Large B-cell lymphomas: fine-needle aspiration plays an important role in initial diagnosis of cases which are falsely negative by flow cytometry

Large B-cell lymphomas (LBCLs) have significant false-negative results when immunophenotyped by flow cytometry (FC). To clarify the role fine-needle aspiration (FNA) in reducing this false-negative rate, 28 cases ultimately diagnosed as LBCL that had FNA as part of the workup and a negative FC were identified. We examined their clinical and cytologic features, in comparison with cases of LBCL with FNAs that were positive by FC. In 24/28 FC-negative cases (86%) a cytologic diagnosis of suspicious or positive for malignancy was rendered. We conclude that cytologic analysis is more sensitive than FC in the diagnosis of malignancy in FNA of LBCL, particularly in aspirates with low cellularity and/or low viability. Examination of cytospin preparations of the actual material analyzed by FC may provide an indication that an FC result is falsely negative. It is important to recognize the potential of false-negativity by FC of LBCLs when interpreting FNAs with features suggesting lymphoma.     

Comparison of three commonly used cytologic preparations in effusion immunocytochemistry.

Discrepant results in effusion immunocytochemistry are often the result of specimen processing. Smears, cytospins, cell blocks, and monolayer preparations have all been used in various published studies; thus, there is no consistency in the immunostaining process for cytology to compare with the surgical pathology "gold standard" results. We sought to evaluate optimal specimen preparation for the immunostaining of effusion samples. Fourteen reactive and 15 malignant effusion samples (various epithelial/mesothelial neoplasms) were each prepared in three forms: air-dried cytospins (postfixed in ethanol), formalin-fixed, paraffin-embedded cell blocks, and liquid-based thin-layer (ThinPrep, CYTYC, Boxborough, MA) processing. All slides were immunostained with antibodies commonly used in effusion cytology: HBME-1, calretinin, E-cadherin, BerEP4, B72.3, LeuM1, and CA19-9. Cytospin and ThinPrep samples performed in a similar manner: high background staining was encountered in 66% of cases, most evident in three-dimensional clusters of cells. In addition, membrane staining patterns were difficult to interpret. Cell blocks provided the best milieu for morphologic interpretation, with less background staining (only 17% of cases) and results that most closely approximated those reported in the surgical pathology literature. The cost per test for cell block immunocytochemistry was also the most economical for our laboratory.     

Use of ThinPrep monolayer technique and cytospin preparation in urine cytology: a comparative analysis

We compared the ThinPrep (TP) technique to the cytospin (CS) preparation in the cytological diagnosis of urine by processing 79 specimens by these two techniques. Ten cases were positive for malignancy (six high grade (HG)/carcinoma in situ; four low grade (LG) transitional cell carcinomas (TCC)). Forty-eight cases were within normal limits (59%) and 21 cases had atypical cytological features (19%). The TP technique was better in terms of a cleaner background with fewer obscuring inflammatory cells and blood and with a more even distribution of cells. In general, the cytomorphology was comparable in both techniques. However, in cases with malignancy, CS was relatively superior in the cytomorphologic details; in TP, the diagnostic cells were mostly dispersed as single cells with loss of architectural features and were difficult to find. Artifactual empty spaces and air-drying were more frequently present in TP. In cases contaminated with squamous cells, the urothelial cells were difficult to find in TP. Screening time was comparable for both techniques. In conclusion, to avoid false-negative diagnosis, CS would be complementary to the TP technique in malignant cases and, in particular, those with low cellularity.     

Fine-needle aspiration cytology of mammary fibroadenoma: a comparison of ThinPrep® and cytospin preparations

Mammary fibroadenoma (FA) is a lesion frequently sampled and diagnosed by fine‐needle aspiration (FNA). Accurate cytologic diagnosis of this common benign lesion is important as this can lead to non‐surgical, conservative management when breast imaging and clinical examination are concordant. In most instances, a confident diagnosis of FA is possible because of a characteristic cytologic appearance that includes hypercellularity, large epithelial cell groups, staghorn epithelial configurations, stromal fragments, and numerous background stripped nuclei. 1 , 2 Nevertheless, FAs can be diagnostically challenging because of shared cytomorphologic features with other benign lesions and low‐grade carcinoma. As such, FA is a well‐recognized source of false results on FNA cytology. 2 - 10 Furthermore, there are reports that newer thin layer cytopreparatory techniques, including the ThinPrep® (TP) system (Hologic Corp., Bedford, MA), alter the appearance of FA on FNA compared to conventional preparations and may compromise accurate cytologic diagnosis. 11 - 13 Diagn. Cytopathol. 2011;39:181–187. © 2010 Wiley‐Liss, Inc.     

Deoxyribonucleic acid content as an indicator of progression of squamous cell carcinogenesis in the esophagus: comparative analysis on imprint-cytospin and tissue section preparation

BACKGROUND: The purpose of this study was to explore the potential use of image analysis on tissue sections preparation as a predictive marker of early malignant changes during squamous cell (SC) carcinogenesis in the esophagus. Results of DNA ploidy quantification on formalin-fixed, paraffin-embedded tissue using two different techniques were compared: imprint-cytospin and 6 microm thick tissue sections preparation. METHODS: This retrospective study included 26 surgical specimens of squamous cell carcinoma (SCC) from patients who underwent surgery alone at the Department of Surgery in CHUV Hospital in Lausanne between January 1993 and December 2000. We analyzed 53 samples of healthy tissue, 43 tumors and 7 lymph node metastases. RESULTS: Diploid DNA histogram patterns were observed in all histologically healthy tissues, either distant or proximal to the lesion. Aneuploidy was observed in 34 (79%) of 43 carcinomas, namely 24 (75%) of 32 early squamous cell carcinomas and 10 (91%) of 11 advanced carcinomas. DNA content was similar in the different tumor stages, whether patients presented with single or multiple synchronous tumors. All lymph node metastases had similar DNA content as their primary tumor. CONCLUSIONS: Early malignant changes in the esophagus are associated with alteration in DNA content, and aneuploidy tends to correlate with progression of invasive SCC. A very good correlation between imprint-cytospin and tissue section analysis was observed. Although each method used here showed advantages and disadvantages; tissue sections preparation provided useful information on aberrant cell-cycle regulation and helped select the optimal treatment for the individual patient along with consideration of other clinical parameters.     

A comparison of the analysis of 3 types of body fluids using the XN-350 hematology analyzer versus light microscopy assessment

We evaluated the capacity of the XN-350 instrument to analyze 3 different types of body fluid samples under “body fluid mode.”The performance of XN-350 was evaluated in terms of precision, carryover, limit of blank, limit of detection, limit of quantification, and linearity. Cell enumeration and differential data produced by the XN-350 were compared to manual chamber counting results in 63 cerebrospinal fluid (CSF), 51 ascitic fluid, and 51 pleural fluid (PF) samples. Comparisons between XN-350 versus Cytospin data were also performed in PF samples.The precision, carry-over, limit of blank, and linearity of the XN-350 were acceptable. The limits of detection for white blood cells (WBCs) and red blood cells were 1.0/μL, and 1,000.0/μL, respectively; the corresponding limits of quantitation (LOQs) were 5.0/μL and 2,000.0/μL, respectively. The XN-350''s cell enumeration and differential counting correlated well with those of manual chamber counting for all 3 sample types (except for differential counting in CSF samples), particularly parameters involving monocytes (r = 0.33) and mononuclear cells (MO- body fluid [BF]; r = 0.26), as well as total cell (TC-BF) enumeration (r = 0.50) and WBC-BF (r = 0.50) in PF samples. The MO-BF in CSF samples differed significantly from manual chamber counting results, but neither TC-BF nor WBC-BF in PF samples did. The XN-350 also showed good correlations with Cytospin analyses for differential counting of neutrophils, lymphocytes, and monocytes in PF samples. The differential counting of eosinophils via the XN-350 and Cytospin were not significantly correlated, but the difference between them was not significant.The XN-350 is an acceptable alternative to manual fluid analysis. Samples with low cellularity around the LOQ should be checked manually. Moreover, manual differential counting should be performed on CSF samples, particularity those with low cell numbers.     

Utility of immunocytochemistry in diagnosing leptomeningeal metastases from an intrahepatic cholangiocarcinoma

Isolated spinal leptomeningeal metastases (LMM) without brain metastases are infrequent, accounting for about 1% of all solid tumors. In LMM, cerebrospinal fluid (CSF) analyses are mostly abnormal. Demonstrations of intrathecal tumor markers are highly suggestive, but only a positive cytology is diagnostic. The initial CSF cytology can give a false negative result in up to 40–50% of patients with pathologically proven LMM on autopsy. We report a case of intrahepatic cholangiocarcinoma with spinal LMM confirmed using cytokeratin7 and pancytokeratin (AE1/AE3) immunocytochemical studies on paucicellular cerebrospinal fluid cytospin preparation. Given the paucicellularity of the smears and difficult morphologic categorization, immunocytochemistry is vital for confirmatory diagnosis and can help reduce false negative results. To the best of our knowledgethis is the first case report of cytologically confirmed LMM from an intrahepatic cholangiocarcinoma while the patient was undergoing treatment. Diagn. Cytopathol. 2014;42:54–57. © 2013 Wiley Periodicals, Inc.