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Liquid-based preparation (LBP) cytology is commonly used in most laboratories these days due to its convenience and reliable results for the cervical cancer screening program. The PathTezt™ Liquid-based Pap smear is a second-generation LBP, which uses a filter-based concentration technique in processing the sample., Objective: This study was done to evaluate the cellular fixation, morphology, quality of smear in gynae cytology, and diagnostic interpretation of cervical cytological smears produced by the PathTezt liquid-based processor., Materials and Methods: A total of 400 pap smear samples were taken and processed using the PathTezt 2000 processor. The slides were evaluated in terms of sample adequacy, percentage of the circle covered by epithelial cells, cellular distribution, obscuring factors, and cell fixation., Results: About 95.25% (381) of the samples were satisfactory for the evaluation. In 19 (4.75%) of the samples, epithelial cells covered less than 50% of the circle. A sample with good cellular distribution was seen in 92% of the cases, while 354 (88.5%) samples showed minimal inflammatory background. Almost all the smears (95.75%) had no erythrocytes in the background. All smears showed good quality fixation features toward nuclear, cytoplasm, and microorganisms. The total performance rate was 99%., Conclusion: Although the PathTezt liquid-based processor is still new compared to other first-generation LBP, the smears produced by this method were of high quality and it was cost-effective.

Objective: The aim of the study was to propose a method using saline lubrication and two glass slides to reduce the proportion of inadequate Pap smears., Materials and Methods: This was a retrospective study of patients at a medical center in eastern Taiwan that performs 5000-6000 Pap smears annually. The extracted data only detailed the number and percentage of inadequate Pap smears. We applied two modifications to the conventional Pap smear technique. The first modification was lubricating the speculum with normal saline instead of jelly. The second modification was performing the smear on two glass slides instead of just one. We used the modified technique beginning in January 2017. Therefore, we collected data from 2016 (before the modified technique was employed) and 2018 (after the modified technique was employed). The categorical data are presented as numbers (percentages). The differences in the number and percentage of inadequate smears resulting from both techniques were assessed using the Chi-square test., Results: During 2016 and 2018, 28 and 2 women received inadequate Pap smears among the total of 594 and 613 women who received Pap smears, respectively. The proportion of inadequate Pap smears was 4.71% and 0.33% in 2016 and 2018, respectively (P < 0.001)., Conclusions: The use of this modified technique effectively reduced the percentage of inadequate Pap smears., Competing Interests: Declaration of competing interest None to declare., (Copyright © 2020. Published by Elsevier B.V.)

Background: The primary high-risk human papillomavirus (hrHPV) assays in the United States are the cobas (Roche) and the Aptima (Hologic). The cobas assay detects hrHPV by DNA analysis while the Aptima detects messenger RNA (mRNA) oncogenic transcripts. As the Aptima assay identifies oncogenic expression, it should have a lower rate of hrHPV and genotype detection., Methods: The Kaiser Permanente Regional Reference Laboratory in Denver, Colorado changed its hrHPV assay from the cobas to the Aptima assay. The rates of hrHPV detection and genotyping were compared over successive six-month periods., Results: The overall hrHPV detection rates by the two platforms were similar (9.5% versus 9.1%) and not statistically different. For genotyping, the HPV 16 rate by the cobas was 1.6% and by the Aptima it was 1.1%. These differences were statistically different with the Aptima detecting nearly one-third less HPV 16 infections. With the HPV 18 and HPV 18/45, there was a slightly higher detection rate of HPV 18/45 by the Aptima platform (0.5% versus 0.9%) and this was statistically significant., Conclusion: While HPV 16 represents a low percentage of hrHPV infections, it was detected significantly less by the Aptima assay compared to the cobas assay. This has been previously reported, although not highlighted. Given the test methodologies, one would expect the Aptima to detect less HPV 16. This difference appears to be mainly due to a significantly increased number of non-oncogenic HPV 16 infections detected by the cobas test as there were no differences in HPV 16 detection rates in the high-grade squamous intraepithelial lesions indicating that the two tests have similar sensitivities for oncogenic HPV 16., (© 2018 Wiley Periodicals, Inc.)

Background: Whole-slide imaging in cytology is limited when glass slides are digitized without z-stacks for focusing. Different vendors have started to provide z-stacking solutions to overcome this limitation. The Panoptiq imaging system allows users to create digital files combining low-magnification panoramic images with regions of interest (ROIs) that are imaged with high-magnification z-stacks. The aim of this study was to compare such panoramic images with conventional whole-slide images and glass slides for the tasks of screening and interpretation in cytopathology., Methods: Thirty glass slides, including 10 ThinPrep Papanicolaou tests and 20 nongynecologic cytology cases, were digitized with an Olympus BX45 integrated microscope with an attached Prosilica GT camera. ViewsIQ software was used for image acquisition and viewing. These glass slides were also scanned on an Aperio ScanScope XT at ×40 (0.25 μm/pixel) with 1 z-plane and were viewed with ImageScope software. Digital and glass sides were screened and dotted/annotated by a cytotechnologist and were subsequently reviewed by 3 cytopathologists. For panoramic images, the cytotechnologist manually created digital maps and selected representative ROIs to generate z-stacks at a higher magnification. After 3-week washout periods, panoramic images were compared with Aperio digital slides and glass slides., Results: The Panoptiq system permitted fine focusing of thick smears and cell clusters. In comparison with glass slides, the average screening times were 5.5 and 1.8 times longer with Panoptiq and Aperio images, respectively, but this improved with user experience. There was no statistical difference in diagnostic concordance between all 3 modalities. Users' diagnostic confidence was also similar for all modalities., Conclusions: The Aperio whole-slide scanner with 1 z-plane scanning and the Panoptiq imaging system with z-stacking are both suitable for cytopathology screening and interpretation. However, ROI z-stacks do offer a superior mechanism for overcoming focusing problems commonly encountered with digital cytology slides. Unlike whole-slide imaging, the acquisition of representative z-stack images with the Panoptiq system requires a trained cytologist to create digital files. Cancer Cytopathol 2017;125:701-9. © 2017 American Cancer Society., (© 2017 American Cancer Society.)

Cervical cancer screening based on the Papanicolaou (Pap) test is a widely applied but not always efficient practice for detecting Human Papillomavirus (HPV) mediated lesions, partially due to a non-systematic and inadequate screening process. Our aim was to introduce an inexpensive easy-to-use direct screening platform for improved detection of abnormal cells indicative of underlying cervical neoplasia as well as persisting HPV infection. By employing a novel, efficient technique of laser-based micromachining, we achieved the fabrication of spatial grids on commercially available coverslips allowing visual segmentation of the slide for efficient screening. Abnormal and formerly diagnosed as negative for intraepithelial lesion or malignancy (NILM) Pap test slides (n = 200) were analyzed by conventional and grid-based screening. Grid-based microscopy led to a more reliable diagnosis compared to the conventional by identifying an increased number of abnormal cells (P = 0.001). It decreased borderline ASCUS, AGC diagnosis, increasing LSIL, HSIL and in situ AdenoCa detection rates closely related with biopsy (P = 0.015; kappa = 0.978). Concerning the set of NILM diagnoses in rapid re-screening, the method upgraded six cases (n = 6) to LSIL (P = 0.001). The proposed technical solution offers a calibration and orientation visual aid during the on-site screening process providing significant advantages compared to expensive digital imaging techniques., (© 2016 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.)

Background: In the fine needle aspiration cytology (FNAC) for tumors of the breast, evaluation is frequently difficult because of the thick-layered cell clusters and blood inclusion. Such problems may be resolved by the returned cell block method, but its use has not spread worldwide. Here, we examined the application of the returned cell block method to cases involving difficulty in the evaluation of FNAC to diagnose tumors of the breast., Methods: In Juntendo University Nerima Hospital, there were 22 cases which were difficult to diagnose by Papanicolaou stain only, and they underwent additional examination using the returned cell block method (cell block from a Papanicolaou staining smear on a glass slide). The usefulness of the returned cell block method in these cases was examined., Results: Among the 22 cases, a correct diagnosis was facilitated in 20 cases using the returned cell block method. In 16 of the 20 cases, the difficulty in FNAC was because of thick-layered cell clusters (12 cases) and blood inclusion (four cases). Among the 12 cases with difficulty because of the thick-layered cell clusters, 10 cases (83%) comprised intraductal papilloma (six cases) and intraductal papillary carcinoma (four cases). Papilloma and papillary carcinoma were correctly diagnosed by the addition of histological images and immunostaining of myoepithelial cells using the returned cell block method., Conclusion: The application of the returned cell block method is useful for precise evaluation of the cytological diagnosis of tumors of the breast, especially papillary lesions. Diagn. Cytopathol. 2016;44:505-511. © 2016 The Authors Diagnostic Cytopathology Published by Wiley Periodicals, Inc., (© 2016 The Authors Diagnostic Cytopathology Published by Wiley Periodicals, Inc.)

Background: Although glacial acetic acid (GAA) treatment of bloody cervical samples has reduced the rate of unsatisfactory Pap Tests, recent studies suggest that it may negatively impact high-risk (hr)-HPV test results. The objectives of this study were to compare the levels of genomic DNA between GAA treated and nontreated ThinPrep(®) samples using the Hologic(®) Cervista(®) HPV-HR assay and to compare the adequacy of the ThinPrep(®) Pap Test between aliquoted and nonaliquoted samples., Methods: Prior to GAA treatment, 2.5 ml of the cervical sample was prealiquoted from 102 bloody ThinPrep(®) vials. Both GAA treated and nontreated samples were analyzed for hr-HPV using the Cervista(®) HPV HR assay. The levels of genomic DNA were measured and compared between these samples. In addition, ThinPrep(®) Pap Test adequacy rates were calculated and compared on aliquoted and nonaliquoted cervical samples., Results: Of the 102 cervical samples, 95 (93%) nontreated aliquots contained satisfactory levels of genomic DNA as compared to 36 (35%) GAA-treated samples (p < 0.00001). Ninety-nine (97%) aliquoted cervical samples were satisfactory for cytologic evaluation as compared to 1,326 (96%) GAA treated samples from 2013 (nonaliquoted); not statistically significant (p = 0.7968)., Conclusion: The levels of genomic DNA were significantly decreased in GAA treated vs non-treated TP samples. Aliquoting from the TP sample prior to treatment with GAA enables accurate measurement of DNA without affecting the adequacy of the TP cytology slide., (© 2016 Wiley Periodicals, Inc.)

Background: From 2010 to 2014, approximately 2 million Pap smears from HIV-infected women were submitted to the South African National Health Laboratory Services (NHLS) through the national cervical cancer screening programme. The objective of this analysis was to determine whether using the plastic Cervex brush ("broom") would be a cost-effective approach to improve cytology specimen quality as compared to the wooden spatula used currently., Methods: A decision analysis model was built using the expected adequacy rates for samples collected with the spatula (<$0.02) and broom ($0.23) and the probability of detecting cervical dysplasia. NHLS data was used for testing volumes and rates of HIV-positivity, suitability of specimens, and presence of endocervical cells. Expected positivity of Pap smears in HIV-infected women (73 %), odds ratios of the effectiveness of the broom (OR: 1.57), and improved sensitivity when endocervical cells present (OR: 1.89) are from literature. NHLS costs were used for the collection devices and conventional cytology ($4.89). Cost of clinic visit is from WHO CHOICE ($8.36)., Results: In 2010, 80 % of specimens submitted to NHLS were adequate for evaluation; in 2014, only 54 % met the same criteria. For HIV-infected women, according to the guidelines model, using the wooden spatula costs $6.25 million per year, $16.79 per woman tested. Under intended practice, for each additional HSIL case detected among HIV-infected women, the South African cervical cancer screening programme could save $13.64 (95 % CI: $13.52 to $13.76) by using the broom as its standard of care collection device through increased collection of endocervical cells and consequent reduction in repeat Pap smears., Conclusion: Under a wide range of parameters tested using a simulation model, the more expensive plastic broom could save the South African cervical cancer screening programme money and increase detection of high-grade cervical dysplasia in HIV-infected women compared to the current wooden spatula.

Proflavine hemisulfate, an acridine-derived fluorescent dye, can be used as a rapid stain for cytologic examination of biological specimens. Proflavine fluorescently stains cell nuclei and cytoplasmic structures, owing to its small amphipathic structure and ability to intercalate DNA. In this manuscript, we demonstrated the use of proflavine as a rapid cytologic dye on a number of specimens, including normal exfoliated oral squamous cells, cultured human oral squamous carcinoma cells, and leukocytes derived from whole blood specimens using a custom-built, portable, LED-illuminated fluorescence microscope. No incubation time was needed after suspending cells in 0.01% (w/v) proflavine diluted in saline. Images of proflavine stained oral cells had clearly visible nuclei as well as granular cytoplasm, while stained leukocytes exhibited bright nuclei, and highlighted the multilobar nature of nuclei in neutrophils. We also demonstrated the utility of quantitative analysis of digital images of proflavine stained cells, which can be used to detect significant morphological differences between different cell types. Proflavine stained oral cells have well-defined nuclei and cell membranes which allowed for quantitative analysis of nuclear to cytoplasmic ratios, as well as image texture analysis to extract quantitative image features.

As digital imaging is becoming a fundamental part of medical and biomedical research, the demand for computer-based evaluation using advanced image analysis is becoming an integral part of many research projects. A common problem when developing new image analysis algorithms is the need of large datasets with ground truth on which the algorithms can be tested and optimized. Generating such datasets is often tedious and introduces subjectivity and interindividual and intraindividual variations. An alternative to manually created ground-truth data is to generate synthetic images where the ground truth is known. The challenge then is to make the images sufficiently similar to the real ones to be useful in algorithm development. One of the first and most widely studied medical image analysis tasks is to automate screening for cervical cancer through Pap-smear analysis. As part of an effort to develop a new generation cervical cancer screening system, we have developed a framework for the creation of realistic synthetic bright-field microscopy images that can be used for algorithm development and benchmarking. The resulting framework has been assessed through a visual evaluation by experts with extensive experience of Pap-smear images. The results show that images produced using our described methods are realistic enough to be mistaken for real microscopy images. The developed simulation framework is very flexible and can be modified to mimic many other types of bright-field microscopy images., (© 2015 The Authors. Published by Wiley Periodicals, Inc. on behalf of ISAC.)

Background: Evaluate the performance of the Focalpoint system in identifying and classifying cervical cytology alterations from samples collected from patients treated with Radiotherapy (RT)., Methods: The reproducibility of manual and automated screening by cytotechnologists using the BD Focalpoint GS Imaging System was examined. Samples were collected from May 2010 to August 2011., Results: A total of 378 treated with RT and 8,967 cytology samples from patients without previous RT, were evaluated. The kappa values for cytological diagnoses read manually and automated in cases without previous RT were as follows: < ASC-H vs. ≥ ASC-H = 0.71; < LSIL vs. ≥ LSIL = 0.66; and < HSIL vs. ≥ HSIL = 0.67. The kappa for cytological diagnoses in post-RT women have showed: < ASC-H vs. ≥ ASC-H = 0.71; < LSIL vs. ≥ LSIL = 0.65; < HSIL vs. ≥ HSIL = 0.57., Conclusions: There was no significant difference among the kappa values we found. Post-RT cytology showed small diagnostic agreement between manual and automated examination., Virtual Slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000&#95;2014&#95;231.

Previous prospective studies of automated assisted gynecologic screening devices have used a panel of experts for truth determination. We sought to determine the value of this practice. The relative sensitivity of the devices compared with manual screening was calculated using an expert panel for truth determination and compared using likelihood ratios to the relative sensitivity assuming all abnormal cases were truly abnormal. These results show that expert panel review has no significant effect on relative sensitivity at the threshold of ASCUS+ but may have an effect at HSIL+. Trials without expert consensus review may be compared to those with expert consensus review at the threshold of ASCUS+ but may not be reliable at the threshold of HSIL+ without additional confirmatory data., (Copyright © 2013 Wiley Periodicals, Inc.)

Objective: It was the aim of this study to assess the utility of the manual liquid-based cytology (LBC) product VitroPrep™ Cytology Processing Kit (ChemQ Bioscience LLC, Research Triangle Park, N.C., USA)., Study Design: This is a descriptive pilot study. Women underwent cervical sampling processed by the ThinPrep™ automated LBC system followed by cervical sampling for the VitroPrep manual system. The following criteria were assessed on a scale of 1-5 (1 = unsatisfactory, 2 = borderline, 3 = acceptable, 4 = good, 5 = excellent): monolayer cell adhesion, overall cellularity, background clarity, preservation of cellular morphology, red cell lysis, and elimination of mucus/debris. Cytological diagnosis was compared to results from ThinPrep samples. In addition, VitroPrep samples were taken prior to conization procedures and compared to pathology results. Descriptive statistics were performed., Results: Forty-two of 47 women who underwent dual cytologic sampling had satisfactory samples. All scores were 3-5, with >90% graded 4-5. The VitroPrep diagnosis correlated with the ThinPrep diagnosis in 90% (38/42) of cases. All specimens obtained from 15 women prior to conization were satisfactory and correlated abnormal cytologic findings with cervical intraepithelial neoplasia 1-3 pathology., Conclusions: The VitroPrep Cytology Processing Kit was able to provide adequate specimens for evaluation and diagnosis. This low-cost processing kit may provide a useful alternative in settings where automated LBC systems may not be feasible., (© 2014 S. Karger AG, Basel.)

Objective: A double-blind study was conducted to compare the performance of the new BestPrep® (CellSolutions) liquid-based thin-layer Papanicolaou (Pap) test with ThinPrep® (Hologic)., Study Design: Samples from the study patients (n = 105) were collected twice in the same encounter with the ThinPrep sample always taken first and the BestPrep sample collected second. Slides were prepared according to both manufacturers' protocols and evaluated using manual microscopic review and the BestCyte® cell sorter imaging system (CellSolutions). Diagnostic truth for each case was determined by independent manual review of both slides by multiple pathologists and histology when available. The presence of atypical squamous cells of undetermined significance was the threshold for positive for sensitivity and specificity calculations., Results: BestPrep and ThinPrep, by manual review, had sensitivities for high-grade squamous intraepithelial lesion (HSIL) cases of 100 and 95.6%, respectively. Using the BestCyte cell sorter, both had 100% sensitivity. For the same HSIL cases, the digene HC2 high-risk human papillomavirus DNA test had sensitivities of 100% (BestPrep) and 95.6% (ThinPrep). Specificities were 71.4% (BestPrep) and 54.8% (ThinPrep)., Conclusions: BestPrep was equivalent to ThinPrep for manual review even though BestPrep was always the second sample collected. The BestCyte cell sorter provides a practical alternative to manual review for both BestPrep and ThinPrep slides., (© 2014 S. Karger AG, Basel.)