Set Of Smears Research Articles

CONVENTIONAL CYTOLOGY VERSUS LIQUID BASED CYTOLOGY FOR EVALUATION OF CERVICAL SMEARS – A STUDY OF 300 CASES

BACKGROUND: Over the years, different studies conducted have established the advantages of liquid based cytology(LBC) over conventional preparations(CP) in cervical smears screening. However, the high cost of machinery required for the preparations still limits its usage in low-resource countries. This study aims to evaluate the performance of a low cost LBC method, EziPREP™ technique against conventional preparations for cervical screening. MATERIALS & METHOD: This was a cross sectional study comprising of 300 consequtive cervical samples over a period of 2 year. CP smears were prepared as per the standard technique using ayre’s spatula while for LBC sample was obtained on endocervical brush followed by detaching the head of brush into the fixative vials of LBC. The LBC samples were processed as per the manufacturer's protocol. Both CP and LBC smears were stained using standard Papanicolaou stain & H & E stain protocol. Slides were analysed by two pathologists independently. RESULTS: The unsatisfactory rate for CP was 5%, main cause being obscuring haemorrhage and inflammation, whereas on LBC samples it was 1.4%, owing to inadequate cellular material. Uniform distribution was more in LBC smears, with minimal overlapping. The morphological details were comparable in both sets of smears. The pathogenic organisms, like Gardenella(31%) & Candida(20%) were detected in more CP smears, Trichomonas was seen equally in both smears(3%). There was a 98% concordance in cytologic diagnosis between CP and LBC smears. Screening time was notably reduced in LBC smears (3±1minutes vs 5±1 minutes for CP) CONCLUSION: This low-cost LBC method holds promise in providing monolayered cervical smears with a clear background. It might carve the way for inclusion of liquid based cytology methods for cervical cancer screening on a larger scale in the low resourse countries.

Study the estrogen (ER) and progesterone (PR) receptor in case of primary and metastatic breast cancer

Background: Worldwide, breast cancer is the most common invasive cancer in women.It comprises 22.9% of invasive cancers in women and 16% of all female cancers. It accounts for 15% of cancer deaths worldwide.Objective: to study the expression of ER/PR on breast cancer by immunocytochemistry on fine needle aspiration smears. Material and Methods: A prospective study was carried out on 120 cases of breast lump, referred from various clinical Departments to cytopathology lab of a tertiary care center in North India, during September 2018 to March 2020. Two sets of Fine Needle Aspirate smears were obtained from breast lump and palpable lymph node, if present. Both sets of smear were air dried and fixed in propyl alcohol for 1-2 hours.First set of smear was used for cytology, in which cytology was positive, the second set of smear were packed in aluminum foil and preserved at -20 degree Celsius. Slides were then stained for ER and PR receptor. Later on when patient undergone surgery findings were correlated with histopathology. Results : The most common symptom was breast lump (87.5%) with tumour size of 2.0-5.0 cm. ER were positive in 65.0% cases, PR were positive in 57.5% and ER+/PR+ in 55.0% of cases.

Comparative cytopathologic evaluation using acridine orange with Papanicolaou stain in psychoactive substances abusers with potentially malignant and malignant disorders

Introduction:Oral cancer is a major health problem and its early detection is advantageous for therapeutic purposes. According to available evidence, the risks of oral malignancies increase with the usage of tobacco and other psychoactive substances (PSs). The present study showed expression pattern of nuclear and cytoplasmic changes from normal individuals without habit to oral potentially malignant disorders (OPMD) and oral squamous cell carcinoma (OSCC) in PS abusers with the help of fluorescence acridine orange (AO) stain and Papanicolaou (PAP) stain.Aim and Objectives:This study aimed to investigate and compare diagnostic efficacy of fluorescence microscopic evaluation of AO stain in cytological smears with PAP staining under light microscopy in PS abusers having oral potentially malignant and malignant lesions.Materials and Methods:Oral smears from 120 individuals among which 40 from potentially malignant disorders, 40 from oral malignancy and 40 normal buccal mucosa smears were prepared. One set of smears was stained by AO staining and the other by PAP staining and examined under fluorescence and light microscope, respectively. The results of both the stainings were evaluated by grading cytology smears in class-I to class-V cytology.Results:The AO fluorescence stain reliably demonstrated malignant cells based on the differential fluorescence. The efficacy of AO fluorescence stain was higher than PAP stain in screening of oral lesions suspicious of malignancy. The sensitivity of PAP staining and AO staining is 57.50% and 61.25%, respectively.Conclusion:As compared to PAP staining method, fluorescent AO method is more effective in screening of OPMD and OSCC in PS abusers.

Analysis of Papanicolaou stain on peripheral smear compared to Leishman's stain: A prospective study

Aim: The aim of the study is to know the exact effect of Papanicolaou (Pap) staining on the peripheral smear while compared to Leishman's stain on the peripheral smear. Background: Pap staining is discovered and usually used for wet-fixed cytological smears. As a differential stain and using hematoxylin and eosin as a part of the staining solutions, Pap stains also have a principle of acidic and basic pH and contents as other Romanowsky stains. One of the commonly known Romanowsky stains is Leishman's stain, which is usually designed and used for air-dried peripheral smear. This study is designed to find out the effect of wet fixation and Pap stain on the air-dried peripheral smear by comparing it to Leishman's stain. Materials and Methods: The study included randomly selected 20 patients. With the informed consent, two peripheral smears were prepared with the blood samples. First set of samples were fixed with isopropyl alcohol and stained with Pap stain. Second set of smears were routinely air-dried and stained with Leishman's stain. Both the sets were analyzed for staining and morphological characteristics and statistically compared. Conclusion: After the study, it was quite evident that Pap stain does not give desired results on the peripheral smear, making it difficult for examination. Hence, Pap satin is not used for peripheral smear as it causes lysis of cells, change in cellular morphology, nondifferentiable among other cells, and many other complications for a cellular examination. Leishman's stain is the best commercially available stain for the peripheral smear examination.

Comparative evaluation of conventional cytology and a low-cost liquid-based cytology technique, EziPREP™, for cervicovaginal smear reporting: A split sample study.

Background:Liquid-based cytology (LBC) for cervical cancer screening offers several advantages over conventional cytology. However, the extremely high cost of the current approved devices precludes widespread application of LBC technique in resource-constrained countries. This study aimed to evaluate the performance of an indigenous low-cost LBC technique, EziPREP™ (EP), against conventional preparations (CPs) for cervical cancer screening.Materials and Methods:A cross-sectional split-sample study with consecutive cervical sampling was conducted on 515 women attending the clinic at our institute. CP smears were prepared as per the standard technique using spatula and endocervical brush followed by detaching the head of brush into the fixative vial of EP. The EP samples were processed as per the manufacturer's protocol. Both CP and EP smears were stained using standard Papanicolaou stain protocol. Both sets of smears were evaluated for staining quality, morphologic details, and cytologic diagnoses. Cytologic diagnoses were correlated with cervical biopsy findings, wherever available. Performance characteristics of the two techniques were calculated.Results:The unsatisfactory rate for CP was 1.0%, while on EP, 1.3% smears had inadequate cellular material. The staining quality and morphological details were comparable in both sets of smears. The detection of infections and epithelial cell abnormality was more, though not statistically significant in EP smears. There was a 98% concordance in cytologic diagnosis between CP and EP smears. Cytohistologic concordance was observed in 96% of cases for both CP and EP smears. Although the time taken for processing and staining of smears was higher for EP (2.5 min for EP per smear and 1.6 min for CP per smear), the screening time reduced from 6.5 min per smear for CP to 2.2 min in EP smears.Conclusion:EP provides monolayered cervical smears with vivid morphological details, leading to reduced screening time and relatively higher pick-up of infections and low-grade cervical lesions as compared to conventional smears. The availability of such low-cost devices may enable wider application of cytology-based cervical cancer screening in low-resource countries.

Rehydration of air dried oral smears using honey

Background: Papanicolaou stains, very reliable cytological staining in diagnosis of oral carcinoma. In routine practice, 95% ethanol is used to fix the slides while preparing for Pap smears. The problems associated with pap smear such as short of supply and storage of alcohol, wet fixing of slides and transporting them to the cytology centre if the Pap smears are prepared at a peripheral centre. If an alternative method of rehydrating the air-dried oral smears is known we can overcome these problems. Until the date only few studies regarding rehydration of oral smears as a substitute for wet fixation.Aim: The aim of this study is to use honey as an alternative for rehydrating air dried oral smears.Material and methods: This study was done over than 3 months. Two sets of oral smears wet and air dried ,Overall 40 smears were examined; The first set were fixed in 95% ethyl alcohol. The other set of smears were air dried and rehydrated with honey solution(2 parts honey and 1 part saline)Both the smears were stained with Pap stain, examined for preservation of cytomorphological features and evaluated by two experienced pathologists using the light microscope.Results and analysis: This study revealed that immersion of air dried PAP smears in honey for 30 minutes is effectively same as wet smear which is routine methodConclusion: Rehydration, followed by fixation of air-dried smears, is a simple, feasible, applicable and reliable fixation technique, which is comparable to the wet-fixed conventional technique, used for oral smears and can be applied for evaluation on a routine basis.

Human Palatal Neoplasm - A Cytopathological Approach

Background: Palatal neoplasm cases in human are very rare. More than 90% neoplasms of the palate is squamous cell type. Not only cytological pleomorphism but also nuclear anomalies are reported to be observed in oral squamous cell carcinoma (OSCC). But, site specific cytological pleomorphisms are not reported so far. Therefore, an attempt has undertaken to investigate the cytopathology of human palatal neoplasms, pattern of cervical lymph node (CLN) metastasis and to analyse the probable etiological risk factors associated with it in the present study. Methodology: In a hospital-based study, out of 136 oral cases, 9 palatal cases (6 male and 3 female) registered during May 2007 to May 2009 were included in this study. Detail case-history including the nature and types of addiction of each individual was recorded prior to the collection of samples. Two scraped exfoliated cytosmears were collected from the affected site on the pre cleaned-coded glass-slides. Collected cytosmears were immediately fixed in 1:3 aceto-alcohol (1 part of glacial acetic acid and 3 parts of ethyl alcohol). One set of smears was stained with Papanicolaou’s stain and the other was counter-stained with Giemsa’s stain for cytopathological analysis. American Joint Committee for Cancer Staging and End-Results Reporting (AJC) formulated TNM (Tumor-Node-Metastasis) System for staging of OSCC was followed. Result: Cytopathologically, a number of pleomorphic cytological atypias, such as, keratinized spindle cell (KSC), keratinized tadpole cell (KTC), keratinized strap (Antischkow) cell (KSC-A), large and small keratinized fiber cells (KFC), large and small keratinized round cells (KRC), micronucleated cell (MNC), plump keratinized squamous cell (PKSC) and non-keratinized malignant squamous cell (NMSC) were observed in such palatal neoplasms. Except NMSCs, all other cells were keratinized. Out of these, PKSC and MNC were well differentiated; KSC, KTC, KFC, KRC and KSCA were moderately differentiated and NMSC was poorly differentiated. Interestingly, in addition to the micronucleated cell (MNC), KSC-A was found to be a modal cytological atypia irrespective of age, site, sex and degree of pathogenicity which may be attributed to the addiction of tobacco in general and smoking in particular. Conclusion: Chewing and smoking of various form of tobacco and drinking of alcohol contribute a lot to the genesis of cytological pleomorphism in human palatal neoplasm. Typically atypical cells like KSC, KTC, KSC-A, KFC, KRC, PKSC, MNC and NMSCs although found less in number, modal occurrence of KSC-A along with the MNC in all the palatal neoplasm may be directly correlated with any form of tobacco smoking.

Risk factors for malaria in children presenting with fever or history of fever in rural Gambia

Successful control of malaria depends upon the detailed knowledge of its epidemiology, including the knowledge of environmental, behavioral, socio-economic and socio-demographic factors that influence its prevalence, and on the knowledge and use of preventive measures. We carried out a cross-sectional study to assess the risk factors for malaria in children presenting with fever in rural Gambia. Three hundred and seventy-six children aged 5 months to 10 years presenting to Farafenni health centre, the Gambia with fever or history of fever were enrolled. The socio-demographic characteristics, clinical findings, and knowledge and use of preventive measures in malaria were documented. Finger-prick blood sampling for malaria was obtained and two sets of smears (thick and thin) were prepared and stained with Giemsa stain. One hundred and eighty two children (48.4%) had malaria. The significant risk factors associated with malaria were non-use of insecticide treated bed nets [χ2 = 8.82, P = 0.01, (95% confidence interval 0.10–0.67)] and age [χ2 = 9.18, P = 0.004, (95% confidence interval 0.31–0.78)]. There were no significant associations identified between the development of malaria and parental education, occupation, knowledge of the cause of malaria, type of roof or wall in the home, or whether the parents reared livestock.

Comparison of the Routine Papanicolaou Staining Technique with the Rapid, Economic, Acetic Acid, Papanicolaou (REAP) Technique

Background : The universal stain for oral cytological screening for precancer and cancer of oral cavity is Papanicolaou stain which has been used in different laboratories with many modifications. Objectives : To assess the superiority of staining of smears by REAP technique compared to PAP technique. Material and Methods: 100 smears were collected from 50 patients. One set of smears were stained with conventional PAP technique and the other set with the REAP technique. In the REAP technique, the ethanol bath in the pre Orange G6, post Orange G6 and post EA50 stages is replaced by 1% acetic acid; Tap water is used instead of Scott's tap water and hematoxylin is pre heated in water bath to 60°C for rapid penetration. Methanol is used for final dehydration. The two sets of smears are observed by two independent observers and assessed for the optimal and suboptimal nuclear and cytoplasmic staining. The results are compared and analyzed statistically. Results : Good cytoplasmic transparency and optimal nuclear details were seen in REAP stained smears compared to the conventional PAP smears. The cost involved in REAP method was lesser compared to conventional PAP technique. REAP technique took 3 minutes for completion compared to PAP technique that involves a minimum of 20 minutes. Conclusion: REAP technique produces better stained smears that are cost effective and involve minimal time for mass screening of oral cancer as compared to conventional PAP smears.

Confocal microscopy and exfoliative cytology

Context:Early detection of potentially malignant lesions and invasive squamous-cell carcinoma in the oral cavity could be greatly improved through techniques that permit visualization of subtle cellular changes indicative of the neoplastic transformation process. One such technique is confocal microscopy. Combining rapidity with reliability, an innovative idea has been put forward using confocal microscope in exfoliative cytology.Aims:The main objective of this study was to assess confocal microscopy for cytological diagnosis and the results were compared with that of the standard PAP stain.Settings and Design:Confocal microscope, acridine orange (AO) stain, PAP (Papanicolaou) stain. The study was designed to assess confocal microscopy for cytological diagnosis. In the process, smears of patients with (clinically diagnosed and/or suspected) oral squamous cell carcinoma as well as those of controls (normal people) were stained with acridine orange and observed under confocal microscope. The results were compared with those of the standard PAP method.Materials and Methods:Samples of buccal mucosa smears from normal patients and squamous cell carcinoma patients were made, fixed in 100% alcohol, followed by AO staining. The corresponding set of smears was stained with PAP stain using rapid PAP stain kit. The results obtained were compared with those obtained with AO confocal microscopy.Results:The study had shown nuclear changes (malignant cells) in the smears of squamous cell carcinoma patients as increased intensity of fluorescence of the nucleus, when observed under confocal microscope. Acridine orange confocal microscopy showed good amount of sensitivity and specificity (93%) in identifying malignant cells in exfoliative cytological smears.Conclusion:Confocal microscopy was found to have good sensitivity in the identification of cancer (malignant) cells in exfoliative cytology, at par with the PAP method. The rapidity of processing and screening a specimen resulted in saving of time. It added a certain amount of objectivity to the process of arriving at a diagnosis.

Increased sensitivity of sputum microscopy with sodium hypochlorite concentration technique: A practical experience at RNTCP center

Background:In revised national tuberculosis control program (RNTCP), microscopic examination of sputum for AFB plays an important role in the initial diagnosis of tuberculosis. Bacillary concentration after decontamination and liquefaction by 5% sodium hypochlorite is useful in providing increased sensitivity and safety for handling of specimen.Materials and Methods:In this cross-sectional, prospective study, carried out at NKP Salve Institute of Medical Sciences and RC, Nagpur, patients were included according to RNTCP criteria. One set of smears was made according to the RNTCP guidelines while another set was prepared by concentration after decontamination with 5% sodium hypochlorite. Both set of smears were stained according to RNTCP method and were screened by two observers separately to remove observer’s bias and graded according to the RNTCP guidelines. A total of 591 sputum samples from 219 patients were included in the study with 168 males (76.71%) and 51 females (23.28%).Results:A total of 77 samples (13.02%) from 34 patients were positive by routine method whereas by concentration method 119 samples (20.13%) from 49 patients were found positive diagnosing 15 additional patients. This rise of 7.11% in sputum positivity over routine is highly significant (P=0.001021, χ2=10.78) with 44.11% increase in diagnosed cases.Conclusion:There is a statistically significant rise in smear positive cases after concentration with 5% sodium hypochlorite solution. Considering its low cost, decontaminating and liquefaction properties with better sensitivity, this method is safe and can be of vital importance; at least for smear negative cases.

Modified Papanicolaou staining protocol with minimum alcohol use: a cost‐cutting measure for resource‐limited settings

To devise a simple, cost-effective protocol for Papanicolaou (Pap) staining of cervicovaginal smears. Five hundred coded paired cervical smears were collected from women as part of routine cervical cancer screening. One set of smears was stained by conventional Pap staining protocol (CP) and the other by a modified protocol (MP) in which alcohol was replaced by 1% acetic acid in all the steps except during fixation and prior to mounting; in addition, one alcohol-based counterstain, OG, was omitted. The smears were examined blindly by the pathologists and then decoded. Each pair of smears was compared and the two protocols were analysed for staining quality and diagnoses by McNemar and chi-square tests. The staining quality in the MP was satisfactory. The nuclear and cytoplasmic features were comparable to the CP. Cytoplasmic transparency was maintained in the MP and the differential staining of blue/green and pink was acceptable to the pathologists and technicians. The diagnoses agreed in all cases and there was no compromise in interpreting the smears. With MP it took only 3-4 minutes to stain a batch of 50 slides. in contrast to the 20 minutes taken by CP. The MP used almost one-seventh of the amount of alcohol compared with CP, which translated into a significant cost reduction per smear. The improvised Pap staining protocol with minimum alcohol use is a simple, cost-effective and technician-friendly procedure that can be easily adopted in high-volume, resource-limited laboratories for mass cervical cancer screening.

Optimal fixation conditions for immunocytochemical analysis of estrogen receptor in cytologic specimens of breast carcinoma.

The techniques for immunostaining estrogen receptor (ER) in cytologic specimens have varied, as have the detection rates. The authors compared various fixation methods for their effect on ER detection in cytologic smears of breast carcinoma. Smears were prepared by gently scraping the cut surfaces of 47 resected breast carcinoma specimens and placing immediately in 1 of the following conditions: 1) a sequence of 10% formalin-methanol-acetone fixatives at -20 degrees C (Abbott method); 2) 10% formalin at room temperature; and 3) Carnoy's fixative at room temperature and then Papanicolaou stained (Carnoy's-Pap). Destaining of Carnoy's-Pap smears (Carnoy's-dPap) was initially attempted before ER staining. One set of smears was also air-dried for 3 minutes before using the Abbott method. Smears and corresponding tissue sections were immunostained with anti-ER antibody 6F11 using a similar protocol except for antigen retrieval, which was not initially applied on cytologic slides. All the ER-negative smears that had been fixed with 10% formalin or Carnoy's-Pap were restained after antigen retrieval. Agreement between cytologic and histologic findings was expressed by both concordance and the kappa coefficient. ER detection in smears processed with the Abbott method correlated best with findings from tissue samples, with an overall correlation of 91.5% (kappa = 0.80). Findings from air-dried smears were less optimal (concordance, 84.4%; kappa = 0.65), followed by Carnoy's-Pap (concordance, 71.4%; kappa = 0.45), formalin (concordance, 31%; kappa = 0.05), and Carnoy's-dPap (concordance, 29.4%; kappa = 0.04). Antigen retrieval converted most of the ER-negative smears to positive (18 of 20 smears in formalin and 6 of 8 smears in Carnoy's-Pap), leading to a final concordance of 93% and kappa = 0.83 for both conditions. Antigen retrieval also led to stronger staining intensity without causing false positivity. Antigen retrieval was found to greatly improve ER immunodetectability and staining intensity in formalin-fixed and Carnoy's-Pap smears. The former may offer an alternative to the Abbott method because of its easiness and the latter can be reliably used in archival Pap-stained smears for retrospective analysis of ER. Air-drying, destaining Pap smear, and fixation in formalin or Carnoy's-Pap without antigen retrieval are not recommended.

Utility of thick smears of bone marrow aspirate in pyrexia of unknown origin.

To determine the diagnostic efficacy of Giemsa stained thick smear of bone marrow aspirate for evidence of malaria in febrile individuals in whom the diagnosis was otherwise unknown. Descriptive case-series study. This study was conducted at the Department of Medicine, Sandeman Provincial Hospital, Quetta, in the Balochistan province of Pakistan, between January 1994 and December 1998. A total of 80 patients were included in the study, who presented with history of fever for more than 3 weeks with splenomegaly and pancytopenia. The bone marrow specimens of all these patients were examined. The bone marrow was aspirated from the posterior superior iliac spine. Three sets of smears were made. One set of thin smears was stained by Leishman's stain after air-drying and the second set of thin smears was air-dried, fixed with 97% methanol and later stained with 5% Giemsa stain. The third set of smears was a thick smear stained unfixed with 5% Giemsa stain. A temperature of 37 degrees C and a pH of 7.2 was maintained during the staining process. The presence of malarial parasite in the bone marrow was determined. Besides, the degree of megaloblastic change in erythroid series and the bone marrow cellularity were evaluated. The results obtained were recorded and statistically analyzed. Of the 80 bone marrows examined, 32(40%) showed the presence of malarial parasites, 19(23.75%) showed haematological malignancies, 16(20%) showed bone marrow suppression while 12(15%) showed megaloblastic changes. One (1.25%) case showed Leishman-Donovan bodies. Out of 32 cases showing the presence of malarial parasite in bone marrow, only 5(15.6%) revealed parasites, i.e. Plasmodium falciparum gametocytes on thin smear while 32(100%) cases showed Plasmodium falciparum trophozoites and gametocytes on thick smear. The ratio was 1:6.4. These 32 patients were divided into two groups, one group having mild anaemia (>8 g/dL) and the other severe anaemia (Hb<8 g/dL). In this study 20 patients had mild and 12 had severe anaemia. Megaloblastic changes were seen in 15 cases. Hypocellularity was seen in 3 patients with falciparum malaria while 28 showed normocellularity. The marrow was hypercellular in one patient. In patients presenting with prolonged fever, splenomegaly and pancytopenia, the Giemsa stained thick smear of bone marrow aspirates for evidence of malarial parasites was far superior to thin smear in diagnosing Plasmodium falciparum malaria.

Effect of circulation and discussion of cervical smears on agreement between laboratories

The study objective was assessing whether circulation of smears and discussion of those with differing interpretation can increase reproducibility between laboratories. The study included: the blind interpretation of a first set of 194 smears among seven laboratories, the discussion of smears with discrepant diagnoses during the previous phase and the blind interpretation of a second set of smears of same size and characteristics. After discussions, the overall weighted kappa increased in five laboratories (substantially in three : +50%, +27% and +20%). However, no change was observed in one laboratory and a slight decrease (-4%) in another. The latter interpreted the second set of smears at a longer time interval from discussions. Agreement improved for all diagnostic classes except low grade intraepithelial neoplasia (LSIL). Overall, the intervention increased diagnostic agreement, but its effect varied with laboratory and by diagnostic class and could be transient. Continued programmes of smear exchange and discussion appear to be advisable.

Interobserver Variability in Assessing Adequacy of the Squamous Component in Conventional Cervicovaginal Smears

We compared the interobserver reproducibility of estimating the adequacy of the squamous component of conventional Papanicolaou (Pap) smears using traditional and newly proposed criteria. Forty conventional Pap smears with varying degrees of squamous cellularity were reviewed by 13 observers who evaluated adequacy (satisfactory vs unsatisfactory) based on the traditional criterion of estimating 10% slide coverage. After being introduced to the new criterion and the reference images, the observers reevaluated adequacy on the same set of smears, using the new criterion and the reference images. With the original criterion of 10% slide coverage, 15 smears had a unanimous designation; the overall kappa value was 0.49 (P < .001). With the newly proposed adequacy criterion and reference images, 17 smears had a unanimous designation; the overall kappa value was 0.60 (P < .001). The difference in the kappa correlation coefficients was statistically significant (P = .007). While traditional and newly proposed criterion resulted in fair interobserver agreement, it seemed that the newly proposed criterion, along with the use of reference images, for evaluating adequacy of the squamous component of conventional Pap smears result in better interobserver reproducibility.

Estimating the sensitivity of cervical cytology: errors of interpretation and test limitations.

The objective of this study was to estimate: (i) the sensitivity of cytologists in recognizing abnormal smears; (ii) the sensitivity of cervical cytology as a method of detecting abnormal smears among those obtained in the presence of cervical intraepithelial neoplasia (CIN). Study subjects were 61 women with a histologically confirmed CIN identified through colpohistological and cytologic screening. For objective (i) new smears were taken from study subjects just before treatment, mixed with routine preparations, interpreted by unaware cytologists and then blindly reviewed by a group of three expert supervisors, who reached a consensus diagnosis. Cytologists classified as positive for squamous intraepithelial lesion (SIL) 30 of the 34 smears judged as positive by supervisors (100% of smears classified as high-grade and 67% of smears classified as low-grade SIL by the supervisors). Our approach, based on creating a set of smears with a high a priori probability of being positive, proved to be an efficient way of estimating errors of interpretation. For objective (ii), smears taken at the moment of diagnosis, just before biopsy, were also reviewed by the same supervisors. These CIN cases were identified among asymptomatic women independently of cytological findings and results are therefore not subject to verification bias. Among the 33 histological CINII/III, four (12%) smears had no atypical cells (three negatives and one unsatisfactory) at review. The same proportion was 26% (four negatives and one unsatisfactory) among the 19 histological CINI. No significant differences in smear content were found between the seven 'false negatives' and a sample of 'true positive' and 'true negatives' for a number of formal adequacy criteria (including presence of endocervical cells). Strong differences were found between positive smears taken just before biopsy and those taken just before treatment (in 11 women the first smear only was positive, while the opposite was never observed), suggesting an effect of punch biopsy in removing lesions.

Demonstration of Globi and Leprosy Bacilli by Fluorescence Microscopy.

Hagemann1 reported the superiority of a fluorescence method using berberine sulfate stain for demonstrating leprosy bacilli in nasal mucus and thick blood smears. The same year he2 proposed an auramin-fluorescence technic for the routine identification of acid-fast fast bacteria, especially tubercle bacilli. The suitability of this method for cultures of Myco. leprae has been confirmed by Kuster3 and for leproma smears by Kline and Leach.4Recently one of us (H.J.H.) has investigated the separation and concentration of acid-fast organisms from the tissues of leprosy patients. To cryochemed spleen† distilled water was added and direct impression smears were made from a piece of the tissue. One set of smears was stained with auramin O and examined according to the fluorescence procedure described by Richards and Miller.5A representative field is shown in the accompanying photomicrographs. The faint material in the background is non-acid-fast splenic cellular substance. The acid-fast bacilli have a characteri...