluid protein-15 (GCDFP15), mammaglobin (MMG), and SOX10 were performed on cell block preparations from aspirates of histologically confirmed metastatic breast cancers. The diagnostic performance of single markers and combinations of these markers were investigated with the aim to construct a tissue-efficient immunopanel. Methodology: Aspirates of metastatic breast cancer with corresponding histology and biomarker (estrogen receptor (ER), progesterone receptor (PR), HER2 and ki67) profile were retrieved. ER, GATA3, GCDFP15, MMG and SOX10 immunostains were performed on cell block sections and their expressions were assessed and compared. Results: Immunostaining was performed on a total of 115 aspirates. GATA3 showed the highest expression, followed by MMG, GCDFP15 and SOX10. Twenty-three, five and five cases expressed GATA3, MMG and SOX10 only. The five cases expressing SOX10 only were ER negative, and SOX10 expression was negatively associated with ER (p = 0.001), MMG (p = 0.001), GCDFP15 (p = 0.010) and GATA3 (p = 0.002), whereas GATA3 expression showed positive correlation with ER positivity (p < 0.001). MMG and GCDFP15 showed association with high Ki67 (p < 0.05), and no correlations were found with HER2 expression. Conclusion: In this cohort, GATA3 was the most sensitive single marker. The addition of MMG and SOX10 increases the sensitivity for detection of ER positive and ER negative breast cancers, respectively. These findings support the use of a combination of GATA3/MMG/SOX10 for confirmation of metastatic breast cancer.

Fine-needle aspiration cytology (FNAC) is generally the initial sampling method for salivary gland neoplasms. The cytomorphologic features of acinic cell carcinoma (AciCC) of salivary gland can overlap with other neoplastic and nonneoplastic entities. AciCCs harbor a recurrent t(4;9) rearrangement with upregulation of nuclear receptor subfamily 4 group A member 3 (NR4A3). NR4A3 protein overexpression has been shown to be highly sensitive and specific for the diagnosis of AciCC in histologic specimens and cell block preparations. However, data on NR4A3 immunocytochemistry (ICC) on conventional smears or liquid-based cytology are limited.The authors identified 18 FNAC of histologically proven AciCC cases between 2013 and 2019. FNAC samples of diagnostic mimickers were likewise retrieved and included in the study cohort for comparison. The NOR1/NR4A3 mouse monoclonal antibody was applied directly to FNAC slides using a standard ICC technique.The cohort included ethanol-fixed Papanicolaou-stained cytologic smears and liquid-based preparations from 18 AciCC, one secretory carcinoma, four mucoepidermoid carcinomas, four salivary duct carcinomas, five pleomorphic adenomas (PA), five Warthin tumors, five oncocytomas, one oncocytic hyperplasia, and five nonneoplastic salivary gland cases. Strong nuclear staining for NR4A3 was present in all AciCC, weak nuclear staining was present in one PA, and all other non-AciCC were negative (sensitivity, 100%; specificity, 97%).NR4A3 ICC can be used directly on FNAC conventional smears and liquid-based cytology to reliably distinguish AciCC from its mimickers. This marker may be useful in cases where a cell block preparation is unavailable or inadequate.