Cytological Diagnosis of Infectious Diseases: Identification of Pathogens and Recognition of Cellular Reactions

Cytological diagnosis of infectious diseases is as important as the cytodiagnosis of malignancies, because the detection of pathogens in cytological specimens is crucially valuable for prompt and appropriate patients’ treatment. When compared with histological diagnosis, cytology is strong at detecting microbes under Papanicolaou and Giemsa stains. Host response against the infectious agent can be estimated by the type of background inflammatory cells. Patterns of the inflammatory cellular responses against extracellular and intracellular pathogens should be recognized. Immunocytochemical and molecular approaches can be applied, even when we have only one cytology specimen in hand. The cell transfer technique is useful to create plural material from one glass slide for immunocytochemistry and other techniques. In case of transmissible disorders including sexually transmitted diseases, the prompt and appropriate diagnosis will avoid avoidable transmission of infectious agents among people, and eventually contribute to the safety of the human society.

Pancreatic/Peripancreatic Spindle Cell Lesions Cytology: A 15-Year Review

Introduction/Objective Spindle cell lesions (SCL) are diagnostically challenging, especially on cytology specimens where entities have overlapping features. Neoplastic and non-neoplastic SCL are uncommonly encountered in the pancreas/peripancreatic tissue. The sensitivity and specificity of EUS-FNA of pancreatic lesions both approach 95%. In this study, we assess the frequency of SCL found in pancreas/peripancreatic tissue on cytology specimens, the frequency of performing IHC stains and the most useful IHC panels. We also evaluate histology-cytology discrepancies and pitfalls. Methods A retrospective analysis of all pancreas/peripancreatic cytology specimen results between January 2004 - August 2019 was conducted. A total of 5,132 cases were identified. The number of spindle cell lesions was 27 (0.52%), with surgical pathology results available for 15 cases (55%). IHC stains were performed on 18 cell blocks (66%) and 9 surgical pathology specimens (60%). Results Of the 27 SCL identified, 22 lesions were neoplastic (81%), and 5 lesions were non-neoplastic (19%). The Neoplastic cases were: 10 GISTs (37%), 4 spindle cell lesions, NOS (14.8%), 2 metastatic sarcomas (7.4%), 1 pheochromocytoma (3.7%), 1 leiomyoma (3.7%), 1 schwannoma (3.7%), 1 malignant fibrous histiocytoma (3.7%), 1 granular cell tumor (3.7%), and 1 neuroendocrine carcinoma (3.7%). Of the neoplastic cases, 10 were lesions originating in a different organ with direct extension/distant metastasis to the pancreas (71%), with the most common organ being the stomach, 6 cases (60%). The non-neoplastic cases were: 4 granulomas (14.8%), and 1 accessory spleen (3.7%). IHC stains were performed on 18 cell blocks (66%), and attempted unsuccessfully on 2 cell blocks. The most commonly utilized stains were: CD117, 15 cases (83%), SMA, 11 cases (61%), S-100, 9 cases (50%), CD34, 5 cases (27%) and Cytokeratin, 5 cases (27%). IHC stains assisted with proper classification in 9 cytology cases (50%). There was one interpretation error*, and the histology-cytology discrepancy rate was 6%. Conclusion Pancreatic and peripancreatic spindle cell lesions are uncommon (520/100,000), and are particularly challenging on cytology specimens. If neoplastic, the majority originate in an organ other than the pancreas. If non- neoplastic, the majority are granulomas. Immunohistochemical staining is required for proper classification. And in this small series, cytology is highly accurate for diagnosis (96%).

Analysis of Cell Block and Cytology Specimen Preservation from Lung Aspiration Biopsy

<p>Cytology smear technique is often used in Indonesia because the process is safe, simple, easy, fast, and cost effective. At present, several studies have found that smears with cell block techniques are of better quality than smears with cytology smear techniques. The aim of this study was to analyze whether the cytology smear technique can produce adequate specimens compared to cell block towards results of lung Fine Needle Aspiration Biopsy (FNAB). Lung FNAB specimens were divided into two parts: one part was processed with cytology and the other part with cell block technique. The specimens were observed under a microscope to count the number of inflammatory cells and the number of artifacts. The numbers of inflammatory cells and artifacts were scored 0-3. The inflammatory cells consisted of neutrophils, lymphocytes and plasma cells, also macrophages. The result showed no significant difference between the number of inflammatory cells in cytology and cell block (p neutrophils=0.543; p lymphocytes and plasma cells=0.192; p macrophages=0.487) in 38 samples. The artifact score comparison test result showed a significant difference between the number of artifacts in cytology and cell block (p=0.027) with more artifacts in cytology. The most common artifact in cytology was air bubble artifacts, while cell block was dominated by torn pieces artifacts. There was no significant difference between the number of inflammatory cells found in cytology and cell block techniques. Cell block technique has less artifacts than cytology, but artifacts found in cytology can be corrected so that the cytology smear technique is still an option.</p>

Comparison of Fixation Methods for Preservation Cytology Specimens of Cell Block Preparation Using 10% Neutral Buffer Formalin and 96% Alcohol Fixation in E-cadherin and Ki-67 Immunohistochemical Examination

BACKGROUND: Cytological and molecular examinations are among the most important examinations in cancer diagnosis. 96% alcohol is a fixative solution commonly used by clinicians for cytological samples because of its accessibility and affordability. Cellblock preparation from cytology specimen may increase morphology detail and may be used for further biomarker analysis. E-cadherin is an adhesion protein expressed in the cell membrane of most carcinoma. Ki67 is a protein expressed in nuclei of malignant cells that used as a proliferation marker. AIM: This study was designed to investigate the effect of fixation duration in 96% alcohol on protein preservation for immunohistochemistry (IHC) evaluation compared to 10% neutral buffered formalin (NBF) as the gold standard. METHODS: Twenty-five fine-needle aspiration biopsy (FNAB) specimen diagnosed as carcinoma were fixed in 10% NBF and 96% alcohol for 1 hour, 6 hours, 24 hours, 48 hours and 72 hours. Cell blocks preparation were made from those 6 groups of specimens. E-cadherin and Ki67 IHC were done to cell blocks section and evaluated. The data were statistically analysed using the Friedman test with p-value < 0.05 of a significant level. RESULTS: There were significant differences between E-cadherin and Ki67 expression in cell block preparation from 96% alcohol-fixed cytology specimen for 1 hour, 6 hours, 24 hours, 48 hours and 72 hours to 10% NBF (p = 0.0001). CONCLUSION: The result indicated that 96% alcohol is not suitable as a fixative solution for cell block preparation in E-cadherin and Ki-67 IHC examination.

Where in the World Is . . . My Cytology Specimen? Using Lean Six Sigma Methodology in Specimen Management

Introduction An automatic signal was not in place to actively identify when a cytology specimen resulted more than 28 days from collection. The project focus was to decrease the number of specimens resulted more than 28 days and improve result cycle time to enhance safe, quality care delivery. Methods Annually, more than 46% of the cytology specimens processed at our academic medical center originate from our Women’s Health practice site. A Lean Six Sigma Green Belt project commenced with key stakeholders from the practice site, Cytology and Clinical Laboratory Departments. The project goal was to reduce the percentage of specimen results taking longer than 28 days from the baseline of 6.7% to ≤1.7%. Voice of the customer showed 75% of patients would be satisfied with results available within 7 days. Baseline data showed 38.6% of specimens resulted within 7 days with an improvement goal of 85%. Lean Six Sigma methodology was employed to develop standard work. The Cytology Department implemented use of work lists in the CoPathPlus system to track specimens from accession to results. Reconciliation of cytology specimens received versus ordered was performed weekly. Conclusions Baseline data were compared to postprocess implementation data. Hypothesis testing compared baseline to postimplementation cycle time means showing a reduction from 11.28 to 5.15 days (P < .001). Project goal of ≤1.7% for results greater than 28 days was exceeded at 0.22%. Cycle time goal of 85% for results within 7 days was exceeded at 91.6%. Signal presence to identify results greater than 28 days was reduced with improved cycle time for results reporting. With standard work implementation, 616 annualized labor hours and an annualized labor cost reduction of more than $19,000 resulted. The standard work developed has the capability to translate across all practice locations where cytology specimens are obtained.