Diagnostic utility of cytospin, cell block and immunocytochemistry in pleural effusion cytology

Background: With the introduction of cytospin, the sensitivity of diagnosing malignancies has increased mainly due to the increase in cellular yield. Cell block also gives the advantage of ancillary testing and allows for retrospective studies. Immunocytochemical markers are used to differentiate and subtype various malignancies in body effusions.Aim of the study was to compare the morphological features of both technique and to assess the diagnostic utility of cell block methods in the cytodiagnosis of pleural effusions.Methods: This was a Prospective observational comparative study of two cytopreparatory techniques. All samples were examined and processed by cytospin and cell block techniques. Continuous data were expressed as Mean±SD (standard deviation) while categorical data were expressed in number, percentage and compared by chi-square (χ2) test.Results: The final diagnosis of both cytospin (147 cases) and cell block (150 cases) techniques was divided into four broad categories: Inadequate, Benign, Suspicious and Malignant. The significant diagnostic cytospin (AUC=0.857, p<0.001) in discriminating positive and negative malignant cases with 75.00% sensitivity (95% CI=53.3-90.2) and 100.00% specificity (95% CI=86.7-100.0) and with 100.0% positive predictive value and 81.2% negative predictive value. In contrast, cell block also showed significant diagnostic but with higher accuracy (AUC=1.000, p<0.001) and sensitivity 100.00% (95% CI=86.7-100.0) and specificity 100.00% (95% CI=86.7-100.0) and 100.0% positive predictive value and 100.0% negative predictive value than cytospin technique.Conclusions: Cell block as a technique should be used in routine practice as it not only increases the diagnostic yield but ancillary test can also be done.

Hallazgos del lavado broncoalveolar procesado con técnica de cytospin y citología convencional / Findings in bronchoalveolar lavage processed using a cytospin technique and convential cytology

Actualmente la interpretación de la citología en lavado broncoalveolar (BAL) se realiza por medio de técnicas de citología convencional o en cytospin. Esta última obtiene preparaciones concentradas en un área de 6mm para su visualización, con lo que se disminuye el tiempo de lectura por cada muestra, sin deterioro de la calidad.Objetivo: Determinar la diferencia del tiempo de lectura y la calidad de la técnica convencional comparada con la técnica en cytospin.Diseño del estudio: Estudio de corte transversal. Se incluyeron muestras de BAL. Las muestras se procesaron con técnica convencional y con técnica por cytospin. Se comparó el tiempo de lectura para el diagnóstico de cada técnica. Se evaluó la calidad considerando: preservación celular y extendido inflamatorio. La diferencia de tiempos entre las técnicas se determinó por la prueba de Wilcoxon.Resultados: Se procesaron 80 láminas de BAL: 40 casos con la técnica convencional y 40 casos con la técnica en cytospin. A 6 casos no se les realizó conteo diferencial, 4 por exceso de células escamosas y 2 casos en la técnica convencional por no completar 300 células. Se encontró degeneración marcada de las células epiteliales en la técnica convencional con diferencia significativa en el tiempo de lectura con un valor p (0,0000).ConclusionesSe obtuvo diferencia significativa en los tiempos de lectura de cada técnica. Este fue mayor en la técnica convencional que en la técnica con cytospin, con extendidos de mejor calidad en la técnica por cytospin...

The cytological interpretation of bronchoalveolar lavage (BAL) specimen is currently performed using conventional cytology and cytospin preparations. The latter features concentrated preparations in a 6-mm area for sample visualisation and reduced reading time, with no deterioration in quality.Objective: To determine the differences in reading time and quality, between the conventional technique and the cytospin technique.Study designA cross-sectional study was conducted on BAL specimens that were processed using the conventional technique and the cytospin technique. Reading time for diagnosis between the 2 techniques was compared. Quality was assessed taking into account, cell preservation and inflammatory cells in smears. The difference in time was calculated using the Wilcoxon test.Results: Eighty (80) BAL slides were examined, 40 by the conventional technique and 40 by the cytospin technique. A differential count was not performed in 6 cases, in 4 due to excessive number of squamous cells and in 2 cases, processed by the conventional technique, due to exhibiting less than 300 cells. A marked degeneration of epithelial cells was observed using the conventional technique, showing a significant difference in reading time (P<.001).ConclusionsA significant difference was found between reading times of the 2 techniques, which was longer when using the conventional technique. Better quality smears were observed when using the cytospin technique...

Strongyloides stercolaris en lavado broncoalveolar / Strongyloides stercoralis in bronchoalveolar lavage specimen

Las estrongiloidiosis es una infección parasitaria frecuente en zonas tropicales y subtropicales. Suele ser asintomática y limitarse al intestino. Sin embargo, pueden darse casos de infección extraintestinal diseminada y potencialmente fatales en pacientes inmunocomprometidos. Se presenta el caso de una paciente diagnosticada con estrongiloidiosis mediante una muestra de lavado broncoalveolar procesada con los métodos de cytospin y citología convencional...

Strongyloidiasis is a parasitic infection found especially in tropical and subtropical regions. It is usually an asymptomatic and limited disease of the gut. However, potentially fatal cases of disseminated hyperinfection in immunosuppressed patients can occur. We present the case of a female patient with strongyloidiasis in bronchoalveolar lavage specimen processed as cytospin preparations and conventional cytology...

A Comparison Between ThinPrep Monolayer and Cytospin Cytology for the Detection of Bladder Cancer

PURPOSE: The sensitivity of urine cytology is higher for carcinoma in situ and poorly differentiated tumors in bladder cancer, while being fairly low for low-grade or well-differentiated tumors. Development of a sensitive diagnostic test to detect bladder carcinoma would significantly facilitate patient management and allow earlier treatment of this disease. This study compared ThinPrep urine cytology (Cytyc Co.) and conventional Cytospin urine cytology (Shandon Scientific Ltd.) in the diagnosis of bladder cancer. MATERIALS AND METHODS: From January 2002 to December 2010, ThinPrep cytology and conventional urine Cytospin cytologic examination of bladder washings were performed in 3,085 subjects suspected of having bladder cancer and in 379 patients with follow-up after transurethral resection of bladder tumor (TUR-BT). The sensitivity and specificity of the urine ThinPrep test was compared with that of conventional Cytospin cytology according to tumor number, size, pathological stage, grade, and recurrence. RESULTS: Of 3,085 subjects, bladder cancer was confirmed by TUR-BT in 379 subjects. The overall sensitivity of ThinPrep and Cytospin cytology was 60.9% and 59.9% in patients suspected of having bladder cancer, respectively. The overall specificity of ThinPrep and Cytospin cytology was 94.8% and 95.3% in patients suspected of having bladder cancer, respectively. The sensitivity of ThinPrep and Cytospin cytology was increased with increasing number, size, pathological stage, and grade, but there was no significant difference between the two tests. CONCLUSIONS: These results suggest that ThinPrep cytology has no advantage in the diagnosis of bladder cancer of a low grade or low stage.

A Comparative Study to Determine the Most Suitable Fixative for Immunocytochemical Staining of Cytospin Slides / 임상검사와정도관리

BACKGROUND: Fixation of cells is a critical procedure that can determine the success of immunocytochemical staining (ICC) of cytospin slides. In this study, we evaluated the efficacy of a number of fixatives to determine the ideal fixative for ICC of cytospin slides. METHODS: Sixteen patients with metastatic neoplasm in the body cavity were enrolled. Cytospin slides were prepared from each patient using 5 different fixatives (cold acetone, 95% ethanol, 1:1 methanol:ethanol, 3.7% formalin, and 3:1 methanol:acetone), and the suitability of each for use with Wright's stain was compared. For 4 of the samples, appropriate ICCs were performed using all 5 fixatives and the results were compared, while for 11 samples, only the first 3 fixatives were used for ICC. RESULTS: Using Wright stain, cold acetone, 95% ethanol, and 1:1 methanol:ethanol fixatives all showed similar efficacy when compared to the conventional methanol fixation method. However, the stain quality using 3.7% formalin or 3:1 methanol:acetone fixatives was poor due to deterioration of cell adhesion and distortion of cell morphology. Using ICC, cold acetone fixative showed stronger and more tumor-specific stainability than the 95% ethanol (decreased stainability in 6 stained slides, false positive staining of histiocytes/neutrophils in 4 stained slides, no staining of CD3 and terminal deoxynucleotidyl transferase [TdT]) and 1:1 methanol:ethanol fixatives (decreased stainability in 3 stained slides, false positive staining of histiocytes/neutrophils in 2 stained slides, no staining of CD3 and TdT). CONCLUSIONS: Cold acetone fixative was the most efficacious among the 5 fixatives tested in this study; therefore, it is the most appropriate fixative in the preparation of cytospin slides for ICC.

The Cytology for Leukemic Cells in Cerebrospinal Fluid; Comparison of Conventional Cytology with Liquid-Based Cytology

BACKGROUND: The cytological examination of cerebrospinal fluid (CSF) using conventional cytology with a cytocentrifuge (cytospin) is an important method for evaluating the involvement of leukemia in the CNS. Liquid-based cytology (LBC) is now a widely used cytological method not only for gynecological and non-gynecological specimens, but its application to CSF for the identification of leukemic cell has not yet been reported. In this study, we tried to compare conventional cytology with using a cytospin with LBC and Papanicolaou (Pap) staining. We also examined the modified LBC with Wright staining to assess whether this modified method can be useful for diagnosing Leukemia. METHODS: We studied 30 cases of CSF that were obtained from 16 patients, including 17 cases of acute myeloid leukemia, 12 cases of acute lymphoblastic leukemia and 1 case of diffuse large B cell lymphoma. We applied conventional cytology with a cytocentrifuge (cytospin), LBC with Pap staining and modified LBC with Wright staining. RESULTS: The morphological features of the LBC with Pap staining showed difficulty for interpretation when compared with conventional cytology with a cytospin, and mainly because of cellular shrinkage. The modified LBC with Wright staining showed good morphological features. CONCLUSIONS: We suggest that modified LBC with Wright staining may be useful for examining CSF.

Evaluation for Cytopreservability of Manual Liquid-Based Cytology Liqui-PREP(TM) and its Application to Cerebrospinal Fluid Cytology: Comparative Study with Cytospin / 대한세포병리학회지

Cerebrospinal fluid (CSF) cytology is an effective tool for evaluating diseases involving the central nervous system, but his technique is usually limited by its low cellularity and poor cellular preservation. Here we compared the manual liquid-base Liqui-PREPTM (LP) to the cytospin (CS) with using a mononuclear cell suspension and we applied both methods to the CSFs of pediatric leukemia patients. The cytopresevability, in terms of cell yield and cell size, and the clinical efficacy were evaluated. When 2000 and 4000 mononuclear cells were applied, LP was superior to CS for the cell yield, 16.8% vs 1.7% (P=0.001) and 26.2% vs 3.5% (P=0.002), respectively. The mean size of the smeared cells was 10.60 micrometer in the CS, 5.01 micrometer in the LP and 6.50 micrometer in the direct smear (DS), and the size ratio was 1.7 (CS to DS), 0.8(LP to DS) and 2.1 (CS to LP), respectively. As compared to the cells in the DS, the cells in the CS were significantly enlarged, but those in the LP were slightly shrunken. Upon application to 109 CSF samples, 4 were diagnosed as positive for leukemia (positive), 4 had atypical cells and 101 were negative by CS; 6 were positive, one had atypical cells and 102 were negative by LP. For six cases, in which 4 were positive for leukemia and 2 of 4 had atypical cells by CS, they were positive by LP and they were also confirmed as positive according to the follow-up study. Three cases diagnosed as atypical cells (two by CS and one by LP), were confirmed as negative. In conclusion, these results suggest that LP is superior to CS for the cytopresevability and for rendering a definite diagnosis of cerebrospinal fluid.

Comparision of Effectiveness between the ThinPrep(R) and the Cytospin Preparations of the Repeated Urine Cytology / 대한세포병리학회지

Once diagnosed as "cell paucity" or "atypia" by the cytospin (CS) preparation, this CS preparation does not secure a precise diagnosis by repeated testing alone. Although the ThinPrep(R) (TP) preparation is acknowledged to show increased cellularity, performing the screening tests for the cases that have enough cellularity, according to CS, raises issues for the cost-effectiveness. To obtain a more precise diagnosis through increasing the cellularity by performing TP, we selected the cases that were diagnosed as "cell paucity" or "atypia" by CS, but they required a more precise diagnosis, and the samples were processed via both CS and TP to compare the results. 11 patients diagnosed as "cell paucity" and 22 patients diagnosed as "atypia" by CS participated in this study. When the detection rate of atypical cells in both preparations with repeated urine cytology was compared, the overall detection rate of TP (16cases, 48.5%) was superior than that of CS (11cases, 33.3%), with statistical significance. The cellularity of both preparations was compared on repeated urine cytology; the general cellularity of TP (29cases, 87.9%) was higher than that of CS (20cases, 60.6%), but there was no statistical significance. Particularly, we repeated the TP for the 1 case that was diagnosed as "atypia" and we performed polyoma virus immunohistochemical staining, which confirmed polyoma virus. In conclusion, we can avoid obtaining negative diagnosis from cases with uncertain "atypia" or "cell paucity" by performing repeated TP testing.

The Diagnostic Utility of Mesothelial Markers in Distinguishing between Reactive Mesothelial Cell and Adenocarcinoma Cells in Serous Effusions with Cytospin Preparation / 대한세포병리학회지

Evaluation of serous effusions can include immunocytochemical stains that differentiate reactive mesothelial cell from adenocarcinoma cell. Among several positive mesothelial cell markers, we used desmin, CK5/6, WT1 and calretinin all known to have high sensitivity and specificity as selective mesothelial cell markers. We studied smears obtained with cytospin from 15 malignant and eight benign effusions. The mesothelial cells were positively stained by desmin, CK5/6, WT1 and calretinin in 60.9%, 29.1%, 26.7% and 56.5%, respectively among 8 benign and 15 malignant effusions; the adenocarcinoma cells were positively stained 6.7%, 13.3%, 1.0% and 0.0%, respectively among 15 malignant effusions. The percentage of positively stained mesothelial cells were somewhat lower for all antibodies compared to the results of previous studies. This was likely due to the differences in preparation methods and fixatives among studies. In conclusion, the use of desmin and calretinin were more valuable than CK5/6 and WT1 for distinguishing between reactive mesothelial cell and adenocarcinoma cells in serous effusion; however, choice of the proper preparation methods and fixatives are also important

Evaluation of the Usefulness of Immediate-Cytospin Wright-Stained Urine Cytology in the Screening and Monitoring of Bladder Cancer / 대한진단검사의학회지

BACKGROUND: The goal of this study is to assess the usefulness of immediate-cytospin Wright stain cytology with bladder tumor markers such as BTA and NMP22 in the screening and monitoring of bladder cancer. METHODS: We evaluated immediate-cytospin (cytospin 3, Shandon, UK) Wright stain and Papanicolau stain urinary cytology with the BTA stat (Bion Diagnostic Sciences, Inc., Redmond, WA, USA) and the NMP22 (Matritech, Newton, MA, USA) in 143 urine specimens for screening and monitoring of bladder cancer from September 2001 to May 2002. The cut-off decision level of NMP22 was determined by the receiver-operating characteristics (ROC) curve and those above the cut-off were classified as positive. The Wright and PAP stained urinary cytology results were reviewed by two experts. Negative and atypia cytology results were classified as negative, and suspicions of malignancy and malignancy results were classified as positive. RESULTS: The ROC curve analysis for NMP22 demonstrated optimal sensitivity and specificity (81.8% and 83.8%, respectively) with the cut-off decision level 11.0 IU/mL. Sensitivity, specificity, false positive rate, and false negative rate for the BTA stat were 62.5%, 92.9%, 47.4%, and 4.8%, respectively, and 81.8%, 83.8%, 55.0%, and 3.4%, respectively for the NMP22. Whereas, those values for immediate-cytospin Wright stain cytology were 90.9%, 98.4%, 16.7%, and 0.8%, respectively, showing a 100% concordance rate with Papanicolau stained cytology. The area under the ROC curve of the BTA stat, the NMP, and the immediate-cytospin Wright stain cytology was 0.78, 0.81, and 0.95, respectively. CONCLUSIONS: Immediate-cytospin Wright stain cytology was useful for the decrease in false positivity and false negativity for tumor markers such as the BTA stat and the NMP22 in screening and monitoring for bladder cancer.

Comparison of direct smear and cytospin of peritoneal fluid samplings in the diagnosis of gynecological malignancies.

While direct smear and cytospin of peritoneal fluid samplings have often been routinely prepared, their comparative usefulness and accuracy in the diagnosis of gynecological malignancies had not been evaluated in Siriraj Hospital Specimens obtained from 328 patients in whom clinical evidence raised the suspicion of gynecological malignancies were reviewed independently and without knowledge of the diagnosis. Direct smear gave better sensitivity and a lower false negative rate than cytospin (73.2% vs 69.6% and 26.8% vs 30.4%, respectively) but the difference was not statistically significant. Diagnostic accuracy can be improved by combining the two techniques. It can be concluded that direct smear is comparable to cytospin technique in diagnosing gynecological malignancies from peritoneal fluid samplings. Direct smear is the most useful method in places where cytospin is not available.