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Background/Aim: The main objective of microscopic examination of pleural effusions is to ascertain the presence of malignant cells. Effusions prepared routinely using May-Grunwald-Giemsa (MGG)-and Papanicolaou (PAP)-staining can, in a number of cases, provide inconclusive cytological results regarding malignancy. Patients and Methods: This report describes the refined diagnosis of such cases based on immunocytochemical analysis of pleural effusion cell blocks. Of the 340 pleural effusions obtained during 2019 at the Department of Clinical Cytology, Gavle Hospital, Sweden, 63 (18.5%) contained atypical cells of undetermined significance or potentially malignant cells. Results: This diagnosis could be refined using Epithelial Cell Adhesion Molecule/EPCAM (BEREP4) immunocytochemical analysis of effusion cell blocks, allowing previously inconclusive effusions to be classified as clearly benign 42/63 (66.7%) or malignant 21/63 (33.3%). Effusions initially diagnosed as clearly malignant (27/340; 7.9%) were all 27 (100%) BEREP4-immuno-stained. Most BEREP4-positive effusions (37/48; 77.1%) were also carcinoembryonic antigen (CEA) positive. The number of BEREP4-positive cells, however, tended to exceed that of CEA-positive cells. The BEREP4 positive effusions were further examined using different monoclonal antibodies, such as Thyroid transcription factor 1 (TTF-1) for primary pulmonary adenocarcinoma, to determine the original site of the primary tumour. Conclusion: Immunohistochemical staining of pleural effusion cell blocks significantly refines the diagnosis of serous pleural effusions, especially in cases where the preliminary diagnosis was atypical cells of undetermined significance or potentially malignant cells. Furthermore, in the cases of malignancy, the origin of the primary tumour could most often be determined.

Introduction Cytological samples obtained by endobronchial ultrasound (EBUS) are capital for diagnosis, staging and molecular profile in non-small cell lung carcinoma (NSCLC). Objective To assess the success rate of complete, partial and individual of molecular analysis in samples obtained by EBUS-guided transbronchial needle aspiration (TBNA) and/or by oesophageal ultrasound-guided fine needle aspiration with an echobronchoscope (EUS-B-FNA) in patients with NSCLC. Methods Prospective study including 90 patients with non-squamous NSCLC, or non-smoking squamous. Cytological samples were classified into two groups. Group 1: PEN membrane slide and/or cell blocks for the determination of mutations of EGFR, KRAS, ERBB2 and BRAF. Group 2: silane coated slides or cell blocks for rearrangements of ALK, ROS1 and MET amplification. Results The success rate was 78.6% for 4 molecular alterations (EGFR, KRAS, ALK and ROS1), and 44% for 7 determinations. The individual success rate for EGFR was 97%, KRAS 96.3%, ALK 85%, ROS1 82.3%, ERBB2 71.4%, BRAF 67.7% and MET 81.1%. There were no significant differences (p = 0.489) in the number of molecular analyses (1–3 vs. 4) in group 1, depending on the types of samples (cell block vs. PEN membrane slide vs. cell block and PEN membrane slide). Conclusions In patients with NSCLC, the cytological material obtained by ultrasound-guided needle aspiration is sufficient for individual and partial molecular analysis in the vast majority of cases. Membrane slides such as cell blocks are valid samples for molecular analysis.

Aim: Current guidelines recommend evaluating the expression of PD-L1 immunohistochemically in most tumors, especially lung cancer. However, it is not easy to evaluate the immunohistochemical staining of PD-L1 because of the tumor heterogeneity. Therefore, we aim to evaluate the different patterns of PD-L1 in the lung’s nonsmall cell carcinoma and the relationship with clinicopathological features. We also wanted to see if cell blocks obtained from cytology materials can be an alternative for PD-L1 immunohistochemistry.Material and Method: We retrospectively analyzed the immunohistochemical patterns of PD-L1 performed between 2018 and 2019. Biopsy/aspiration procedure of tumors, tumor type, patient’s age, and gender were reviewed. Positive tumor cells (percentage) were categorized according to density and distribution as negative (50%).Results: Material adequacy was found to be satisfactory in evaluating PD-L1 in the cases of cell blocks. Positive staining with PDL1 was detected even with a small number of tumor cells in the cell block. For optimal evaluation of PD-L1 expression in the cell block, the tumor cell ratio of 10% is significant (p=0.002). The cases with negative PD-L1 expression mostly belonged to small biopsy samples (48.3%). However, the difference in PD-L1 expression in histological and cytological samples was insignificant (p=0.79). Besides this, expression of PD-L1 was negative in almost half of the cases (48.3%) diagnosed with squamous cell carcinoma. In adenocarcinoma cases, the PD-L1 expression rate was between 1–49 % in more than half (51.8%) of them. The difference in histological subtype was not significant in PD-L1 staining (p=0.009).Conclusion: In conclusion, we can use cell blocks for immunohistochemical evaluation of PD-L1 expression. Analysis of PD-L1 staining in cytological and histological tissue samples may be a guide for other studies.

BACKGROUND Human papillomavirus-related oropharyngeal squamous cell carcinoma (HPV-OPSCC) presents frequently as metastasis in a neck lymph node that may be cystic or necrotic. Fine-needle aspiration (FNA) biopsies are often first-line diagnostic procedures. p16 immunohistochemistry (IHC) is a surrogate marker for high-risk HPV (hrHPV) infection but can be challenging to interpret. This study evaluated the use of hrHPV in situ hybridization (ISH) in cytology cell blocks of cystic neck lesions. METHODS Twenty-four FNA cases with cell blocks and surgical correlates were evaluated. p16 IHC and hrHPV ISH were assessed on cell blocks (C-p16 and C-hrHPV ISH), and hrHPV ISH on surgical samples (S-hrHPV ISH). All results were classified as negative, positive, or equivocal. RESULTS Two cases were excluded because of insufficient tissue on recut. On the basis of C-hrHPV ISH cases, 12 were positive, 5 were negative, and 5 were equivocal. All 12 positive C-hrHPV ISH cases had concordant S-hrHPV ISH with no false positives. Of the 5 negative C-hrHPV ISH cases, 4 had concordant S-hrHPV ISH, and 1 had a discordant S-hrHPV ISH. Of the 5 equivocal C-hrHPV ISH cases, S-hrHPV ISH were both positive and negative. Fourteen cases were equivocal by C-p16; 9 cases were reliably classified by C-hrHPV ISH (5 positive, 4 negative; 64%). CONCLUSIONS C-hrHPV ISH can be reliably used, especially when positive. A negative or equivocal interpretation of C-hrHPV ISH may warrant repeat testing. Compared to C-p16, C-hrHPV ISH is more frequently diagnostic and could be helpful for HPV-OSCC diagnosis and management.

The detection of the malignant cells by cytologic method has long been practised. The detected malignant cells are difficult to be typed specially in the situations of overlapping cytomorphology. The conventional cytologic preparation may harbour lower number of malignant cells but the method of cell block offers high detection rates of such malignant cells. Still the problem remains when the evaluation is short by morphological features and requires immunoexpression studies. The commonest situation encountered of overlap cytomorphology is the distinction between well differentiated adenocarcinoma from reactive mesothelial cells with atypia in effusion. Such a distinction is not only important for the purposes of diagnosis but also for prognostification by stage and management of the patient .The review presents the experiences of the past studies over utility of EMA in detection and segregation of the cells of well differentiated adenocarcinoma infiltrating the effusions from reactive atypical mesothelial cells. The present review is compiled from the publications from various institute across the globe and shares the authors experience for the utility of epithelial membrane antigen over the cell block of effusions at distinction of overlap cytomorphology. Compilation of the study by inclusion of articles by 1,2,3,3,4,5,6,7,8,9,10.The articles were searched through google engine and Pubmed search. The 10 articles dealing onto the role of EMA as a immunocytochemical marker have affirmed the high sensitivity and specificity of EMA at seggregating glandular malignant cells from atypical mesothelial cells on cell block preparation of body effusions.The range of sensitivity and specificity for EMA was found to be 81% -100% and 98.86 -100% respectively.The positive predictive value range from 92-97% and negative predictive value ranging from 88.64 – 100%. All the studies reviewed confirmed the high value of significance (P value for EMA). The systematic review carried out re-affirms the role of EMA in the situations of of indistinguishable cytomorphology of adenocarcinoma from that of benign , reactive , atypical mesothelial cells. The overall diagnostic utility of EMA was conducted to br over 90% in such situations.

BACKGROUND Fine-needle aspirate (FNA) cytology is often the first-choice method for diagnosing gastrointestinal nodular lesions. The FNA material can be converted to histopathology specimens by a needle rinse cell block (NRCB) technique, allowing ancillary studies to refine the cytologic diagnosis. Despite use in human pathology, NRCB has never been applied to canine or feline gastrointestinal neoplasia. OBJECTIVE This study described NRCB methodology and its diagnostic utility in specific cases of neoplastic gastrointestinal lesions. METHODS Needle rinses with saline were performed after ultrasound-guided FNAs of two intestinal lymphomas (canine and feline) and a canine gastrointestinal stromal tumor (GIST). The NRCB was prepared using the cell tube block technique and processed for paraffin embedding. Routine immunohistochemistry protocols (using CD3, PAX-5, and Ki-67 for lymphoma cases and vimentin, desmin, S-100, and KIT markers for GIST) were applied to NRCB sections, and the results were compared with matched tissue biopsies. RESULTS NRCBs with adequate cell numbers, preservation, and good separation of blood were obtained. The diagnosis and immunophenotyping were confirmed in both cases of lymphoma in NRCBs. In the GIST, the immunolabeling of the neoplastic cells in NRCB was completely concordant with the tissue biopsy. CONCLUSIONS The described methodology is suitable for veterinary settings, having few technical requirements and low invasiveness. The presented cases of gastrointestinal neoplasia highlight the utility of NRCBs as a platform to conduct ancillary studies and refine the cytologic diagnosis.

Oesophageal schwannoma is an extremely rare benign neoplasm. Cytological diagnosis of oesophageal schwannoma has been sporadically reported in the literature. Herein, we present a case of oesophageal schwannoma in an older woman that could be accurately diagnosed based on endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) cytology accompanied by cell block immunocytochemistry.

Serous effusion cytology being a minimally invasive, readily accessible and inexpensive diagnostic procedure. Nevertheless, the accuracy of SEC could vary widely due to the multitude factors including the level of experience of the cytologists Conventional smear has its own limitations with varied efficacy and hence warranting ancillary studies. Cell block has emerged as a paramount and robust platform for sample processing techniques in cytology. Research studies have proved that the efficiency of cytological diagnosis increases by significant margin of 15-20 percent when it is done in conjunction with cell block techniques especially in cases of exudative fluids in picking up crucial cases and based on this, we intended with a novel aim to analyse the accuracy of Serous effusion cytology by combining conventional smear and cell block technique with an attempt to assess the effectiveness of the cell block by our indigenous Modified Bouin’s method. Cell block method prepared by our indigenous Modified Bouin’s preparation with formalin fixative proved to show high quality significance and hence it could be followed in routine practice across laboratories. Cell block technique is quantitatively superior both standalone as well as in conjunction with conventional smear by improving the effective diagnosis of SEC. Diligent use of cell block technique eliminates the suspicious of malignant category on CS and thereby giving more definite diagnosis and thus it is mandated that combined approach of CB in conjunction with CS should be practiced as binary protocol which proved to have obvious influence on patient management. Keywords: Conventional smear, cell block, serous effusion cytology, Histopathology

Introduction: Breast carcinoma is the second most common cancer in the world and establishing an early diagnosis is clinically very important. Fine needle aspiration cytology (FNAC) is the fast, reliable and ideal initial diagnostic modality for the diagnosis of these lesions. Materials and Methods: This is a cross-sectional prospective study of 12 months duration. A total of 160 patients underwent FNAC of palpable breast lump/lumps. A cell-block preparation was done for 80 cases and the histopathological examination has been performed on all the 160 cases. Materials and Methods: The cytomorphological diagnosis, cell-block diagnosis and histopathological diagnosis were correlated using suitable statistical methods. Results: Out of 160 patients, on cytological examination, 107 cases (67%) were benign and 53cases (33%) were malignant. On histopathological examination, 104 cases (65%) were benign and 56 cases (35%) were malignant. On examination of cell block, two cases which were indeterminate in FNAC turned out to be malignant. Conclusion: FNAC plays a main diagnostic role as an out-patient procedure which gives rapid diagnosis. FNAC when combined with cell-block can give more accurate diagnosis. Keywords: FNAC, Breast disease, Cytology, Cell block.

Background: Amyloidosis is menacingly increasing medical problem in aging population across globe. Its clinical presentation is varied. Its laboratory diagnosis often requires biopsy material. The fine needle aspiration cytology (FNAC, FNAB) offers dependable diagnostic alternative to cumbersome biopsy. Present protocol describes the FNAC of abdominal pad of fat as surrogate site for diagnosis of senile systemic amyloidosis avoiding biopsy other known sites. Aim: To study diagnostic accuracy of subcutaneous abdominal fat tissue fine needle aspiration cytology/biopsy for detecting systemic (senile) amyloidosis and its utility in clinical practice. Objectives: To diagnose systemic amyloidosis (senile) by fine needle aspiration cytology/biopsy of abdominal fat pad on crush smears of aspirates and cell blocks. To correlate the diagnosis of deposits of amyloid with the clinical manifestations in specific and non specific clinical manifestations of amyloidosis. To know the sensitivity and specificity of the diagnosis of amyloid deposition on fine needle aspiration cytology/biopsy of abdominal fat pad in comparison to the results of cell block preparations. Methodology: A prospective study carried out to compare results of FNAC of abdominal pad of fat with paraffin embedded cell block in diagnosis of amyloidosis in suspected cases of cases of senile systemic amyloidosis. Technical adopted methods to be used are regular tissue stains, congo red stain and polarized microscopy. Expected Results: It is expected that results of study will establish FNAC of abdominal pad of fat as dependable office diagnostic procedure that would avoid complicated biopsy procedures of gastrointestinal tissue, skin, bone marrow and others for senile systemic amyloidosis.

Objective: To determine the diagnostic yield of cell blocks, prepared by the vapour fixation method, using laboratory supplies easily available at any low resource laboratory setup or outpatient department of a basic healthcare unit. Methods: Prospective descriptive study was carried out at Healthways Laboratories Rawalpindi, for 6 months from 1st January 2017 to 30th June 2017. Walk in patients referred to the lab for FNAC were selected by non probability convenient sampling. After preparing FNAC slides from first pass, the patients were briefed about the cell block material and after informed consent second dedicated passes were done to make cell blocks. Results: Out of 47 cases, there were 25 (53.2 %) breast lumps, 8 (17%) superficial collections, 7 (15.1%) lymphadenopathies, 6 (12.8%) thyroid swellings and 1 salivary gland swelling. Technique yielded moderate to high cellularity in 33 (70.2%) cases, 16 from malignant and 1 from benign breast lumps, 5 from thyroid, 3 from reactive and 3 from metastatic lymphnodes, 3 from abscesses, 1 from lipoma and 1 from salivary gland. Low cellularity 14 (29.8%) cases were from 5 benign and 3 malignant breast lumps, 2 hematomas and 1 case each from thyroid, metastatic lymphnode, abscess and lipoma. Good cellularity was achieved in 16 (84.2%) of 19 cases of malignant breast lesions, 3 (15.8%) were hypo cellular of which 1 was non-diagnostic on FNAC smear. Cell block diagnosis was comparable to FNAC in 35 of 47 (74.4%) cases. Non diagnostic cases on cell block were more, 13 versus 3 cases on FNAC. The kappa value of agreement in diagnosis was 0.64. Conclusions: Poor man’s cell block method is simplest and effective method of cell block preparation which must be advocated at basic healthcare units and low tech laboratory. Its diagnostic potential must be further explored.

Background & Objective: Liver lesions are difficult to diagnose and to differentiate primary from metastatic carcinoma, while Biopsy has its limitations. Cell block technology is easily accessible with high diagnostic accuracy. Our aim is 1) To find the role of cell block technology as an alternative to biopsy in identifying liver lesions; 2) To find the efficacy of cell block along with immunohistochemistry (IHC) and ancillary studies in differentiating primary from metastatic lesions; 3) To identify the site of origin of metastatic lesions. This is a descriptive study undertaken in two tertiary care hospitals over a period of three years. Methods: Retrospective review of adequate samples from fine needle aspirations from liver lesions under radiological coverage, converted into cell block was done. IHC was applied as needed. Usefulness of cell block preparation was evaluated, and the final diagnosis correlated with the biopsy results. Results: Analysis of 323 cases found sensitivity of 98.75% and positive predictive value of 99% for all lesions. Sensitivity for metastatic carcinomas was slightly more than hepatocellular carcinoma. However, accuracy of cell block results for individual metastatic lesions and site of origin was less. IHC and morphological pattern worked as an important adjunct in the final diagnosis. On the other hand, contribution of viral markers as a supplement in the final work up was ambiguous. Conclusion: High precision of validity results of cell block technology in comparison with biopsy highlights its pivotal role in conjunction with supportive tests for diagnosing and differentiating liver lesions as well as identifying primary sites in liver metastasis.

BACKGROUND Fine-needle aspiration cytology (FNAC) is an easy, minimally invasive and useful investigation, and is considered important next to imaging in the rapid diagnosis of lung neoplasm for the last few decades. The purpose of this study was to evaluate the role of image-guided FNAC in pulmonary neoplasm, analyse the results and correlate with cell block histopathological findings. METHODS This is a cross sectional study conducted over a period of two years, involving fifty patients. All the clinical and radiological data were studied from the consenting patient. Then they underwent image-guided FNAC. The cytology smears and cell blocks were evaluated simultaneously to stick out a definitive diagnosis. RESULTS Out of all the 50 cases, we found the necessary FNAC smears and cell block material only in 41 cases. The age range varied from 18 to 90 years with a peak in the fifth to sixth decades. Benign lesions were 4 and malignant were 23 as shown by cytology study. Malignancy was proved in 27 cases in histopathology. The most common tumour was adenocarcinoma (41 %) followed by squamous cell carcinoma (19.5 %). Complications after the procedure were minimal and were noted only in two cases. CONCLUSIONS Image-guided fine needle aspiration cytology of lung tumours provide uncomplicated, cost effective, and a rapid method, for reaching a reliable diagnosis for lung neoplasms with minimum complication. KEYWORDS FNAC, Histological Correlation, Pulmonary Neoplasm

BACKGROUND Cytological examination of intra-abdominal lesions is crucial in diagnosis, staging, and prognosis of suspected malignancies. Fine needle aspiration cytology (FNAC) is a routine diagnostic modality for diagnosis. FNAC is a reliable, rapid, cost effective procedure and plays an important role particularly in low resource centres. FNAC along with cell block has shown remarkable result in diagnosis of malignant lesions and its sub-classification using various antibody markers. METHODS A total of 102 cases of intra-abdominal lesions were studied over a period of one year in this descriptive study. Ultrasound sonography (USG) guided FNACs were done in 89 cases and ascetic / peritoneal fluid analysis in 13 cases. Cell blocks were made in 95 cases and immunohistochemistry (IHCs) were done in 69 cases of suspected / malignant intra-abdominal lesions to confirm the diagnosis. RESULTS Out of 102 cases, cytological smears in 89, fluid smears in 13 and cell blocks in 95 cases were available respectively. Most common intra-abdominal organ involved was liver (21.56 %) followed by retroperitoneum (17.65 %) and lymph nodes (14.70 %). The intra-abdominal lesions were classified into non - neoplastic (16.67 %) and neoplastic lesions (83.33 %). Neoplastic lesions (83.33 %) were further sub classified into benign (15.68 %) and malignant (67.65 %). IHC was done in all malignant lesions for confirmation of primary and / or metastasis. Complete concordance between cell blocks and smears was observed in 95 / 102 cases (93.14 %), while discordances were noted in 05 out of 102 cases (06.86 %). CONCLUSIONS Cell blocks play a vital role in cytopathology and as an adjunct to the routine cytology smears and body fluids. Cell buttons are cost effective and simple to make without involving much higher technicality. Cell block sections offer advantages with respect to cellular architecture, archival storage and application of IHCs which help in pin-pointing the diagnosis. Hence, cell blocks must be considered to increase the diagnostic yield and efficacy. KEYWORDS Cytopathology, Malignant Lesions, Immuno - Histochemistry, Diagnostic Utility

Introduction: The cytological examination of serous effusions helps in diagnosis, staging, management and prognostication of patients with various malignancies. But cytological method has disadvantage of lower sensitivity in differentiating reactive atypical mesothelial cells from malignant cells. Aim: The aim of this study is to compare the cytomorphology of pleural exudative uids by conventional smear (CS) method and cell block (CB) method and also to assess the utility of their combined approach. Furthermore, immunohistochemistry is used in differentiating the benign from malignant tumour cells in exudative pleural effusions. Materials and Methods: It is a cross sectional study. 38 pleural exudative uid samples were subjected to evaluation by both CS and CB methods over a period of 1 year. Cellularity, architecture patterns, morphological features and yield for malignancy were compared. Chi-square and P-value were calculated, p value

Background: The cytological examination of serous effusions has been universally accepted and a positive diagnosis is considered as a definitive diagnosis. It helps in staging, prognosis and management of the patients in malignancies and also helps to differentiate inflammatory and non – inflammatory lesions. The main diagnostic dilemma is to differentiate between reactive mesothelial cells from that of malignant cells in our routine practice. Aims: To compare the morphological features of the conventional cytology with that of the cell block technique. Materials and Methods: The study was conducted in the cytology section of the department of Pathology. Seventy five fluid samples were subjected to diagnostic evaluation for over a period of two years (August 2019 to February 2021). Along with the conventional smears, cell blocks were prepared using 10% alcohol – formalin as a fixative agent. Results: Cellularity and yield for malignancy was more by the cell block technique. Conclusion: The cell block technique provides high cellularity, better architectural patterns, morphological features and an additional yield of malignant cells when compared with that of conventional smear cytology. Keywords: Cell block, Conventional smear, Pleural effusion.

Introduction. Squamous cell carcinoma is a rare form of breast cancer, makes up no more than 2% of invasive breast cancer, usually found in old age. The effectiveness of the cytological research method in the preoperative diagnosis of breast cancer directly depends on the sample preparation of the obtained material. Description of the clinical case. The possibilities of cytological diagnostics are presented using modern methods of sample preparation such as liquid cytology, cell blocks. Conclusion. Features of the cytological picture in combination with clinical and radiological data should become the basis for reliable diagnosis of squamous breast cancer at the preoperative stage.

Introduction Thyroid fine-needle aspiration (tFNA) is a powerful screening tool for assessing solitary thyroid nodules. Generally, morphologic evaluation of smears yields an accurate diagnosis; but, in some cases it is useful to have a cell block (CB) to conduct ancillary studies such as immunohistochemistry (IHC). Cytologic diagnoses guide clinical decisions, so it is important that accurate and efficient diagnoses be rendered. Our study evaluates the diagnostic utility of the CB in the evaluation of tFNAs. Materials and methods We performed a retrospective chart review of all tFNA specimens from January 2014 to July 2019. Data collected included TAT (in days), diagnosis, if a CB was prepared, and if it was diagnostically contributory. Descriptive statistics were calculated. Data were analyzed using the χ2 test and the Mann-Whitney U-test. Results Of the 2321 specimens, 40.2% (933) had CB and only 0.3% (7) were diagnostically contributory. IHC was used for 2 cases. For cases with CB, the median TAT was one day [0-18 days] and the median TAT without CB was 0 [0-9 days]. There was a significant difference in TAT between cases with a CB and those without. Most cases without a CB had same-day TAT (66.4%), whereas only 1.1% of those with a CB had same day TAT. Cases with CB were more likely to have a TAT >1 day (65% versus 12.1%) or >3 days (25.4% versus 10%) than those without a CB (P Conclusions We found the diagnostic utility of CB for tFNAs to be very low. The addition of a CB added at least 1 day to the TAT in all diagnostic strata. The additional time causes patients to wait for results, even for nondiagnostic studies. The increased TAT, resources, and manpower use may be reduced if CB were produced only as needed—if the results of the smear were ambiguous or if ancillary tests were needed to confirm the diagnosis.

Background: Aspiration of serous cavities is a simple and relatively non-invasive technique toachieve diagnosis. Cytological evaluation of body cavity fluid is diagnostically challenging. Especiallyin malignant effusions, helps in staging, prognosis and management of the patients. Aims: Toassess the utility and sensitivity of cell block method over conventional smear technique incytodiagnosis of the serous effusions. And to assess the utility and sensitivity of cytologicalevaluation of body fluids with biochemical and clinical correlation. Methods: A total of 150 fluidspecimens were examined for conventional cytological smear (CS) and cell block method (CB). Outof 150 fluids, 96 were pleural fluid, 48 were ascitic fluid, 04 fluid from pouch of Douglas and 01 wasfrom synovial fluid. Results: In this study, the utility of the CB method in the cytodiagnosis ofmalignant effusions was found to be highly significant as compared to the CS method. The additionalyield of malignancy was 12% more as was obtained by the CB method. Conclusion: For the finalcytodiagnosis of body fluid, there is statistically significant difference between the two techniques. Inother words, CB is superior to CS method. It gives more information about the architecturalarrangement and the likely source of primary. More important is that diagnostic material in cellblocks is available for special studies for Immunohistochemistry which can further supplement ourknowledge about the primary source of metastasis.

Histological evidence is essential for diagnosing malignant biliary strictures. However, conventional brush cytology remains the primary method used worldwide, despite its low diagnostic sensitivity and accuracy, as it is technically easy, rapid, and cost-effective. The aim of this study was to evaluate the diagnostic performance of a recently introduced endoscopic scraper, the simplicity of which is comparable to that of a conventional brush, by comparing diagnostic yields and the number of collected cells. The sensitivity of the endoscopic scraper when using the cell block technique was significantly higher than when using brush cytology or a brush with the cell block technique (53.6% vs. 30.9%, p < 0.001; 53.6% vs. 31.6%, p = 0.024, respectively). Quantitative digital image analysis of cell block sections revealed that the median number of cells obtained with the endoscopic scraper was significantly higher than when using the brush (1917 vs. 1014 cells, p = 0.042). Furthermore, seven cases (8.3%; 7/84) were diagnosed by immunohistochemical analysis of the cell block section obtained from the endoscopic scraper. Given its simplicity and greater capacity for sample acquisition, use of the endoscopic scraper in conjunction with the cell block technique could replace brush cytology for the histological diagnosis of malignant biliary strictures.

Objectives: To determine the role and efficacy of fine needle aspiration cytology (FNAC) and cell block in diagnosis of jaw lesions and compare the agreement between FNAC and cell block to predict the diagnosis. Method: The sample comprised 51 cases, including 12 odontogenic keratocysts (OKCs), 8 ameloblastomas, 22 radicular cysts, 7 dentigerous cysts, and 1 each of intraosseous mucoepidermoid carcinoma (MEC) and adenomatoid odontogenic tumor (AOT). FNAC samples remaining after hematoxylin and eosin (H&E)-stained cytosmear diagnosis were centrifuged at 3,000 rpm for 10 min. The supernatant was discarded and sediment mixed with 2–3 mL alcohol and filtered. To this, 10% formalin was added, filtered, taken for routine processing, and stained with H&E. The result of FNAC smear and cell block was compared with histopathological diagnosis. Results: On cytological examination of the smears, 7 OKCs and 22 radicular cysts were diagnosed, whereas ameloblastomas, AOT, intraosseous MEC, and dentigerous cysts were not. This gave an agreement of 56.8% with the biopsy reports. Cell block sections stained with H&E of 12 OKCs, 22 radicular cysts, 1 MEC, and 3 cases of ameloblastoma offered a diagnosis in accordance with the biopsies giving an agreement of 74.5%, while dentigerous cyst and AOT failed to do so. In comparison with FNAC, additionally 5 cases of OKC and 1 of MEC could be detected, and in ameloblastoma, out of 8 cases, only 3 yielded a concordant diagnosis through the cell block technique. Conclusion: In comparison with FNAC, the architectural pattern and the morphology of the cells were better preserved by the cell block technique. This substantiates that cell block could be used as an ancillary technique to aid in definitive diagnosis of head and neck swellings.

The use of the cell block in the cytology laboratory can be a routine procedure, having an important role in establishing the diagnosis and also in performing additional tests without the need for additional sampling of biological material. Numerous attempts have been made to modify and improve the cell block over the years, however, obtaining a cell block from a fluid biological product is a current challenge because lack of cell adhesion to a tissue remains the most common reason for dissatisfaction among cytopathologists. Although various methods for obtaining cell blocks have been proposed and described in the literature, it is noted that each of them has a disadvantage, in addition to advantages. Thus, in this study we propose a new method that eliminates in turn the disadvantages of other methods, obtaining an increase in cell densities, can apply additional testing applications, while minimizing laboratory costs.

Immunocytochemistry (ICC) is an important ancillary technique in clinical cytology for not only identifying and characterizing tumor cells but also gaining prognostic or therapeutic information. Although cell blocks are often prepared for immunocytochemical evaluation of body cavity fluid and fine-needle aspiration specimens, they are not suitable for hypocellular samples. Liquid-based cytology can help prepare additional smears from residual cytological specimens. However, since conventional methods are used for nongynecological specimens in most laboratories, ICC is often limited by the number of cytological smears. Cell transfer methods permit to evaluate several immunocytochemical markers in a single cytological smear. Yet, these methods have some limitations; for example, they are time-consuming (about 3–40 h) and medium membranes with their attached cells are occasionally stretched or torn when peeled off the slides. Therefore, in an attempt to solve these problems, we developed a rapid and reliable cell transfer method using a nylon mesh. Our method requires no special equipment or reagent and can significantly reduce the turnaround time, as compared to previous methods.

Objective: The cell block (CB) is an important adjunct to cytological preparations in diagnostic cytopathology. Optimizing cellular material in the CB is essential to the success of ancillary studies such as immunohistochemistry (IHC) and molecular studies (MS). Our aim was to identify which CB method was most suitable in a variety of specimen types and levels of cellularity. Study Design: We assessed 4 different CB methods, thrombin clot method (TCM), MD Anderson method (MDAM), gelatin foam method (GFM), and agar method (AM), with descriptive observations and ranking of the methods based on quantity of cells and morphological features. Results: TCM performed best in ranking for both quantity of cells and morphological features, followed by MDAM, GFM, and AM. Lack of adjuvant in the MDAM resulted in some unique morphological advantages which, however, also resulted in inconsistent performance. In low cellularity cases insufficient cells were frequently identified on slides from MDAM and AM CBs. Technique touch time was similar for all methods, with total processing time being shortest for TCM followed by MDAM, GFM, and AM. Conclusions: TCM was the most robust CB technique, retaining high scores for ranking of quantity and morphology in a variety of specimen cellularities and specimen types.

Background Orbital hematolymphoid lesions are rare and usually encountered in elderly patients. Orbital lesions are not easy to biopsy: hence fine needle aspiration cytology (FNAC) can be a very good diagnostic modality for these lesions. Materials and methods Cases of orbital masses subjected to FNAC dating from 2013 to 2020 were retrieved from our archives. A total of 16 cases with biopsy confirmation were included. All clinical details, the type of procedure, details of the immunocytochemistry (ICC) performed on smear, follow-up biopsy, and their haematological work-up were analysed in detail. Results Sixteen biopsy-confirmed cases had been diagnosed as orbital haematolymphoid lesions on cytomorphology and further categorised with ancillary studies including ICC. In twelve instances, the cytology impression was congruent with the histopathological diagnosis and eight of the sixteen cases (50%) proved to be primary orbital lymphoma. Four were secondary orbital lymphomas and the remaining four included one case each of plasmacytoma, myeloid sarcoma, Rosai-Dorfman disease and angiolymphoid hyperplasia with eosinophilia. Conclusion FNAC is a minimally invasive procedure for diagnosing most of the haematolymphoid orbital lesions and it has a rapid turnaround time. The accuracy of cytomorphology combined with ICC on smears/cell blocks can be as good as a biopsy for exact categorisation. Additionally, aspirate smears are preferred samples for cytogenetics compared to formalin-fixed tissue blocks, as molecular cytogenetics techniques are frequently employed for diagnostic, prognostic, and therapeutic purposes.

The WHO 2016 update on lymphoid neoplasms incorporates criteria independent of tissue architecture in the diagnosis and sub-classification of non-Hodgkin lymphoma. Thus, lymphoid neoplasms with characteristic immunophenotypes and/or cytomorphology could be classified with FNA material with appropriate use of ancillary methods. We analysed the efficacy of cytomorphology and cell block immunocytochemistry (CB-ICC) in in diagnostic workup of non-Hodgkin lymphoma diagnosis on FNA material. 65 clinically suspected cases of lymphoma presenting with lymphnode enlargements were analysed. FNAC and CB-ICC were performed and compared with the subsequent biopsy. The sensitivity, specificity, PPV and NPV of CBICC in the diagnosis of NHL were 84.6%, 75%, 95.6%, and 42.8%, respectively in our study. We also observed that use of CB-ICC can have significant logistic implications in patient management in terms of reducing the turn-around time to diagnosis and improving the accuracy in diagnosis on FNAC. ICC on cell blocks can easily be done on FNA material and is a very useful adjunct to FNAC in establishing the diagnosis in a suspected case of NHL especially when a biopsy is not available. Keywords: Cell block immunocytochemistry, NHL, FNAC, Ancillary methods.

Purpose: To further evaluate the value of a standardized method to analyze vitreous samples using the Cellient® automated cell block system in the diagnosis of unsolved uveitis.Methods: Six hundred sixty-four pure vitreous samples obtained from patients with unsolved uveitis between March 2012 and May 2018 at University Hospitals Leuven, fixed in PreservCyt® and processed by the Cellient tissue processor, were included in the study.Results: In 86.3% of the cases, sufficient material was found for diagnosis. A diagnosis of acute inflammation was made in 20.2% of the cases. In 34.0% of the cases, the diagnosis was chronic active inflammation; in 26.5%, low-grade inflammation; and in 5.6%, a malignant process.Conclusion: Our standardized protocol can be used to diagnose endophthalmitis, lymphoma, granulomatous disease, inactive and storage disease, and metastatic infiltration. Cellient analysis of vitreous specimens had a diagnostic yield of 86.3% in unsolved uveitis cases.

Background Paediatric abdominal neoplasms are fairly common. Fine needle aspiration cytology (FNAC) is used for the initial evaluation of paediatric abdominal neoplasms. However, sometimes FNA interpretation can be difficult on limited material, owing to tumour heterogeneity and overlapping features. Therefore, we attempted to obtain additional information using cell block (CB) preparation from residual aspiration material along with immunohistochemistry (IHC) to enhance the diagnostic accuracy. Aim To evaluate the correlation between CB preparation and FNAC in diagnosis of paediatric abdominal tumours with the help of an extended panel of IHC markers and to highlight some of the diagnostic difficulties. Materials and methods A total of 113 cases of paediatric abdominal masses were studied. In addition to routine FNA smears, CBs were prepared from the residual material for IHC analysis as considered appropriate. Results This study included a total of 113 children with abdominal masses. Histopathology following surgical resection was available in 53 cases. Histology was taken as gold standard to measure the diagnostic accuracy with reference to sensitivity (Sn), specificity (Sp), positive predictive value and negative predictive value. The Sn of FNA alone was 87.5% and Sp was 97.78%. The Sn and Sp increased with use of CB alone and were 100% and 97.78% respectively. The highest Sn and Sp were observed when CB was combined with IHC where both the values were 100%. Conclusion CB with IHC is a useful adjunct to the routine FNA smears that further contributes to enhance the accuracy of the cytopathological diagnosis and is useful for choosing pre-operative chemotherapeutic regimen.

Introduction Conventional cell blocks (CCB) prepared from cytological specimens are very useful but the method is relatively time-consuming. Suitable modifications in cell-block techniques are beneficial for improving the turnaround time. We share our experience of a rapid microwave cell-block (MCB) technique. Aim and objectives To study the quality of routine and immunohistochemical (IHC) staining of cell-block sections from serous body fluids prepared by the MCB technique compared with the CCB technique. Method A total of 177 serous body fluid samples were processed by routine centrifugation technique, and the sediments were used for cell-block preparations by both conventional and rapid microwave methods. Cell-block sections were stained with haematoxylin and eosin stain. Haematoxylin and eosin staining quality was analysed using three parameters (cellularity, morphology and staining intensity). IHC for epithelial membrane antigen and calretinin were also performed, and the quality of staining was evaluated on 62/177 samples. Results were analysed using appropriate statistical tests. Results The time taken for processing cell blocks by the MCB method was 1 hour and 18 minutes compared to 13 hours and 45 minutes by CCB. The quality of sections by both methods showed good agreement for cellularity and intensity of staining, and moderate agreement for morphology. A 100% concordance was noted for distinguishing benign and malignant samples on morphology as well as with IHC stain results. Conclusion Although the techniques are comparable in terms of quality of routine and IHC staining, we recommend using the MCB technique due to its short turnaround time.

Objectives Endocervical curettage (ECC) specimens may be limited by scant tissue. We evaluated whether a cellular concentration processing method could improve their diagnostic quality. Methods Between October 2018 and June 2019, ECC specimens were assigned chronologically to one of two groups: nonconcentrated ECC (NECC) or concentrated ECC (CECC). NECC specimens underwent routine histologic processing. CECC specimens were processed using a published HistoGel-based cell block method. We reviewed diagnoses for ECCs, concurrent cervical biopsies and/or loop electrosurgical excision procedures (LEEPs), and preceding Papanicolaou (Pap) smears. We performed multivariate logistic regression analyses to evaluate the impact of processing method on ECC adequacy and discordance between Pap smear and worst tissue diagnoses. Results NECC and CECC adequacy was 88.2% and 84.7% (P = .06). ECC adequacy was greater if concurrent biopsy/LEEP was performed (odds ratio [OR] = 1.76, P < .01). Discordance between Pap smear and worst tissue diagnoses was 9.5% and 13.3% (P = .04) for cases with NECC and CECC processing, although processing method was not significant in multivariate analysis (OR = 0.74, P = .11). Adequate ECC sampling and concurrent biopsy/LEEP were independently associated with concordance between Pap smear and worst tissue diagnosis (OR = 0.46, P < .01 and OR = 0.65, P = .02). Conclusions ECC processing method did not significantly affect either specimen adequacy (P = .06) or diagnostic discordance (P = .11) when controlled for other factors.

Background Although fine needle aspiration cytology (FNAC) is highly accurate for detecting breast malignancies, concerns remain among cytopathologists about false-positive and false-negative diagnoses. Cell block (CB) preparations have been advocated by some cytopathologists as one of the methods to improve and consolidate the diagnostic accuracy of FNAC. The aim of this study was to determine the diagnostic utility of CB in FNAC of palpable breast lesions among female patients. Methods Following FNA, CBs were prepared using 10% neutral-buffered formalin from the residual breast aspirates of 100 consecutive female patients attending the FNAC clinic. The slides of the conventional smears, CB and excisional biopsies were examined, and results were analysed using the SPSS. Results Of the 100 patients that had FNAC, 44 (44%) had excisional biopsy performed. An additional 13% diagnostic yield for malignancy was obtained with the use of CB preparations. CB reduced equivocal diagnoses by 25%, corresponding with 90.9% improvement on definitive diagnoses. Conclusion In our setting, the addition of CB to smear remarkably improved the diagnostic utility of breast FNAC by minimising atypical and suspicious for malignancy diagnostic categories.

Background: The various advantages of Fine-needle aspiration (FNA) biopsy/ cytology can be limited due to inadequacy of the specimen and expertise required for diagnosis. Cell blocks (CBs), though routinely used in cytology of body fluids, can also be used in FNA material with considerable increase in diagnostic accuracy. Aims & objectives: To compared FNA & CB as diagnostic tool and to evaluate whether cell block adds to the diagnostic accuracy of FNA. Methods: This is a two years prospective study carried out in the pathology department of a tertiary heath care hospital and medical college of South India which included the cases undergoing the FNAC, and followed by the biopsy. CB was prepared using Tissue coagulum clot (TCC) method. The smears and tissue sections were assessed for cellularity and adequacy for diagnosis. CBs were grouped into four diagnostic categories. FNA & CB were compared with histopathology (HPE) diagnosis to calculate Sensitivity, Specificity and Diagnostic Accuracy. Result: A total of 195 cases were included. Cellularity and adequacy for diagnosis were higher in CB. They were diagnostically better (superior) in 13.84% (27) cases. The sensitivity (95.77%) and diagnostic accuracy (94.87%) of CB was found to be considerably higher than FNA (78.84% and 78.46% respectively). Conclusion: In cases of suspicious/ intermediate diagnosis or diagnosis discordant with clinical ones, by FNA; should be followed with the CB to improve the diagnostic yield & to facilitate accurate diagnoses. TCC method is a simple CB method which has higher diagnostic accuracy than FNA.

Background: There are not many studies conducted in India to compare cell block preparation methods with reagents and materials that are readily available in all laboratories. This study aimed to standardize and compare two simple cell block techniques, which can be done in low resource settings too. In the study, 35 cases of thyroid, lymph node, and breast were collected for both FNA and cell block preparation for six months. Materials and Methods: There were separate passes given for both methods. A total of seventy cell blocks made using formalin and agar methods of preparation. Results: We compared both the methods on technical and morphological levels. The formalin method was overall easy to perform and was yielding good morphological results in 98% cases, the only drawback being cell loss during handling and processing. While in the agar method, there was almost no cell loss, but it was more technically difficult and yielded poorer morphological results. A scoring system was made for cellularity: no cells = 0, hypo-cellular = 1+, hypo-cellular with tissue fragments = 2+, cellular = 3+.18 A score of 2+ and 3+ was scored by 31/35 formalin blocks and 28/35 agar blocks. Conclusions: The sensitivity of both formalin and agar methods are almost comparable. However, the procedure of the formalin method is far more straightforward and user friendly. Moreover, it also provides a better architectural picture than the agar method.

Aim Primary thyroid lymphoma (PTL) is a rare malignant disease. Its prognosis depends on early diagnosis. The role of fine-needle aspiration (FNA), including smear cytology, cell block (CB) techniques, and immunohistochemistry (IHC) sections in the diagnosis of PTL is still unclear. Here we reported 19 cases of PTL and literature review to evaluate the diagnostic accuracy for lymphoma by cytology. Methods Our study retrospectively reviewed 19 patients diagnosed with PTL at the affiliated hospital of Southwest Medical University in China from June 2011 to May 2019. According to the Bethesda system for reporting thyroid cytopathology, the CB sections were evaluated for the presence of single tumor cells. IHC was performed on CB. Results The diagnostic accuracy for PTL of FNA, CB with smears, and the joint application of the three methods (FNA + CB + IHC) of our study with 19 cases was 68.4% (13/19), 83.3% (15/18), and 100% (17/17), respectively. Conclusion The present study demonstrates that FNA has low sensitivity in diagnosing PTL, but the joint application of FNA, CB, and IHC might provide high diagnostic accuracy for lymphoma and should be applied in all cases where the clinical suspicion is high regardless of the FNA findings.

The role of fine needle aspiration cytology in diagnosing recurrent and metastatic sarcomas has been well dealt with, however; its utility in rendering a primary diagnosis of sarcoma still remains a topic of debate. The present study evaluates the scope of FNAC along with adjunctive techniques including cell blocking and immunocytochemistry in diagnosing and subtyping of soft tissue tumors. A total of 71 patients presented with musculoskeletal lesions underwent fine needle aspiration. Cell blocks were contributory in exact subtyping in 41/56 (73.2%) cases. Round cell tumors were the most easily subtyped tumors with 19/20(95%) subtyped accurately. Similarly, 8/17(47.1%) of pleomorphic tumors, 9/19(47.3%) of spindle cell tumors, 1/1(100%) of myxoid tumors, 0/1(0%) epithelioid tumors, 0/2(0%) of lipomatous tumors, 1/2(50%) of mixed tumors and 3/9(33.3%) of other tumors could be accurately subtyped using ancillary techniques. Histopathological correlation was available in 51 cases.

Background Variability in preanalytical and analytical steps for immunocytochemistry (ICC) on cytology samples is poorly defined. The objective of this study was to evaluate current practices for ICC on cytology samples in European laboratories. Methods A link to an online survey with 19 questions about ICC practices was distributed to cytology laboratories through national representatives in the European Federation of Cytology Societies. Results In total, 245 laboratories responded to the survey by January 30, 2019. Cell blocks, cytospins, liquid-based cytology (LBC) preparations, and smears alone or in combination with other preparations were used for ICC in 38%, 22%, 21%, and 19% of laboratories, respectively. In general, various combinations of preparations were used for ICC in greater than one-half of laboratories (147 of 245; 60%), whereas only 1 specific type of cytology preparation was used in the remaining 98 of 245 laboratories (40%) laboratories. The majority of laboratories (217 of 226; 96%) performed ICC on automated platforms using protocols that were the same as those used for formalin-fixed, paraffin-embedded samples (238 of 527 laboratories; 45%), either optimized (138 of 527 laboratories; 26%) or optimized and validated (151 of 527 laboratories; 29%) for cytology preparations. Positive control slides, negative control slides, and external quality control were used in 174 of 223 (78%), 112 of 223 (50%), and 111 of 120 (50%) laboratories, respectively. Greater than 1000 ICC tests were performed yearly in 34% of laboratories (65 of 191; average, 1477 tests; median, 500 tests). Conclusions ICC is extensively performed in European laboratories using variously prepared cytology preparations on automated platforms, mostly without quality-assurance measures.

Cell blocks and fine needle aspirations can be used for cytopathological diagnosis. Conventional fine needle aspiration smears provide limited material for diagnosis. The cell block technique provides more tissue, which improves diagnostic accuracy. We compared a modified cell block cytology to fine needle aspiration for providing optimal preservation of histochemical and immunocytochemical properties. We used 30 fine needle aspirates from oral lesions in two groups: group 1, fine needle aspiration cytology; group 2, cell block cytology. Smears of fine needle aspirates were stained with Papanicolaou. For the modified cell block technique, aspirated material was centrifuged to create a cell pellet, which then was fixed with Nathan alcohol formalin substitute. After routine histopathological processing, cell pellets were embedded in paraffin, then sectioned and stained with hematoxylin and eosin. Sections were compared to Papanicolaou stained smears of fine needle samples. Cellular morphology and staining quality of modified cell block samples were superior to fine needle aspiration cytology; both methods exhibited distinct nuclear morphology. Modified cell blocks provide excellent cytopathologic features compared to fine needle aspiration cytology.

BACKGROUND Cytology of the adrenal gland is a less performed technique even in tertiary care centres; yet cytological evaluation is an important diagnostic tool for assessment of adrenal lesions. Our objective was to evaluate the diagnostic utility of FNAC smears and cellblock with immunohistochemistry (IHC) in lesions of the adrenal. MATERIAL AND METHODS We had a total of 50 cases over a period of 2 years where both FNAC smears and cellblocks were taken. The smears and cellblocks were examined for adequacy. They were subsequently categorised into four groups: unsatisfactory, benign, suspicious of malignancy and malignant. The diagnostic accuracy of FNAC smears and cellblock with IHC were evaluated and compared, taking histopathology, wherever available, as the gold standard, RESULT: We had 50 cases with age ranging from 6 to 53 years with a median of 7.5 years. Of these, 54% were cytologically malignant and neuroblastoma was the commonest lesion. Histopathology was available in 23 cases only, where the diagnostic accuracy was evaluated. The sensitivity and specificity of FNAC smear was 100% and 85.71%, respectively whereas the sensitivity and specificity of cellblock with IHC was 100% and 92.86%, respectively. CONCLUSION Cellblock together with IHC provides a higher degree of specificity, reduces the unsatisfactory rate and improves the diagnostic accuracy in lesions of the adrenal gland. Immunohistochemistry is an important adjunctive tool.

Introduction In the selection of non-small cell lung cancer (NSCLC) patients for immunotherapy, specimen processed as cell blocks (CBs) may be the only available material to assess PD-L1 expression. Therefore, optimal CB preparation becomes paramount. In this context, here we assessed whether inadequate fixation time might be one of the pre-analytical factors affecting PD-L1 expression. Methods Ex vivo CBs from placental (n = 3) and NSCLC (n = 8) resection specimens were obtained. PD-L1 staining was performed on CBs prepared at increasing fixation times (12 hours, 48 hours, 72 hours, 96 hours, 168 hours and 504 hours) using the companion diagnostic SP263 Assay and a validated 22C3 laboratory developed test (LDT). Staining intensity and percentage of positive cells were evaluated. Results All placental CBs showed moderate to strong PD-L1 positivity in most cells, regardless of the fixation time. Likewise, the percentage of SP263-stained NSCLC cells was similar at all fixation times except for one case, which showed less intense SP263 staining at 168 hours. Conversely, in 5/8 cases, the 22C3 LDT percentage of positive cells and staining intensity decreased at 168 hours and 504 hours. Conclusions Our results show that fixation time influences the performance of 22C3 LDT on CBs. Thus, we recommend that the fixation time of cytological materials be carefully checked, especially when PD-L1 testing is delayed until the oncology request. Indeed, delays in tissue processing and paraffin embedding may lead to sub-optimal performance of PD-L1 staining on CBs.

Introduction Immunotherapy has shown promising results in non-small cell lung cancer (NSCLC), for which tumour-infiltrating cytotoxic (CD8+) T cells play a critical role. We investigated the utility of image analysis (IA) to quantify CD8+ T cells in a series of matched small biopsies and resections of NSCLC. Methods CD8 immunohistochemistry was performed on cell-blocks (CB), core needle biopsies (CNB) and corresponding resections from primary NSCLCs. Slides were digitised using an Aperio AT2 scanner (Leica) and annotated by whole slide image (WSI) or fields of view occupied by tissue spots (TS). Quantitative IA was performed with a customised Aperio algorithm (Leica). CD8 scores (number of T cells with 1-3+ staining/total area) were then compared. Results Forty-four cases with CB or CNB material and a corresponding resection were analysed. Average CD8 score was determined in CB (7.67 WSI, 77.67 TS) and/or CNB (47.35 WSI, 325.67 TS), and corresponding resections (190.35 WSI, 336.58 TS). CD8 score concordance was highest (78.6%) for CNBs using WSI annotation. Overall, small biopsies (CB or CNB) correlated with the resection in 71.4% cases using WSI and 63.3% cases using TS annotation. IA performed better for low CD8 scores. Conclusions These findings show that CD8 density in NSCLC can be quantified by IA in small biopsies and cell blocks, achieving the best concordance using WSI scores. Discrepancies were attributed to values near the cut-off and background detection of staining. These data warrant future studies with more cases and follow-up data to further investigate the clinical utility of IA for CD8 analysis in NSCLC.