Characterizing the Effect of Processing Technique and Solution Type on Cytomorphology Using Liquid-Based Cytology

<b><i>Introduction:</i></b> Liquid-based cytology (LBC) is increasingly used for nongynecologic applications. However, the cytological preparation of LBC specimens is influenced by the processing technique and the preservative used. In this study, the influence of the processing techniques and preservatives on cell morphology was examined mathematically and statistically. <b><i>Methods:</i></b> Cytological specimens were prepared using the ThinPrep (TP), SurePath (SP), and AutoSmear methods, with 5 different preservative solutions. The cytoplasmic and nuclear areas of Papanicolaou-stained specimens were measured for all samples. <b><i>Results:</i></b> The cytoplasmic and nuclear areas were smaller in cells prepared using the 2 LBC methods, compared to that prepared using the AutoSmear method, irrespective of the preservative used. The cytoplasmic and nuclear areas of cells prepared using the SP method were smaller than those of cells prepared using the TP method, irrespective of the preservative used. There were fewer differences among the cytoplasmic areas of cells prepared with different preservative solutions using the TP method; however, the cytoplasmic areas of cells prepared using the SP method changed with the preservative solution used. <b><i>Conclusions:</i></b> The most significant difference affecting the cytoplasmic and nuclear areas was the processing technique. The TP method increased the cytoplasmic and nuclear areas, while the methanol-based PreservCyt solution enabled the highest enlargement of the cell. LBC is a superior preparation technique for standardization of the specimens. Our results offer a better understanding of methods suitable for specimen preparation for developing precision AI-based diagnosis in cytology.

EVALUATION OF CELL BLOCK TECHNIQUE VERSUS CONVENTIONAL CYTOLOGY ON BRONCHOSCOPY GUIDED NEEDLE ASPIRATION/ BRUSH CYTOLOGY FOR DIAGNOSIS OF LUNG CANCER

INTRODUCTION: Bronchoscopy is a safe and effective method for diagnosing lung carcinomas with a variation in the diagnostic yield with different bronchoscopy guided procedures. Cell block technique has shown an addition cases positivity in diagnosing carcinomas as compared to the conventional method. AIM: The present study was aimed to evaluate the diagnostic utility of cell block technique on Bronchoscopy guided needle aspiration/ Brush and also to compare cytological preparation with cell block. MATERIAL AND METHODS: A total of 50 cases were included in the study that was suspected to be having lung carcinoma. These patients went under bronchoscope guided aspirations (TBNA, EBNA, and Brush). Smears were immediately made for conventional cytology study and well as in another aliquot samples were collected to prepare cell blocks following which H&E staining was done. RESULTS: Out of 50 cases, 8 cases came out to be negative on conventional smears and when compared with cell block technique 4 additional cases came out to be positive who were negative on conventional smears. The diagnosis were compared with histopathology biopsies keeping it as a gold standard and results on cell block techniques were conrmed to be true. CONCLUSION: Out of 50 cases, an additional 4 more cases were diagnosed malignant by using the cell blocks technique but there were few drawbacks with cell block technique. In few of the cases on cell block, cellularity was very less, cells morphology was also not very clear and some showed cells entrapped in a clusters. The conclusion made out of this study is that cell block technique is more accurate than the cytological smears and when used in combination diagnostic efcacy will be improved.

APPLICATION OF THE CYTOLOGICAL METHOD OF ASSESSMENT OF THE WOUND PROCESS IN GUNSHOT INJURIES OF THE LIMB

The article presents the results of studies devoted to the substantiation of the use of the cytological express method for assessing the wound process in patients with gunshot injuries of the extremities. In the course of the study, wound exudate was taken from the area of the most altered tissues in the projection of the wound channel, followed by preparation of preparations. The first and second group of preparations were stained according to Romanovsky-Giemsa and Alekseev's method in the cytological laboratory. The preparations of the third group were stained with the May-Grunwald dye in the preoperative room. Comparative assessment was carried out using the developed color quality index of the cytological preparation, based on a point scale. It was found that the quality of preparations stained with the May-Grunwald dye was lower according to a number of criteria (completeness of staining of cells and cell structures, the presence of artifacts, focusing resolution) in comparison with preparations stained by the laboratory method. Despite this fact, staining with eosin methylene blue according to May-Grunwald made it possible to establish the type of cytogram and assess the dynamics of the wound process within a short time (no more than 7-8 minutes) directly in the preoperative room. The advantages of this method are: the ability to get a conclusion in the shortest possible time, learnability, repeatability, the absence of the need to involve a specialist in laboratory diagnostics. This technique is a surgeon's working tool for practical targeting of the wound, taking into account an objective assessment of the wound process.

Placental Site Plaque of the Uterine Cervix Misinterpreted as Low-Grade Squamous Intraepithelial Lesion in Liquid-Based Cervicovaginal Cytology: Usefulness of Inhibin-α Immunocytochemistry

Cytological features of placental site plaques in liquid-based cervicovaginal preparations have been seldom documented in the literature. We present a rare case of endocervical placental site plaque misinterpreted as a low-grade squamous intraepithelial lesion in a liquid-based cytological preparation. A 32-year-old woman with polycystic ovarian syndrome gave birth 7 months previously. After delivery, she was diagnosed with cervical low-grade squamous intraepithelial lesion during routine cytological examination. Cytologically, many atypical cells showed large hyperchromatic nuclei with irregular membranes. The perinuclear cytoplasmic clearing closely resembled koilocytosis. Histologically, the endocervix showed typical histological features of a placental site plaque. Immunohistochemically, the trophoblasts were positive for p63, CD10, and inhibin-α but negative for p16. Based on genotyping, both the cytological and biopsied specimens tested negative for human papillomavirus. We re-examined the liquid-based preparation cytology slides thoroughly and concluded that the atypical cells initially misinterpreted as low-grade squamous intraepithelial lesion were actually trophoblasts. Immunocytochemical staining revealed uniform cytoplasmic inhibin-α expression in the trophoblasts. In summary, we demonstrated that endocervical placental site plaques can mimic low-grade squamous intraepithelial lesions in liquid-based cytological preparations. Immunocytochemical staining results and negative results on human papillomavirus genotyping further support that atypical cells resembling koilocytes are trophoblasts obtained from the placental site plaque.

Comparative Analysis of Rhino-Cytological Specimens with Image Analysis and Deep Learning Techniques

Cytological study of the nasal mucosa (also known as rhino-cytology) represents an important diagnostic aid that allows highlighting of the presence of some types of rhinitis through the analysis of cellular features visible under a microscope. Nowadays, the automated detection and classification of cells benefit from the capacity of deep learning techniques in processing digital images of the cytological preparation. Even though the results of such automatic systems need to be validated by a specialized rhino-cytologist, this technology represents a valid support that aims at increasing the accuracy of the analysis while reducing the required time and effort. The quality of the rhino-cytological preparation, which is clearly important for the microscope observation phase, is also fundamental for the automatic classification process. In fact, the slide-preparing technique turns out to be a crucial factor among the multiple ones that may modify the morphological and chromatic characteristics of the cells. This paper aims to investigate the possible differences between direct smear (SM) and cytological centrifugation (CYT) slide-preparation techniques, in order to preserve image quality during the observation and cell classification phases in rhino-cytology. Firstly, a comparative study based on image analysis techniques has been put forward. The extraction of densitometric and morphometric features has made it possible to quantify and describe the spatial distribution of the cells in the field images observed under the microscope. Statistical analysis of the distribution of these features has been used to evaluate the degree of similarity between images acquired from SM and CYT slides. The results prove an important difference in the observation process of the cells prepared with the above-mentioned techniques, with reference to cell density and spatial distribution: the analysis of CYT slides has been more difficult than of the SM ones due to the spatial distribution of the cells, which results in a lower cell density than the SM slides. As a marginal part of this study, a performance assessment of the computer-aided diagnosis (CAD) system called Rhino-cyt has also been carried out on both groups of image slide types.

Preservation of cellular nano-architecture by the process of chemical fixation for nanopathology

Transformation in chromatin organization is one of the most universal markers of carcinogenesis. Microscale chromatin alterations have been a staple of histopathological diagnosis of neoplasia, and nanoscale alterations have emerged as a promising marker for cancer prognostication and the detection of predysplastic changes. While numerous methods have been developed to detect these alterations, most methods for sample preparation remain largely validated via conventional microscopy, and have not been examined with nanoscale sensitive imaging techniques. For these nanoscale sensitive techniques to become standard of care screening tools, new histological protocols must be developed that preserve nanoscale information. Partial Wave Spectroscopic (PWS) microscopy has recently emerged as a novel imaging technique sensitive to length scales ranging between 20 and 200 nanometers. As a label-free, high-throughput, and non-invasive imaging technique, PWS microscopy offers many advantages for risk stratification of early cancer, and is an ideal tool to quantify structural information during sample preparation. Therefore, in this work we applied PWS microscopy to systematically evaluate the effects of cytological preparation on the nanoscales changes of chromatin using two cell line models: Hela cells differentially treated with daunorubicin and TP53 differentially mutated ovarian carcinoma cells. Notably, we show that existing cytological preparation can be modified in order to maintain clinically relevant nanoscopic differences, paving the way for the emerging field of nanopathology.

A Prospective Study of Soft Tissue Tumors Histocytopathology Correlation

Background. Soft tissue tumors are defined as nonepithelial extraskeletal tissue of the body exclusive of the reticuloendothelial system, glia, and supporting tissue of various parenchymal organs. The absence of recognizable tissue architectural patterns in cytological preparation makes diagnosis by FNAC more difficult. Aims. To assess the utility of FNAC in diagnosing soft tissue tumors and to determine their patterns compared with with the respective histopathology results. Materials and Methods. 150 cases of soft tissue tumors were included in this study for cytologic and histologic correlation. FNAC air dried smears were stained with Giemsa stain and 95% ethanol fixed smears were stained with Papanicolaou stain. The smears were studied for cytological diagnosis and were categorized as benign, suspicious of malignancy, and malignant along with specific subtyping of the lesion. All diagnostic FNAC results were compared for diagnostic concordance using histology results as the “gold standard.” Results. The sensitivity, specificity, positive predictive value, negative predictive value, and efficiency were 70%, 100%, 97.90%, 100%, and 98%, respectively. P value was <0.0001 which shows statistically extreme significant correlation. Conclusion. FNAC is a very important preliminary diagnostic tool in palpable soft tissue lumps with high degree of correlation with the final histopathology report.