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The utility of cell block (CB) preparation is well established in cytopathology. Despite 23.3% of College of American Pathologists–accredited laboratories using CB with liquid-based preparations on urine cytology (UC) cases, there are very few studies on their performance. To determine their usefulness, we conducted a retrospective review of UC cases that received CB. We identified 27 UC cases with ThinPrep (TP) and CB preparation between 2016 and 2020 at our institution. Clinical history and follow-up data were compiled. A blinded review of TP alone, and TP together with CB, was performed by 2 pathologists and 2 cytotechnologists. Diagnoses were rendered in accordance with The Paris System for Reporting Urine Cytology. Blood and acute inflammation were common background elements in cases that received CB preparation. In total, CB upgraded the diagnosis in 7 of 27 cases (26%). The maximum utility of CB preparation was seen in indeterminate cases where 60% (6 of 10) were upgraded, including 71% (5 of 7) of atypical urothelial cells (AUC) and 30% (1 of 3) of suspicious for high-grade urothelial carcinoma (HGUC). One case (1 of 12, 8%) diagnosed as negative for HGUC on TP was diagnosed as low-grade urothelial neoplasia on CB. Our results demonstrate that adjunct use of CB preparation aids in a definitive diagnosis on AUC category and may be helpful in cases with cell clusters or tissue fragments, or cases suspicious for HGUC. Further correlation studies are warranted in this area to expand our knowledge about the utility of CBs in urine cytology. • More studies are needed on cell block preparations on urine cytology specimens. • Indeterminate urine cytology cases show the most benefit of cell block preparation. • We highlight many areas for further research on cell blocks in urine cytology. [ABSTRACT FROM AUTHOR]

Introduction: A palpable breast lump is a common diagnostic problem for clinicians and surgeons. Fine-needle aspiration cytology (FNAC) has many advantages such as less cost, less sample processing time, less pain, less chance of hematoma, and less discomfort. FNAC with cell block preparation further increased both sensitivity and specificity by nearly 100%. With the cell block preparation, we can also use newer tests like estrogen receptor-progesterone receptor-human epidermal growth factor receptor 2., Aims: The aim of this study was to derive conclusions about the correlation, including sensitivity, specificity, positive and negative predictive values (NPVs), and the diagnostic accuracy of FNAC, with or without cell blocks, compared to the final histopathology in cases of palpable breast masses., Materials and Methods: A cross-sectional prospective study was conducted after getting approval from the Human Ethics Research Committee from January 2018 to December 2019, which included 65 patients. Patients diagnosed clinically for breast lumps who underwent diagnostic FNAC with cell block, followed by a histopathological examination at our hospital, were included in the study., Results: FNAC without cell block sensitivity, specificity, positive predictive value (PPV), NPV, efficiency rate, and diagnostic accuracy are 91.3%, 100%, 100%, 90.1%, 86.2%, and 96.5%, respectively. FNAC with cell block sensitivity, specificity, PPV, NPV, efficiency rate, and diagnostic accuracy are all 100%. All of our results beat the standard estimate., Conclusions: Fine-needle aspiration cytology is a patient-friendly, easy, reliable, repeatable, and simple diagnostic test. Whenever it is combined with cell block preparation, improves the accuracy of FNAC diagnosis which is more accurate and comparable to golden-standard biopsy with histopathology examination., Competing Interests: There are no conflicts of interest., (Copyright: © 2024 Journal of Mid-life Health.)

Background: PD-L1 immunoexpression in head and neck squamous-cell carcinoma (HNSCC) determines immunotherapy eligibility. Patients are often diagnosed using fine-needle aspiration (FNA) of metastatic lymph nodes, however, the cytohistologic correlation of the combined positive score (CPS) is largely unknown., Methods: This study retrospectively identified 96 paired histologic (HS) and cytologic specimens (CyS), between 2016 and 2020, diagnosed with HNSCC. Cases with <100 tumor cells (n = 54) or missing block(s) (n = 8) were excluded. All 34 case pairs were scored with CPS using the PD-L1 22C3 pharmDx assay at clinically relevant cut-offs of <1%, 1%-19%, and ≥20% independently by three observers blinded to the case pairs (CyS with corresponding HS)., Results: The CPS (<1/1-19/≥20) for CyS and HS were as follows: 10(29.4%)/10(29.4%)/14(41.2%) and 2(5.9%)/13(38.2%)/19(55.9%), respectively. There was fair overall cytohistologic agreement (OA) of 76.5% (k = 0.261) at the CPS cut-off of 1. The OA did not differ significantly between site-matched (n = 13) and -unmatched (n = 21) case pairs (p = .4653). CyS has a specificity and positive predictive value (PPV) of 100% but a negative predictive value (NPV) of only 20% as compared to its paired HS., Conclusions: Our study demonstrates fair CPS cytohistologic correlation in HNSCC specimens using the PD-L1 IHC 22C3 pharmDx assay with high PPV but low NPV. This suggest that determining PD-L1 status in FNA specimens can play an important role in the clinical management of HNSCC patients., (© 2023 The Authors. Diagnostic Cytopathology published by Wiley Periodicals LLC.)

Abstract Circulating tumor cells have emerged as biomarkers for estimating the tumor burden and metastatic potential of cancer patients. However, to date, most of studies and applications of circulating tumor cells have been conducted and applied to epithelial cancers such as breast, colorectal, and prostate tumor. The only FDA-cleared method, CellSearch, makes use of antibody against epithelial surface protein expressed on CTCs, thus obstructing wide application for various cancers with non-epithelial and semi-epithelial characteristics including renal cell carcinoma. Due to rarity and ambiguity of CTCs, designed experiment including non-biased CTC isolation and subsequent cytopathological study for finding applicable immunomarkers are urgently needed for clinical use of CTCs for less-studied cancers. Here, in order to construct the fundamental step for CTC diagnosis without limitation of its epithelial characteristics, we present the simple and novel method which incorporate both label-free CTC isolation and pathological study using hydrogel-based cell block formation. Six cell lines from lung, ovarian, kidney cancers were used to make cell block and analyzed by conventional immunocytochemical staining method to find the candidate markers for CTC. Especially for renal cancer, the physically isolated CTCs were further immunocytochemically examined with the screened candidate markers by cell block construction, and verified their clinical utility using blood samples from patients with renal cell carcinoma. This comprehensive study demonstrates that the present approach can be used to find the potential markers for any type of cancers regardless of their epithelial characteristics and isolate the specific type of CTCs in label-free manners.

Background& Objective: Fine needle aspiration cytology (FNAC) is an easy, rapid, and less hazardous tool to diagnose the intra-abdominal lesions with various imaging modalities adding to its sensitivity and accuracy. However, sometimes it does not yield adequate information for precise diagnosis and the risk of false-negative and indeterminate diagnosis is always present. Cellblock preparations may be particularly helpful in such problematic cases. The current study aimed at evaluating and comparing the cytological as well as histopathological features of different intra-abdominal mass lesions. Methods: Image-guided FNAC followed by cell block were performed on 167 patients from June 2012 to May 2013. Histologically correlated 111 cases were evaluated. Results of conventional smear, cell block, and combination of FNAC with cell block were compared with histopathological findings regarding diagnostic sensitivity, specificity, and accuracy of diagnosis. Result: Cell block was more specific to diagnose these lesions than FNAC (95.49% versus 90.09%). Combined application of cell block with FNAC was more specific (96.39%) than cell block alone with 100% diagnostic accuracy. Conclusion: Application of a combination of cell block with FNAC was more useful to diagnose intra-abdominal mass lesions.

Background: Cytologic examination of body fluids commonly involves the use of direct or sediment smears, cytocentrifuge preparations, membrane filter preparations, or cell block sections. Cytospin and cell block techniques are extremely useful in improving cell yield of thin serous effusions and urine samples, and ensure high diagnostic efficacy. Materials and Methods: We studied cytospin preparations and cell block sections prepared from 180 samples of body fluids and urine samples to compare the relative efficiency of cell retrieval, preservation of cell morphology, ease of application of special stains, and diagnostic efficacy. Samples were collected and processed to prepare cytospin smears and cell block sections. Results: We observed that overall, cell yield and preservation of individual cell morphology were better in cytospin preparations as compared to cell blocks, while preservation of architectural pattern was better in cell block sections. The number of suspicious cases also decreased on cell block sections, with increased detection of malignancy. It was difficult to prepare cell blocks from urine samples due to low cellularity. Conclusions: Cytospin technology is a quick, efficient, and cost-effective method of increasing cell yield in hypocellular samples, with better preservation of cell morphology. Cell blocks are better prepared from high cellularity fluids; however, tissue architecture is better studied, with improved rate of diagnosis and decrease in ambiguous results. Numerous sections can be prepared from a small amount of material. Special stains and immunochemical stains can be easily applied to cell blocks. It also provides a source of archival material.

Background: Immunohistochemistry (IHC) is a routinely performed method to demonstrate estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) in surgical breast cancer specimens but not on cell block (CB) of fine-needle aspiration (FNA). The aims of this study were to evaluate the expression of ER, PR, and HER2 using immunocytochemistry (ICC) on CB and compare with the corresponding tissue blocks as gold standard as well as to compare with other similar studies. Materials and Methods: Forty-eight breast carcinoma CB specimens with their corresponding tissue blocks were identified. ICC on CB for ER, PR, and HER2 was performed and compared with tissue blocks. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value were measured for each receptor. The degree of agreement between CB and tissue blocks was calculated using Cohen's kappa (κ) test. Results: ER results showed 67.7% sensitivity, 94.1% specificity, 95.5% PPV, and a moderate agreement (κ =0.588). PR results showed 50% sensitivity, 90% specificity, 87.5% PPV, and a fair agreement (κ =0.368). HER2 results showed 58.3% sensitivity, 100% specificity, 100% PPV, and a moderate agreement (κ =0.539). Conclusion: The results of this study confirm the wide variations that occur between CB ICC and tissue block IHC in the detection of ER, PR, and HER2 in breast cancers. In comparison with other studies, we report a low sensitivity and high specificity rates for ER, PR, and HER2 in FNA CB. Further studies are recommended.

Objectives: Recent investigations have shown strong correlations between cytology and surgical non-small cell lung carcinoma (NSCLC) specimens in programmed death-ligand 1 (PD-L1) immunohistochemical (IHC) evaluations. Our study aims to evaluate the reproducibility of PD-L1 IHC scoring in NSCLC cytology cell blocks (CBs) and to assess the impact of CB cellularity, method of sample collection, and observer subspecialty on scoring agreement.Methods: PD-L1 IHC was performed on 54 NSCLC cytology CBs and was scored independently by seven cytopathologists (three of seven with expertise in pulmonary pathology). Three-tier scoring of negative (<1%), low positive (1%-49%), and high positive (≥50%) and interrater agreement were assessed.Results: Total and majority agreement among cytopathologists was achieved in 48% and 98% of cases, respectively, with κ = 0.608 (substantial agreement; 95% confidence interval, 0.50-0.72). Cytopathologists with pulmonary pathology expertise agreed in 67% of cases (κ = 0.633, substantial agreement), whereas the remaining cytopathologists agreed in 56% of cases (κ = 0.62, substantial agreement). CB cellularity (P = .36) and sample collection type (P = .59) had no statistically significant difference between raters.Conclusions: There is substantial agreement in PD-L1 IHC scoring in cytology CB specimens among cytopathologists. Additional expertise in pulmonary pathology, sample collection type, and CB cellularity have no statistically significant impact on interobserver agreement. [ABSTRACT FROM AUTHOR]

BRCA1‐associated protein 1 (BAP1) or methylthioadenosine phosphorylase (MTAP) immunohistochemistry (IHC) or 9p21 fluorescence in situ hybridization (FISH) are useful for the diagnosis of malignant pleural mesothelioma (MPM). However, the effect of these assays on the diagnostic yield of effusion cytology in MPM cases with suspicious cytomorphology or the diagnostic challenges in BAP1 or MTAP IHC have not been fully elucidated. Two cohorts of cytologic preparations obtained from pleural effusions were examined: MPM cases in cohort 1 were used to evaluate whether BAP1 or MTAP IHC or 9p21 FISH increase the diagnostic yield of effusion cytology; cohort 2 included cases suspicious for MPM, to which BAP1 or MTAP IHC was applied to clarify the challenges in the clinical assessment of these assays. In cohort 1 (n = 28), either assay elevated 62.5% of class II or III cases to class V. In cohort 2 (n = 139), 21.7% of BAP1 immunocytochemistry in smears and 10.6% of BAP1 IHC and 9.4% of MTAP IHC in cell blocks, were identified to be challenging. The application of genomic‐based assays increased the diagnostic yield of effusion cytology in the diagnosis of MPM. However, diagnostic challenges limit the application of these assays in some cases. [ABSTRACT FROM AUTHOR]

Background: Cell-free DNA (cfDNA) is emerging as a surrogate sample type for mutation analyses. We investigated the suitability of malignant pleural effusion (MPE) and plasma as a biomaterial for analyzing epidermal growth factor receptor (EGFR) mutation by peptide nucleic acid (PNA) clamping-assisted fluorescence melting curve (PANAMutyper™) analysis. Methods: Matched tissue, MPE cell block (MPE-CB), MPE supernatant, and plasma samples were collected from patients with advanced lung adenocarcinoma who had a MPE at the time of diagnosis. EGFR mutation was assessed by PANAMutyper™. Results: Mutation analyses in matched tumor tissues, MPE-CB, MPE supernatant, and/or plasma samples were available for 67 patients. In comparison with tumor tissue and MPE-CB, MPE supernatant exhibited 84.4% sensitivity, 97.1% specificity, 96.4% positive predictive value (PPV), and 87.2% negative predictive value (NPV). In the same comparison, plasma exhibited 70.6% sensitivity, 100.0% specificity, 100.0% PPV, and 73.7% NPV. When sorted by mutation type, MPE supernatant had better sensitivity than plasma for the detection of two major EGFR mutations: 93.8% vs. 75.0% for exon 19 deletion and 73.3% vs. 60.0% for L858R. Conclusions: In this cohort of patients with MPEs, MPE supernatant demonstrated superior diagnostic performance compared with plasma using a PNA-based real-time PCR method. [ABSTRACT FROM AUTHOR]

Background: The Ki‐67 index is important for grading neuroendocrine tumors (NETs) in cytology. However, different counting methods exist. Recently, augmented reality microscopy (ARM) has enabled real‐time image analysis using glass slides. The objective of the current study was to compare different traditional Ki‐67 scoring methods in cell block material with newer methods such as ARM. Methods: Ki‐67 immunostained slides from 50 NETs of varying grades were retrieved (39 from the pancreas and 11 metastases). Methods with which to quantify the Ki‐67 index in up to 3 hot spots included: 1) "eyeball" estimation (EE); 2) printed image manual counting (PIMC); 3) ARM with live image analysis; and 4) image analysis using whole‐slide images (WSI) (field of view [FOV] and the entire slide). Results: The Ki‐67 index obtained using the different methods varied. The pairwise kappa results varied from no agreement for image analysis using digital image analysis WSI (FOV) and histology to near‐perfect agreement for ARM and PIMC. Using surgical pathology as the gold standard, the EE method was found to have the highest concordance rate (84.2%), followed by WSI analysis of the entire slide (73.7%) and then both the ARM and PIMC methods (63.2% for both). The PIMC method was the most time‐consuming whereas image analysis using WSI (FOV) was the fastest method followed by ARM. Conclusions: The Ki‐67 index for NETs in cell block material varied by the method used for scoring, which may affect grade. PIMC was the most time‐consuming method, and EE had the highest concordance rate. Although real‐time automated counting using image analysis demonstrated inaccuracies, ARM streamlined and hastened the task of Ki‐67 quantification in NETs. The Ki‐67 proliferation index in cell blocks of neuroendocrine tumors typically is performed in a variety of ways at different institutions. The current study compares traditional and novel scoring methods for Ki‐67 enumeration in cytology cell blocks. The results indicate that augmented reality microscopy coupled with real‐time image analysis can simplify and hasten this task. [ABSTRACT FROM AUTHOR]

Background and Aims: Advent of personalised treatment needs correct diagnosis of lung adenocarcinoma with its molecular subtyping. Minimal use of special stain or immunohistochemistry (IHC) in small specimens save material for molecular testing. Various histologic patterns in adenocarcinoma (ADC) subtypes have different prognostic implications and current recommendation is to describe these patterns in small specimens. Aim of this study was to diagnose adenocarcinoma from cytology specimens depending on adenocarcinoma pattern on fine needle aspiration smears and cell blocks. We also studied the additional role of cell blocks as a platform for special stain and IHC. Materials and Methods: Conventional smears and cell block (CB) preparation were examined from transthoracic CT guided FNA samples of suspicious lung malignancy cases. Clear defining architectural pattern and cytomorphological features in favour of adenocarcinoma were evaluated and mucin stain and IHC were used as and when required. Results: A total of 86 cases were included in this study, of which 83 cases were diagnosed as adenocarcinoma, 52 (62.5%) showed clear cut evidence of adenocarcinoma from smears and CBs. CB morphology alone aided the diagnosis in 12. Various ADC patterns in combination or alone were appreciated in these 64 cases. Sixteen needed mucin stain and 3 needed IHC for diagnosis. Forty one were ADC with solid pattern of which 39 showed high nuclear grade. Conclusion: Adequately cellular FNA smears and corresponding cell blocks of optimal quality can aid effectively in diagnosing adenocarcinoma and appreciating its pattern. Therefore, it would minimize the need for special stain and/or IHC with preservation of more material for molecular testing. [ABSTRACT FROM AUTHOR]

Background: Diagnosis of lung cancer can be made in two ways: histopathological and cytopathological. Cytological methods in the diagnosis of lung lesions are generally thought to be one of the most successful tactics. Aims: This study aimed at comparing the efficiency of selected cytological techniques in lung lesions by correlating them with histopathological diagnosis. In addition, we had answered the question whether any of the cytological methods can replace histopathology. Materials and Methods: The study group consisted of 633 patients and 1085 cytological specimens. Cytology samples included: induced sputum, bronchial washing (BW), bronchial brushing (BB), fine needle aspiration (FNA), and cell block (CB). In every case of CB immunocytochemistry (ICC) was performed. For each cytological method sensitivity, specificity, effectiveness, positive predictive value, and negative predictive value were assessed. Results: BW and BB showed the lowest diagnostic parameters. The most valuable diagnostic procedure was CB based on FNA. Close by CB, FNA had the highest diagnostic rate. However, possibility to evaluate tumor cell structure and apply the ICC, give CB an advantage over FNA. Using only morphologic criteria, we had subclassified nonsmall-cell lung cancer (NSCLC) into squamous cell carcinoma (SCC) and adenocarcinoma (AC) as 60.04% of SCC and 32.52% of AC. The use of CB and ICC decreased the NSCLC diagnoses from 22.1% to 2.8% while the percentage of AC and SCC diagnoses increased from 4.11% to 12.64% and from 6.64% to 11.06%, respectively. Metastatic lung tumors were diagnosed based on both the cell morphology and according to the ICC results. Conclusion: Despite the limitations of the cytological procedures, we recommend using CB and ICC to evaluate cytological samples derived from FNA. It can in many cases replace a conventional histopathology. [ABSTRACT FROM AUTHOR]

Primary hepatoid adenocarcinoma (HAC) is a rare extrahepatic adenocarcinoma with morphological and phenotypical resemblance to hepatocellular carcinoma (HCC). It can occur in lung, stomach, gallbladder, pancreas, ovary, and uterus, with most common site being stomach. Morphological features of primary HAC of the lung are similar to HCC, so exclusion of metastatic HCC is necessary. In this report, we describe two cases of elderly men with primary pulmonary HAC who presented in advanced clinical stage and diagnosed by fine-needle aspiration cytology with immunohistochemistry. Both patients succumbed to death despite starting first cycle of chemotherapy. [ABSTRACT FROM AUTHOR]

Background: Diagnostic cytology is study of cells that are exfoliated from epithelial surface or remove from the tissue. Cytological examination of body fluid helps us to differentiate inflammatory effusion from malignant effusion. It has diagnostic, prognostic and therapeutic implications. The method has disadvantage of lower sensitivity to detected and differentiate malignant cell from reactive atypical mesothelial cells. Aims: The aim of this study is to compare the cytological features of conventional smear (CS) method and cell block (CB) method in serous effusions and access their ability to diagnose malignancy. Material And Methods: Nighty seven serous effusion (pleural and peritoneal) samples were subjected to evaluation by both CS and CB methods. Each fluid specimen was divided into two equal parts, Cs and CB prepared. Cell blocks were prepared using plasma-thromboplastin cell block technique. Cellularity, architecture patterns, morphological features and yield for malignancy were compared, using the two methods. Result: Among 97 cases common age was 41-60 year with M:F sex ratio 1.06:1. Most common cause of effusion were Pulmonary tuberculosis (20.69%) followed by Ovarian mass (19.59%), Liver cirrhosis (6.19%), Lung mass (5.15%).Cell block was diagnostically superior (Score 6-8) in 34.02% cases whereas no cases were found diagnostically superior with conventional smear. The additional yield of malignancy was found to be 11.94% more by cell block method compared to conventional smear. Conclusion: The CB method provides high cellularity, better architectural patterns, morphological features and an additional yield of malignant cells, and thereby, increases the sensitivity of the cytodiagnosis when compared with the CS method. [ABSTRACT FROM AUTHOR]

Endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) is presently the preferred modality for sampling mediastinal lymph nodes. There is an unmet need for standardization of processing and reporting of cytology specimens obtained by EBUS-TBNA. The manuscript is a state-of-the-art review on the technical aspects of processing and reporting of EBUS-TBNA specimens. A literature search was conducted using the PubMed database, and the available evidence was discussed among the authors. The evidence suggests that at least one air-dried and one alcohol-fixed slide should be prepared from each lymph node pass. The remaining material should be utilized for microbiological analysis (in saline) and cell block preparation (10% formalin or other solutions). Wherever available, rapid-onsite evaluation should be performed to assess the adequacy of the sample and guide the need for additional material. The lymph node aspirate should also be collected in Roswell Park Memorial Institute solution in cases where lymphoma is under consideration. The use of liquid-based cytology provides good quality specimens that are free from blood and air-drying artifacts and can be used wherever available. Sample adequacy and the diagnostic category should be furnished separately in the cytology report. [ABSTRACT FROM AUTHOR]

Background: Being a minimally invasive diagnostic technique, Fine-Needle Aspiration Cytology (FNAC) has become the first-line test and corresponding aspirated material has become the target specimen for diagnosis and ancillary tests in lung carcinoma. Although the role of Cell Blocks (CBs) in diagnosis and in ancillary testing is well recognized in literature, limited attention has been paid to specimen procurement and triage in the preparation of CBs. In the present scenario, CBs are not consistently optimal because of its low cellularity. Aims: This study is aimed to describe an improvised technique of specimen acquisition and cell block preparation in CT-guided FNACs of lung carcinoma cases in a resource-constrained center and to assess its efficacy for optimal representation of cellularity, morphology, and architecture. Materials and Methods: Total 85 lung carcinoma cases undergoing CT-guided FNAC in our center from February 2017 to January 2018 were included in this study. 4 to 5 direct smears and subsequent CBs were made from material obtained by single pass. Cellularity of smears and corresponding cell blocks were assessed and categorized according to a scoring system (score 1 to 3 for number of cells <50, 50–100, >100, respectively). Preserved architecture and morphology were also assessed in smears and CBs. Results: The evaluated samples showed a cellularity score 3 in 65.4%CBs and score 2 in 24.7% CBs. Overall, 90.1% cell blocks had acceptable cellularity. Cell morphology was preserved in all CBs of acceptable cellularity, except for two adenocarcinoma, one squamous cell carcinoma, and one small cell carcinoma blocks. Cellular architecture was also preserved in all CBs of acceptable cellularity. Conclusions: This simple improvised technique of CB preparation optimized its cellularity, morphology, and architectural preservation, even after adequate cellular FNA smears. [ABSTRACT FROM AUTHOR]

• Treatment of ovarian malignant lymphoma is based on chemotherapy, although radical surgery improves prognosis of epithelial ovarian cancer. • Immunohistochemical analysis using cell block technology helped us establish an accurate diagnosis of ovarian lymphoma. • Abdominocentesis followed by cell block analysis is very useful for identifying ovarian malignancy before radical invasive operation. Ovarian malignant lymphoma is a rare gynecologic disease and some patients show marked ascites, similar to that observed in advanced ovarian cancer. Although radical surgery improves prognosis of ovarian cancer, treatment of lymphoma is based on chemotherapy, therefore, differential diagnosis is crucial. A 65-year-old woman presented with a 1-month history of abdominal distention. Pelvic ultrasonography showed an 11-cm solid mass in the pelvis. Computed tomography and magnetic resonance imaging revealed bilateral (mainly left) ovarian masses in the pelvis and multiple metastases. Laboratory examination revealed that serum CA125 levels were elevated, suggesting the existence of advanced ovarian cancer. To confirm the diagnosis, the ascites was removed via abdominocentesis. Although no malignant epithelial cells were observed, atypical lymphoid cells dispersed in the ascites were detected in the cytological analyses. Thus, for accurate diagnosis, we performed re-abdominocentesis and immunohistochemical (IHC) analysis using cell block technique. Cell block analysis showed negative staining for CD3 and positive staining for CD20 in large atypical lymphoid cells, suggesting the existence of large B-cell lymphoma. Repeat blood examination showed that the serum sIL-2R level was elevated. We decided to perform biopsy to make the final treatment decision. Histologically, the tumor demonstrated diffuse proliferation of large atypical lymphoid cells. IHC analysis showed CD3(-), CD5(+), and CD20(+). In addition, IHC analysis also showed CD79a(+), CD10(-), bcl-2(+), and cyclin D1(-). The final diagnosis was diffuse large B-cell lymphoma. Here, we present the case of a patient with ovarian malignant lymphoma that was diagnosed using cell block analysis. [ABSTRACT FROM AUTHOR]

Background: A peritheliomatous pattern (PP) in tumors is characterized by a sheath of viable tumor cells closely surrounding a central blood vessel. In the authors' cytology practice, such a PP has been recognized in several metastatic melanoma specimens. The aim of this study was to evaluate the frequency of a PP in cytology samples of melanoma in comparison with other tumors. Methods: Eighty archival fine‐needle aspiration (FNA) cases of metastatic melanoma were compared with 65 control cases (35 poorly differentiated/metastatic carcinomas, 15 lymphomas, and 15 recurrent/metastatic/high‐grade sarcomas). Cytologic findings were correlated with corresponding histologic specimens, which were available for 44 cases (55%) in the melanoma group and for 23 cases (35.38%) in the control group. All slides were examined for a PP and were semiquantitatively graded for comparison. Results: A PP was present in 51.3% of the cytologic preparations (n = 41) among the melanoma group cases, whereas in the control group, a PP was present in only 3.1% of the cases (n = 2). A PP was present in 65.9% of melanomas with available histologic sections (n = 29) and in 8.7% of tissue samples from the control group (n = 2). A PP was seen more often in cell blocks than direct smear preparations (51.3% vs 40.0%) from patients with melanoma. Conclusions: The presence of a characteristic PP can be helpful in diagnosing melanoma in FNA samples because it was present in almost half of the metastatic melanoma cases in this study and was rarely present in other tumor types. A PP is easier to recognize and more often presents in cell blocks than aspirate smears. Ancillary studies such as immunohistochemistry are helpful for excluding other entities that may exhibit a PP. In this study, the presence of a peritheliomatous pattern of tumor cells is shown to be helpful in diagnosing melanoma in fine‐needle aspiration smears and cell block preparations. This characteristic growth pattern is present in almost half of the metastatic melanoma cases in this study, and this highlights it as an important morphologic feature to recognize in fine‐needle aspirates of melanoma. [ABSTRACT FROM AUTHOR]

Aim: This study aims at investigating the concordance of programmed death-ligand 1 (PD-L1) expression in non-small cell cancer tissues that have been sampled using different methods and its relationship with the pathologic parameters of tumors. Methods: PD-L1 expression assays were made on the cell blocks taken using fine needle aspiration, the small tissue samples representing endoscopic biopsy and the large tissue samples representing the resected tumor taken from the tumors of 100 subjects diagnosed with non-small cell lung cancer; their percentage values were evaluated and their groups were determined based on these values. Results: The average difference in expression rates found through different sampling methods was close to 0, and the values of such differences changed mostly in a narrow range. In paired comparisons, a good level of concordance was observed between all the sampling methods. The values were 0.8865 (SE: 0.0306, CI: 0.8264-0.9466) in the comparison of tumor tissue (TT) and small biopsy (SB): 0.8637, (SE: 0.033, CI: 0.7989- 0.9285) TT to cell block (CB): 0.8916, (SE: 0.0272, CI: 0.8383- 0.9449) SB tissue to the CB. Conclusion: Therefore, it can be concluded that the PD-L1 expression does not differ between the tissues sampled through various methods in non-small cell lung cancers. [ABSTRACT FROM AUTHOR]

Cytological examination plays an important role in the initial work-up of the serous cavity effusion fluids to find out the possible etiology as benign or malignant. Among malignant effusions, cytology is helpful in determining the exact type, site, and stage of the tumor. However, for reporting effusion cytology specimens, there is no consistent and reproducible reporting system. Aims: The aim of these guidelines is to provide a standardized format for effusion cytopathology right from sample receipt to its ultimate report sign-out for implementation in all cytopathology laboratories. The Indian Academy of Cytologists in consultation with experts across the country has prepared guidelines pertaining to collection, preparation, and diagnostic categories of effusion specimens to reduce reporting variability. The guidelines are made keeping in mind the different areas of practices in India, especially low- and medium-resource settings. The guidelines are broadly divided into essential, optimal, and optional categories for best usage and appropriate allocation of the precious specimens. In referral centers or well-established setups, essential ancillary techniques can be done for accurate and final diagnosis. By adhering to and implementing these uniform guidelines, it is hoped that clinical patient care and management in India will improve and be of uniformly good quality by enabling and facilitating good laboratory practices. [ABSTRACT FROM AUTHOR]

Objective: To investigate the diagnostic utility of fine needle aspiration (FNA) smear, cell block (CB), flow cytometry (FC) and tuberculosis polymerase chain reaction (Tb‐PCR) analysis for Kikuchi's disease (KD). Methods: A total of 173 FNA biopsy samples were collected using a Youyi aspirator. KD was diagnosed by FNA smear, with or without, CB, FC and Tb‐PCR. Results: Out of 173 patients, 131 (75.7%) were female and 126 (72.8%) aged 21‐40 years. Of these, 171 patients (98.8%) presented with painful enlarged cervical lymph nodes. All cytological samples identified intracellular apoptotic debris embedded in the cytoplasm of crescentic and phagocytic macrophages. In 24 cell‐blocks (CBs), clusters of CD123 positive plasmacytoid monocytes were observed in KD. FC and CBs showed non‐specific lymphoid hyperplasia in nine cases with suspicious lymphoma. Tb‐PCR was negative in five cases with suspicion of tuberculosis. Conclusions: In summary, FNA biopsy is a fast, reliable, and relatively inexpensive diagnosis tool for KD. CB preparation is an important adjunct method for the diagnosis of KD. The authors investigated the diagnostic utility of fine needle aspiration (FNA) smear, cell block (CB), flow cytometry (FC) and tuberculosis PCR (Tb‐PCR) analysis for Kikuchi's disease (KD). They found that FNA biopsy allows cytopathologists to result in a fast, reliable, and relatively inexpensive diagnosis for KD. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to analyze immunohistochemical and immunocytological findings by examining enucleated eyes and vitreous cell block (CB) in patients with vitreoretinal lymphoma (VRL).Methods: Histological specimens were obtained from two enucleated eyes with VRL associated with neovascular glaucoma. CB specimens were prepared in 18 patients from diluted waste fluids containing shredded vitreous. Histological and cytological specimens were submitted for hematoxylin-eosin staining and immunopathological analyses.Results: Both specimens demonstrated massive infiltration of large lymphoma cells. The lymphoma cells were positive for CD20 and MUM-1 in enucleated eyes. Membranous immunoreactivity for CD20 was observed in lymphoma cells in CB with VRL. Bcl-6 and MUM-1 were marked in five and eight out of nine cases examined, respectivelyConclusions: Cytological findings in CB specimens indicated similar histopathological characteristics of enucleated eyes. CB specimens obtained from vitreous waste diluted fluids may serve as effective materials for cytological diagnosis of VRL. [ABSTRACT FROM AUTHOR]

We performed a comparison of cell blocks prepared with the collodion bag and HistoGel to evaluate the ease of embedding and cutting, performance with low cellularity specimens, time and cost per specimen, and value to support immunohistochemistry and molecular diagnostics. We processed 11 fresh, unfixed effusions using both the collodion bag and the HistoGel cell block preparation methods. Six immunohistochemistry stains were tested on 2 of the body fluids. DNA was extracted and quantified, and polymerase chain reaction cycle thresholds were evaluated from cell blocks prepared from 5 of the body fluids. The comparison parameters included embedding difficulty, cutting resistance, adequacy, cell yield, cell preservation, immunohistochemistry stain quality, DNA quantity, integrity, and purity. The time and cost to prepare each specimen was compared using normalized values for preparation of specimen, cost per year, and cost per specimen. Each parameter was assessed for both cell block preparation methods. All 3 of the samples with moderate or poor cell yield were low-volume (5-mL) samples prepared with the HistoGel method. In contrast, the collodion bag method produced a good yield with all three 5-mL samples. DNA recovery was greater in the collodion bag method. Similar crossing threshold values in purity reactions indicated equally high-quality matrix properties for the collodion bag and HistoGel preparations. Preparation of the specimen was 10 minutes faster with the collodion bag method, and the cost for the collodion bag method was $0.24 more expensive per cell block than using the HistoGel. The collodion bag method produced superior cell blocks for both morphologic and molecular studies more consistently, with lower volume specimens and with less time per specimen. [ABSTRACT FROM AUTHOR]

Objective: To compare the malignant cell detection rate between the conventional smear and the cell block method in ascites or peritoneal washing in endometrial and ovarian cancer. Materials and Methods: A cross-sectional study was conducted in 65 endometrial and presumed ovarian cancer patients who underwent surgical staging between August 2017 and February 2018 at Ramathibodi Hospital. All ascites or peritoneal washing fluids were examined by conventional smear and cell block method to detect malignant cells. The results were interpreted and reviewed by two cytologists. If the results were inconsistent or suspicious, re-interpretation and immunohistochemistry dyes were used to finalize the results. Results: Sixty-five specimens were examined. Conventional smear revealed negative for malignancy in 57 specimens and positive for malignancy in eight specimens. The cell block method revealed negative for malignancy in 52 specimens and positive for malignancy in 13 specimens. The cell block method had a statistically significant higher rate of malignancy detection (20.0% versus 12.3%, p<0.001). Conclusion: The cell block method increased the malignant cell detection rate. In addition, the cell block method has more advantages and the ability to perform further immunohistochemistry staining for a definitive outcome in suspicious cases. [ABSTRACT FROM AUTHOR]

Fine needle aspiration (FNA) cytology is a basic diagnostic technique used to investigate superficial and deep swellings. Rapid on-site evaluation (ROSE) using toluidine blue (TB) is easily available, cheap, cost-effective, can be used both for testing adequacy and giving provisional diagnosis. To evaluate the role of ROSE using toluidine blue staining in arriving at a diagnosis in comparison to routine stains. A total of 1500 cases of FNA of palpable swellings from sites like salivary gland, breast, thyroid, lymph node, and soft tissue lesions [non-image guided] during a 9-month period were studied. All the cases were evaluated by ROSE using toluidine blue stain and routine Giemsa/PAP staining. The results were compared in each case. Only 2% cases proved inadequate on TB, Giemsa and PAP combination, commonest site of inadequacy being lymph node. Adequate sample was obtained within two passes in 92.5% cases. The turn-around time (TAT) was 1 day in 96.4% of cases. The average time for making a provisional diagnosis on TB was 3 minutes. There was 99.2% concordance between TB and final cyto-diagnosis. Validity parameters: sensitivity 98%, specificity 100%, positive predictive value 100%, negative predictive value 99.8%, efficacy 99.2% and false negative 1.94%. ROSE using toluidine blue is a reliable means of demonstrating sample adequacy, for making a provisional diagnosis and guiding collection of diagnostic material for microbiology, immunocytochemistry (ICC), cell block and molecular testing etc. The technique is easy enough for general laboratories to incorporate into their routine practice. ROSE can be called as the "frozen section of cytology". [ABSTRACT FROM AUTHOR]

Background: Cell blocks are being used more frequently in cytology for ancillary testing, including molecular diagnostics. There are several different methods of processing cell blocks, with plasma‐thrombin being one of the most common. Plasma is a blood‐derived product and may be a source of DNA. The aim of this study was to determine whether the plasma used for the plasma‐thrombin cell block method has amplifiable DNA that may potentially interfere with molecular testing results. Methods: Expired bags of fresh frozen plasma were collected from a blood bank. From each sample, DNA was extracted from a 1‐mL aliquot with the QIAsymphony MIDI kit (Qiagen). The concentration of DNA was measured on a NanoDrop instrument. The amplifiable DNA quality was assessed by polymerase chain reaction (PCR) with primers to generate amplicons of various sizes. Characterization was performed with the AmpFLSTR Identifiler Plus PCR kit with capillary electrophoresis. Results: Twenty samples from 20 bags were collected. All samples showed amplifiable DNA despite low DNA concentrations in a few cases. PCR amplification revealed the presence of high‐quality amplifiable DNA (up to 600 base pairs). DNA was amplified at the 16 loci interrogated in all samples tested with the AmpFLSTR Identifiler Plus PCR kit. Conclusions: The presence of genomic DNA in plasma may theoretically interfere with results of molecular testing. Particularly in clinical samples with low cellularity, the DNA in plasma may potentially either mask the presence of minute amounts of tumor‐derived DNA or lead to a false‐positive result. The commonly used plasma‐thrombin cell block method may be a source of DNA contamination. This can potentially interfere with molecular testing as molecular techniques advance and become more sensitive. [ABSTRACT FROM AUTHOR]

Background: The potentially curative and/or palliative therapy for non‐resectable lung cancer has evolved significantly over the past 2 decades. With the availability of targeted therapies, the need for precise sub‐typing of non‐small cell lung carcinoma (NCSLC) has become paramount. Objectives: As there are few data from South Africa, we aimed to determine utility of TTF‐1, napsin A, p63 and CK5 immunostaining on fine needle aspiration (FNA) cell block and formalin‐fixed paraffin‐embedded tissue biopsy specimens in subtyping NSCLC as adenocarcinoma and squamous cell carcinomas. Methods: All cases of NSCLC diagnosed during a 3‐year period were retrospectively identified. All FNA biopsy and formalin‐fixed paraffin‐embedded cases that were stained with TTF‐1, napsin A, CK5 and p63 were collected. A lung cancer registry was used to access and correlate clinical and radiological data. Results: We included 271 cases with diagnoses of adenocarcinoma of the lung (n = 201), squamous cell carcinoma of the lung (n = 53), unspecified NSCLC (n = 8) and other carcinomas (n = 9). TTF‐1 and napsin A had sensitivities of 99.0% and 91.9%, respectively, positive predictive values (PPVs) of 90.8% and 90.3%, respectively, and accuracies of 91.0% for adenocarcinoma of the lung. Napsin A had a higher specificity than TTF‐1 (90.2% vs 62.8%). Both CK5 and P63 had high sensitivities (95.4% and 97.9%, respectively) and negative predictive values of 96.4% and 96.8%, respectively, for squamous cell carcinoma of the lung. CK5 had a higher specificity than p63 (84.4% and 61.2%, respectively), PPV (80.4% and 70.8%, respectively) and accuracy (88.8% and 79.2%, respectively) for squamous cell carcinoma. Conclusion: All four immunostaining methods had high sensitivities. TTF‐1 and napsin A both had high PPV and diagnostic accuracy for adenocarcinoma of the lung, whereas CK5 had an equally high PPV and accuracy for squamous cell carcinoma of the lung. The specificity of napsin A for adenocarcinoma was higher than that of TTF‐1. The specificity of CK5 for squamous cell carcinoma was higher than p63. Utility of TTF‐1, NapsinA, Ck5 and p63 in distinguishing between Adenocacinoma and Squamous cell carcinoma, using fine needle aspiration cellblock material and paraffin embedded biopsy specimens in a resource limited setting. [ABSTRACT FROM AUTHOR]

Besides the established uses in tumor diagnosis and characterization, immunohistochemical assays of effusion fluids are being increasingly used as substrates for prognostic and predictive markers. This commentary highlights validation issues encountered in daily practice that may have an impact on assay interpretation. [ABSTRACT FROM AUTHOR]

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PD-L1 expression assessed by immunohistochemical staining is used for the selection of immunotherapy in non-small cell lung cancer (NSCLC). Appropriate validation of PD-L1 expression in cytology specimens is important as cytology is often the only diagnostic material in NSCLC. In a previous study comprising two different cohorts of paired biopsies and cytological specimens, we found a fairly good cyto-histological correlation of PD-L1 expression in one, whereas only a moderate correlation was found in the other cohort. Therefore, that cohort with additional new cases was now further investigated for the impact of preanalytical factors on PD-L1 concordance in paired biopsies and cytological specimens. A total of 100 formalin-fixed paraffin-embedded cell blocks from 19 pleural effusions (PE), 17 bronchial brushes (BB), and 64 bronchoalveolar lavage (BAL) and concurrent matched biopsies from 80 bronchial biopsies and 20 transthoracic core biopsies from NSCLC patients were stained using the PD-L1 28-8 assay. Using the cutoffs ≥1%, ≥5%, ≥10%, and ≥50% positive tumour cells, the overall agreement between histology and cytology was 77–85% (κ 0.51–0.70) depending on the applied cutoff value. The concordance was better for BALs (κ 0.53–0.81) and BBs (κ 0.55–0.85) than for PEs (κ −0.16–0.48), while no difference was seen for different types of biopsies or histological tumour type. A high number of tumour cells (>500) in biopsies was associated with better concordance at the ≥50% cutoff. In conclusion, the study results suggest that PEs may be less suitable for evaluation of PD-L1 due to limited cyto-histological concordance, while a high amount of tumour cells in biopsies may be favourable when regarding cyto-histological PD-L1 concordance.

Circulating tumor cells (CTCs) are extremely low-frequency cells in the bloodstream. As those cells have detached from the primary tumor tissues and it circulates throughout the whole body, they are considered as promising diagnostic biomarkers for clinical application. However, the analysis of CTC is often restricted due to their rarity and heterogeneity, as well as their short-term presence. Here we proposed formalin-fixed, paraffin-embedded (FFPE) CTC block method, in combination manner with the hydrogel core-mediated CTC accumulation and conventional paraffin tissue block preparation. The hydrogel core specifically captures and releases cancer cells with high efficiency with an immunoaffinity manner. An additional shell structure protects the isolated cancer cells during the FFPE CTC block preparation process. The fabricated FFPE CTC block was sectioned and cytopathologically investigated just the same way as the conventional tissue block. Our results demonstrate that rare cells such as CTCs can also be prepared for FFPE cell blocks and shows great promise for cytopathological CTC studies.

(1) Background: The Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) was introduced in 2018, bringing an organ-specific classification system for salivary gland cytopathology. The aim of present study is to evaluate the MSRSGC prospectively, based on a two-year experience in the tertiary care center pathology department, and evaluate the role of routine cell block (CB) preparation in salivary gland cytopathological diagnostics. (2) Methods: In our institution, the Department of Pathology, Fimlab Laboratories, Tampere, Finland, the MSRSGC has been implemented in salivary gland cytopathology since January 2018 and, over a two-year period (January 2018–December 2019), there were 365 fine-needle aspirations, of which 164 had a surgical follow-up. The CB methods used were Plasma-thrombin, the collection of visible fragments, and the Shandon and in-house methods. (3) Results: The MSRSGC diagnostic figures were as follows: accuracy 87.5%, sensitivity 45.8% and specificity 98.9%. For diagnostic categories of MSRSGC (non-neoplastic, benign neoplasm and malignant neoplasm) (n = 63) diagnostic accuracy was 98.4%, and for undetermined categories (atypia of undetermined significance, salivary gland neoplasm of uncertain malignant potential and suspicious for malignancy) (n = 49) diagnostic accuracy was 73.5%. Non-contributory cell blocks resulted more often in a false negative diagnosis (25%, 3/12) than a true negative diagnosis (10%, 7/73, p < 0.001), and is, most likely, an insufficient cytological diagnosis (86%, 18/21, p < 0.001). (4) Conclusion: The application of MSRSGC and CBs are beneficial in salivary gland cytological diagnosis, increasing diagnostic accuracy and, thus, patients’ management and treatment.

Malignant pleural effusion (MPE) can accompany advanced lung adenocarcinoma. Recent studies suggest that MPE could contain a heterogeneous subpopulation of cells with stem-like properties, such as tumorigenicity and self-renewal, indicating that they could be the source of metastasis. Although previous studies analyzed the correlation between cancer stem cell (CSC) marker expression and clinical outcomes using lung cancer tissues, investigations regarding the association of MPE with CSC marker expression are limited. We performed immunohistochemistry to examine the expression of aldehyde dehydrogenase 1 (ALDH1) and Sal-like 4 (SALL4) in 46 cell block samples of MPE from patients with lung adenocarcinoma. ALDH1-positive and SALL4-positive cancer cells in MPE were detected in 30 (65.2%) and 21 samples (45.7%), respectively. Cluster formation was detected in 26 samples (56.5%). The number of clusters was significantly higher in ALDH1-positive/SALL4-negative samples. SALL4 expression was inversely correlated with the cluster ratio (r = −0.356) and positively associated with the Ki-67 index (r = 0.326), suggesting that MPE cells with high SALL4 expression comprised the proliferative subpopulation. In conclusion, we demonstrated that MPE contains an ALDH1-positive/SALL4-negative subpopulation exhibiting cluster formation and a SALL4-positive proliferative subpopulation.

Background: Breast cancer is a worldwide problem, and early positive diagnosis is critical for establishing the optimal therapeutic strategy. Following a preliminary diagnosis, fine-needle aspirate cytology (FNAC) may be used to obtain cells for immunohistochemical (IHC) analysis and histopathological examination. This study aimed to assess the FNAC method combined with embedding samples in paraffin blocks (cell blocks) and comparing this with core biopsies (tissue blocks). Methods: This observational, prospective study was performed at our hospital and involved 50 female participants who presented with breast masses and were subsequently evaluated for high-risk status by FNAC and IHC. Tests for estrogen receptor (ER), progesterone receptor (PR), and human EGF receptor 2 (HER2/neu) were performed and the sensitivity, specificity, and discrepancy rates between methodologies were calculated using correlation analysis and agreement tests. Results: The correlation analysis between immuno-staining of sections from cell blocks and histopathological examination of sections from tumor-tissue blocks revealed a high concordance for HR and HER2/neu. Conclusion: IHC of cell-block sections was found to be better for the determination of HR status and HER2/neu levels. It is very important to obtain high-quality cell blocks with strict quality control for their clarification.

Background and Aim: Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is a procedure of choice for the diagnostic evaluation of submucosal and periluminal lesions. Tissue sample can be obtained by EUS-FNA cytology (FNAC) or cell block (CB). The aim of the present study is to compare diagnostic yield of EUS-FNA CB and cytology in the absence of onsite pathologist following a protocol-based EUS-FNA approach in solid lesions. Patients and Methods: Participants who underwent EUS-FNA at our center for solid submucosal or periluminal lesions (pancreas, lymph node, and liver) between 2014 and 2016 were included, retrospectively. The indication for the procedure along with the clinical and other investigation details and the final etiological diagnosis were recorded on uniform structured data forms. The diagnostic yield of cytology and CB were compared using McNemar’s test. The P < 0.05 was considered statistically significant. Results: EUS-FNA for solid lesion was performed in 130 lesions in 101 patients during the study period. Their mean age was 52.5 ± 12 years and 42.5% were female. Pancreatic masses were the most common lesions (37.7%) followed by lymph nodes (36.9%). Submucosal lesions (17.7%) and liver lesions (7.7%) accounted for rest of the cases. The overall diagnostic yield for EUS-FNAC (70%) and CB (74.6%) was not significantly different (P = 0.3) and their combined yield was 85.3%. For the 23 patients with submucosal lesion, diagnostic yield of CB (82.6%) was significantly better than cytology (47.8%, P = 0.04). Conclusions: EUS-guided CB has better yield compared to cytology in gastrointestinal submucosal lesions. The combination of CB with cytology improves the overall yield of the procedure; and hence, they should be considered complimentary rather than alternatives.

Aims: Endobronchial ultrasound (EBUS) is a relatively new modality that can be used to guide transbronchial needle aspiration (TBNA) of mediastinal lymph nodes. At present, researches on the sensitivity and specificity of cytopathology based on the EBUS-TBNA are deficient; therefore, we want to evaluate the value of cytology based on the EBUS-TBNA in this article. Materials and Methods: We reviewed the 379 cases that underwent the EBUS-TBNA in Shanghai Pulmonary Hospital from April 2010 to May 2011. Discarding the 139 cases with insufficient cells, we analyzed the remaining 240 cases that had enough cells on the smears. Statistical Analysis Used: The Statistical Package for the Social Sciences version 15.0 (SPSS Inc., Chicago, IL) was used for data analysis. A P value of

Context: Fine needle aspiration cytology (FNAC) of superficial or deep-seated lesion is an increasingly common practice, eliminating time consuming and costly diagnostic procedures and providing rapid and safe diagnosis. Aims: To assess utility of cell block preparation method in increasing sensitivity of cytodiagnosis in deep-seated image-guided FNACs. Settings and Design: This was a hospital-based observational study conducted in the Department of Pathology, over a period of one and a half years. Materials and Methods: A total of 46 cases of abdomino-pelvic and intrathoracic masses subjected to guided FNACs were included. Along with conventional smears, cell blocks were prepared by using AAF (95% Ethanol 34 ml + formalin 4 ml + Glacial acetic acid 2 ml) as fixative agent. Statistical Analysis Used: Done using MedCalc Version 12.7.5.0 to find out the sensitivity, specificity, and diagnostic accuracy of conventional smears and cell blocks. Results: The sensitivity, specificity, and diagnostic accuracy of cell blocks in our study was 71.11%, 100%, and 71.73%, respectively. The figures for FNA smears were 62.22%, 100%, and 63.04%, respectively. Conclusions: Cell block technique by AAF fixative is a simple, inexpensive procedure. Cell block method allows the recovery and processing of minute amounts of cellular material, facilitating better classification of tumor when reviewed along with cytological smears, the ability to obtain many sections for immunostains and other studies to be performed akin to paraffin sections produced in histopathology.

Background/aims: The sensitivities of endoscopic trans -papillary pathologic diagnosis of biliary tract cancer (BTC) are unsatisfactory. Recently, the diagnostic utility of the endoscopic scraper device, Trefle for biliary stricture has been reported. The Trefle can be guided to the target biliary stricture over the guidewire and is as easy to use as the conventional brush catheter (CBC). This study evaluated the efficacy and safety of Trefle-assisted tissue acquisition combined cell block method and CBC cytology for biliary strictures due to BTCs., Methods: We retrospectively reviewed consecutive patients with biliary strictures in whom CBC cytology or Trefle-assisted tissue acquisition under endoscopic retrograde cholangiopancreatography was performed for suspected BTCs from January 2015 to June 2022 at our institution., Results: 173 patients (CBC group; n = 55, Trefle group; n = 118) were enrolled in this study. The sensitivity, specificity, and accuracy of CBC cytology for BTC were 68.3%/100%/76.4%. On the other hand, the sensitivity, specificity, and accuracy of Trefle-assisted tissue acquisition for BTC were 93.7%/95.7%/94.1%, showing superior sensitivity ( p < 0.001) and accuracy ( p = 0.002) compared to that of CBC., Conclusions: Compared to CBC cytology, Trefle-assisted tissue acquisition has superior diagnostic performance while maintaining procedural simplicity and is considered useful for diagnosing malignant biliary stricture., Competing Interests: None., (© 2024 The Authors. DEN Open published by John Wiley & Sons Australia, Ltd on behalf of Japan Gastroenterological Endoscopy Society.)

Background: Claudin-4, a tight junction associated protein expressed in epithelial cells, is purported as a highly specific and sensitive marker for epithelial malignancies. Our aim was to assess the sensitivity, specificity and real-time utility of claudin-4 immunocytochemistry (ICC) in the diagnostic work-up of suspected malignant effusions., Methods: Claudin-4 (3E2C1 clone) ICC was performed prospectively in effusion cell blocks where other ICC markers were being performed as part of reporting over 3 months. Based on claudin-4 staining in unequivocal malignant and reactive effusions, the sensitivity and specificity was calculated. In cases signed out as inconclusive encompassing atypia of undetermined significance (AUS) and suspicious for malignancy (SFM) and negative for malignancy, change in diagnostic category based on addition of claudin-4 ICC was assessed., Results: Study included 107 effusions. Claudin-4 stained 100% of metastatic adenocarcinomas including those with primaries in lung, breast, ovary, female genital tract, gastrointestinal tract and pancreatic-biliary tract, and was negative in all reactive mesothelial and mesothelioma effusions with sensitivity of 100% (48/48) and specificity of 95% (20/21) for adenocarcinoma. Claudin-4 upgraded the diagnostic category to positive in 70% (16/23) of SFM, 20% (1/5) of AUS, and in 50% (5/10) of negative effusions. Among cases with confirmed serosal involvement status on follow-up, claudin-4 showed sensitivity, specificity, positive predictive value and negative predictive values of 85% (11/13), 100% (3/3), 100% (10/10) and 75% (3/4), respectively, for metastatic adenocarcinoma., Conclusion: Claudin-4 as a single marker is sensitive and specific for adenocarcinoma and is a valuable addition to the ICC armamentarium., (© 2023 Wiley Periodicals LLC.)

Cell blocks may be hard to be totally automatically detected by the scanner (ADS), generating incomplete whole slide images (WSIs), with areas that are not scanned, leading to possible false negative diagnosis. The aim of this study is to test if inking the cell blocks helps increasing ADS. Test 1: 15 cell blocks were sectioned, one half inked black (1HB) and the other inked green (1HG). Each of the halves was individually processed to generate a WSI stained by the H&E. 1HBs and 1HGs had similar scanning time (median 59 s vs. 65 s, p = .126) and file sizes (median 382 Mb vs. 381 Mb, p = .567). The black ink interfered less in the observation (2.2% vs. 44.4%; p < .001) than in the green one. Test 2: 15 cell blocks were sectioned, one half inked black (2HB) and the other left unstained/null (2HN). Each of the halves was individually processed to generate three WSIs-one HE, one periodic-acid Schiff (PAS), and one immunostained by cytokeratin AE1&AE3 (CKAE1AE3). HE and PAS WSIs from both 2HN and 2HB groups were all totally ADS and had similar scanning times and file sizes. Concerning immunostaining with CKAE1AE3: ADS (46.7% vs. 93.3%; p = .014), median time for scanning (57 s vs. 83 s; p < .001) and file size (178 Mb vs. 338 Mb; p < .001) were reduced significantly in the 2HN group in comparison with the 2HB. Although increasing scanning time and file size, inking the cell blocks helps increasing ADS after immunostaining, improving the safety and efficiency of the digital pathology workflow., (© 2023 Wiley Periodicals LLC.)

Pilomatrixoma is a relatively rare benign skin appendageal tumor, often presenting in the pediatric age group as a nodular lesion and most commonly involving the head and neck, making it amenable to primary fine needle aspiration (FNA) diagnosis. We report the clinical and histopathological findings of two cases of pilomatrixoma in children, both of which were initially misdiagnosed as small round blue cell tumors due to high cellularity and misinterpretation of the proliferating basaloid cells. Histopathology revealed basal cell proliferation and mitoses indicating that they were progressive, early lesions. The first case showed membranous positivity for CD99 which prompted a diagnosis of Ewing sarcoma. Awareness of the morphological spectrum including positivity for CD99 and careful evaluation of cell block histology could have averted the misdiagnosis. Pilomatrixoma should be included as an important differential diagnosis when faced with primitive-appearing cells on FNA, especially in children with mass lesions in the head and neck region., (© 2023 Wiley Periodicals LLC.)