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1.
Introduction: A glacial acetic acid wash performed retrospectively or prospectively on visibly bloodstained cervical ThinPrep® specimens can reduce the unsatisfactory rate and increase the number of diagnostic epithelial cells. This study was undertaken to determine which specimens are most likely to benefit from a prospective glacial acetic acid wash. Methods: Bloodstained ThinPrep® specimens selected for routine lysing prior to processing were macroscopically assessed and scored based on the level of blood present (+ to +++). Both unlysed and lysed slides were prepared from each specimen and microscopically examined. Results: Fifty‐eight specimens (32 scored +, 12 ++ and 14+++) were assessed. Three unlysed slides prepared from the ++ specimens and 12 from +++ specimens were evaluated as unsatisfactory due to excessive blood and inadequate numbers of squamous cells. In contrast, only one of the unlysed slides from the 32 + specimens was considered to be unsatisfactory. Almost all the lysed slides were satisfactory and generally more cellular than the unlysed slides. Abnormal cells were found in four cases (both unlysed and lysed paired slides). Discussion: Although the acetic acid wash increases cellularity of bloodstained ThinPrep® specimens, prospectively lysing all bloodstained specimens is an unnecessary procedure. Lysing only very heavily bloodstained specimens prior to processing will reduce laboratory workload, costs and the possibility of specimen mix up. Occasionally a retrospective wash may be required but screening staff should be aware that although blood may be present on an unlysed ThinPrep® slide, a lysed slide should not be requested unless there are insufficient numbers of squamous cells present.  相似文献   

2.
OBJECTIVE: To characterize cervicovaginal cytology samples with < 5,000 squamous cells on the initial ThinPrep slide (Cytyc Corp., Boxborough, Massachusetts, U.S.A) and to attempt sample recovery using acetic acid. STUDY DESIGN: Cervicovaginal cytology samples with <5,000 squamous cells on the original ThinPrep slide and residuum were reprocessed by adding 3 mL of 3:1 CytoLyt (Cytyc)/glacial acetic acid with production of a second slide. Both slides were reviewed for squamous cell quantitation and the presence of background material and abnormal cells. RESULTS: From a total of 1,833 cases, 147 (8.0%) were identified for reprocessing; 71 (48.3%) were grossly bloody and 58 (39.4%) grossly cloudy. Reprocessing resulted in a second slide with > 5,000 squamous cells in 116 (78.9%) cases and was most effective on cloudy samples (89.7% recovery) and bloody samples (71.8% recovery). Abnormal cells were identified in 13 (8.9%) reprocessed samples. In all but 2 cases the abnormal cells were present on the initial slide and demonstrated the same degree of abnormality as the reprocessed slide but were fewer in number. CONCLUSION: Acetic acid recovery increases squamous cell recovery when initially inadequate, reducing the number of unsatisfactory cases and in rare cases identifying a cytologically significant lesion not apparent on the original slide.  相似文献   

3.
Objective:  To compare the accuracy of intraoperative fine needle aspiration cytology samples prepared by the ThinPrep method to conventional cytological methods. Specimen adequacy and turn around time (TAT) were also assessed.
Methods:  Fifty consecutive fresh tumours submitted for histological analysis were aspirated and each prepared as follows: (i) direct smear with H&E stain, (ii) direct smear with Pap stain, (iii) ThinPrep slide with H&E stain, and (iv) ThinPrep slide with Pap stain. The slides were randomly distributed to three cytopathologists for interpretation. The quality of the preparation, the diagnosis and the time needed for interpretation were recorded.
Results:  Accuracy was measured as the percentage of absolute agreement between the cytological and the histopathological diagnoses of the lesions. Histologically, there were 43 malignant and six benign lesions and one atypical lipoma. The TAT began when the slides/cytolyte specimens arrived at the lab and ended with the pathologist's diagnosis.
Conclusions:  In terms of accuracy and specimen adequacy, ThinPrep slides with Pap stain is the best procedure. This advantage however is offset by the longer testing time.  相似文献   

4.
OBJECTIVE: To examine cells that were retained on sampling devices used to collect ThinPrep (Cytyc Corp., Boxborough, Massachusetts, U.S.A) Pap smears in order to evaluate both the number and significance of cells that are routinely discarded with these devices after liquid-based specimens are collected. STUDY DESIGN: One hundred Pap smears from 100 women were prospectively procured after gynecologic Pap smears were collected for the ThinPrep Pap test. The sampling end of the collection devices was cut off and placed in a vial that contained SUREPATH preservative fluid (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A). The residual cell samples were processed using the SurePath PREPSTAIN slide processor (TriPath). A single liquid-based slide was prepared from the sampling devices from each of the 100 specimens collected. The slides produced from the discarded devices were reviewed for the following: squamous cells, endocervical component, epithelial cell abnormalities and miscellaneous findings. The slides prepared from the "throw-away" (TA) material were subsequently compared with the primary ThinPrep Pap smear slide. RESULTS: Twenty-five percent of the TA samples had an equal or greater number of squamous cells per high-power microscopic field when compared to the primary ThinPrep slide, with 8% of the TA slides demonstrating greater overall cellularity. An endocervical component was present on 27 of 66 cervical samples (40.9%). Three of five cases (60%) interpreted as atypical squamous cells of undetermined significance had similar cells on the TA slides. Two cases of atypical glandular cells of undetermined significance had no abnormal cells on the TA slides. Twelve of 14 cases (85.71%) of low grade squamous intraepithelial lesion contained similar cells on the TA slides. Two of four cases (50%) of high grade squamous intraepithelial lesion also had similar abnormal cells on the TA slides. Miscellaneous findings included 1 case of benign endometrial cells and 4 Candida infections present on both preparations, along with 1 case of Trichomonas vaginalis organisms present on the ThinPrep slide only. In 1 specimen, several multinucleated histiocytic giant cells were present only on the TA slide. CONCLUSIONS: Specimens prepared from TA collecting devices used for the ThinPrep Pap test are less sensitive than the primary specimen for the detection of cervical lesions. This is in contrast to split-sample studies involving ThinPrep and conventional smears. Our study documented the presence of normal and abnormal cells discarded from ThinPrep sampling devices in a high percentage of cases. Discarded abnormal cells on the TA slides were, however, few when compared to the primary specimen, with only 1 exception involving a high grade lesion.  相似文献   

5.
Objective:  Large loop excision of the transformation zone (LLETZ) has become standard of care in the management of cervical squamous neoplasia and with cone biopsy glandular intraepithelial neoplasia. Controversy remains about the long-term effects of this traumatic procedure. The aim of this study was to count and compare the number of endocervical glandular cell groups in pre- and post-LLETZ cervical preparations using liquid-based cytology to establish a cyto-morphological correlate of destruction of the transformation zone.
Methods:  The cytology/histology correlation audit records of the Cytopathology Department of St Luke's Hospital in 2003 and early 2004 were used to select patients with a cytological diagnosis of high grade dyskaryosis followed by LLETZ. Only those cases with post-LLETZ cytological follow-up were selected. Cases using conventional smears were excluded. One hundred and twenty slides (60 pairs of slides) in total were retrieved. The cases underwent review and all groups of >3 glandular cells in each slide were counted by AM while blinded as to whether smears were pre- or post-LLETZ. Medians were compared using a Mann–Whitney U -test.
Results:  The median number of groups of endocervical glandular cells of the pre-treatment group was 5.5 and of the post-treatment group was 2.0. There were significantly fewer endocervical glandular cell groups in the post-LLETZ population ( P  = 0.03).
Conclusions:  The number of endocervical glandular groups in cervical cytological preparations decreases significantly following LLETZ procedure. This suggests that cytological follow-up may not be as useful in glandular neoplasia cases. Few or absent glandular cell groups in post-LLETZ preparations may have implications for adequacy assessment.  相似文献   

6.
Objective:  To define a minimum acceptable total squamous cellularity for (ThinPrep®) liquid-based cervical cytology (LBC) specimens using quality control techniques.
Methods:  Two hundred LBC preparations were made containing varying numbers (<200) of severely dyskaryotic squamous cells and with varying total cellularities.
Results:  Ninety-eight per cent of the LBC preparations that were missed by one or more of three cytoscreeners had fewer than 16 abnormal objects (single dyskaryotic cells or clumps of cells) and 87 dyskaryotic cells. The minimum ratio of dyskaryotic to total squamous cells that, in a preparation of 5000 squamous cells has a probability of at least 0.98 that 87 or more dyskaryotic cells will be present is 1 : 47. Twenty-three preparations diagnosed as abnormal had ratios of dyskaryotic to total squamous cells of between 1 : 2.5 and 1 : 4596. There is thus no feasible minimum acceptable squamous cellularity that will give an acceptable probability of detection of all specimen vials containing abnormal cells in the observed proportions.
Conclusions:  It is suggested that the minimum acceptable cellularity for LBC specimens is set pragmatically by the screening programme to give a feasible percentage of repeat tests.  相似文献   

7.
Basic Blue 148: A Rapid Stain for T Helper Cells   总被引:1,自引:0,他引:1  
After brief exposure to an aqueous solution of the oxazine textile dye C. I. basic blue 148 following fixation in 37% formalin, 95% ethanol and glacial acetic acid, T helper cell nuclei and cytoplasm in specimens of peripheral blood displayed a deep red-violet color. No other cell in normal blood or bone marrow specimens showed intense staining of this type. The total staining time is 1 min. Basic blue 148 stain is a promising technique for hematology and immunology laboratories as a rapid screening test for T helper cells in blood specimens using a microscopic slide and ordinary incandescent illumination.  相似文献   

8.
Introduction:  This poster aims to provide a discussion point for the calculation of screener performance. LBC has brought about changes in the way slides are interpreted, single dispersed isolated dyskaryotic cells take on a new meaning and the process of quality control, rapid review has changed. These changes challenge the rationale behind screener sensitivity calculations especially as many laboratories are in an early learning phase with regard to LBC.
Method:  Screener sensitivities and the PPV of reporting consultants for a period of six months post LBC conversion are compared with those since the introduction of LBC.
Results:  Screener sensitivities have dropped below the 95% threshold for high-grade dyskaryosis.
Discussion:  The change in rapid review or preview from a partial stepped rescreening of a conventional smear to a full rescreening of LBC slides has meant that all missed abnormalities that may not have been visualised in the conventional slide have a greater possibility of detection in the LBC slide. In analysing screener sensitivity a holistic approach that assesses the reasons for missing or misdiagnosing high-grade abnormalities is advised. Over reporting by consultants as indicated by PPV and slide review should be taken into account when there is a suspected poor performer. The recent move to refer all mild dyskaryotic smears for colposcopic assessment and the EQA requirement for screeners to detect dyskaryosis without the necessity for grading suggests that there may be a need to reassess the basis of current screener sensitivity calculations.  相似文献   

9.
Objectives:  To determine the degree of interobserver variation in the assessment of conventional cervical smear adequacy as defined by The Bethesda System (TBS) 2001, and to determine the effect of using reference images of known squamous cellularity when performing squamous adequacy assessments.
Methods:  Experimental pre-test/post-test design utilizing 70 conventionally prepared cervical smears. Sample smears containing scant squamous cellularity were independently rated on two occasions by six cytotechnologists. Time 1 was without the use of reference images, and Time 2 was aided by cellularity reference images. The κ statistic was used to compare rater agreement.
Results:  The level of agreement increased from an average κ of 0.26 (SD 0.10) for Time 1, to an average κ of 0.40 (SD 0.15) for Time 2. The difference in mean κ values at the two assessments was statistically significant ( t  = 3.71; P  = 0.002). Unanimous agreement among the raters was observed for 15 samples (21.42%) at Time 1 (only one of which was classified as unsatisfactory) and 21 samples (30.00%) at Time 2 (12 of which were classified as unsatisfactory).
Conclusion:  Interobserver agreement increased after cellularity reference images were implemented. Using TBS 2001 squamous adequacy criteria and images of known squamous cellularity as references resulted in a decreased number of smears reported as satisfactory.  相似文献   

10.
Objective:  To describe the cytological aspect of peritoneal washings in benign multicystic peritoneal mesothelioma (BMPM).
Methods:  Three peritoneal washing specimens stained by standard cytological and histological procedures and analysed by light microscopy.
Results:  The specimens showed an abundance of monomorphous mesothelial cells devoid of atypia or mitoses. The mesothelial cells were calretinin positive. They also showed numerous squamous metaplastic cells arranged in flat sheets or isolated cells. The background contained some inflammatory cells.
Conclusion:  The combination of cytology of the peritoneal washing, histology (cell block and surgical specimen) and clinical history allow differentiation of BMPM from other cystic lesions (cystic lymphangioma and malignant mesothelioma).  相似文献   

11.
Objective:  BK virus (BKV) may be associated with interstitial nephritis in renal transplant recipients and this can lead to irreversible chronic allograft dysfunction. Early diagnosis of BKV nephropathy determines its progress because no specific antiviral therapy exists. Urine cytology, detection of viral DNA in urine or blood and renal biopsy are the main diagnostic tools. The purpose of this study was to evaluate the use of urine cytology for diagnosis of BKV replication in renal graft recipients.
Patients and methods:  We studied 32 de novo renal transplant recipients prospectively with sequential urine samples for a period of 1 year. Thin-Prep methodology was used to prepare the slides. Cytology results were correlated with polymerase chain reaction (PCR) in urine and blood.
Results:  Decoy cells indicative of BKV infection were detected in 14 (7.3%) of the 190 urine samples derived from 11 recipients. In three cases with positive decoy cells, BK viraemia and viruria were simultaneously identified. In a further three cases, BKV active replication was confirmed in urine by both cytology and PCR.
Conclusions:  Urine cytology is an easy and rapid method of detecting decoy cells in cases where renal biopsy is not possible. However, the low incidence of detection of decoy cells in the present study, together with poor correlation with PCR results, questions its sensitivity and specificity in diagnosing BKV reactivation.  相似文献   

12.
We studied the characteristics of cytoplasmic microtubule reassembly from endogenous tubulin pools in situ using a Brij 58-lysed 3T3 cell system. Cells that were pretreated in vivo with colcemid retain endogenous tubulin in the depolymerized state after lysis. When lysed cells were removed from colcemid block and incubated in GTP-PIPES reassembly buffer at pH 6.9, microtubules repolymerized randomly throughout the cytoplasm, appeared to be free-ended and were generally not associated with the centrosomes. However, tubulin could be induced to polymerize in an organized manner from the centrosomes by increasing the pH to 7.6 in the presence of ATP and cAMP. Microtubules polymerized in ATP had significantly longer lengths than those assembled in GTP or UTP. When cells not treated with colcemid were lysed, the integrity of the cytoplasmic microtubule complex (CMTC) was maintained during subsequent incubation in reassembly buffer. However, in contrast to unlysed, living cells, microtubules of lysed cells were stable to colchicine. A significant fraction of the CMTC was stable to cold- induced disassembly whereas microtubules reassembled after lysis were extremely cold-sensitive. When cells not treated with colcemid were lysed and incubated in millimolar Ca++, microtubules depolymerized from their distal ends and a much reduced CMTC was observed. Ca++ reversal with EGTA rapidly resulted in a reformation of the CMTC apparently by elongation of Ca++ resistant microtubules.  相似文献   

13.
Objectives:  To assess the extent of lubricant use by smear-takers and the effect of lubricant contamination of ThinPrep® processed cervical cytology samples.
Methods:  All primary care smear-takers were sent a questionnaire on lubricant type and frequency of use. Fifty cervical cytology samples were then contaminated with incremental amounts of K-Y® jelly, 50 samples contaminated with incremental amounts of Aquagel® and ten non-contaminated vials were processed using the ThinPrep® T2000 processor followed by Papanicolaou staining. The morphological appearances of lubricant contamination were described microscopically and formal cell counts performed on all slides.
Results:  Seventy of 94 (74.5%) primary care smear-takers indicated lubricant use of whom 9/70 (12.8%) used Aquagel® and 61/70 (87.2%) used K-Y® jelly. K-Y® jelly appeared as mucoid blue deposits in the slide background whereas Aquagel® appeared as pink stringy background material. Cell counting showed a significant difference between Aquagel® and K-Y® jelly contaminated slides compared to the original non-contaminated preparations for all fields and the average fields ( P  < 0.001) with a significantly higher count for the original non-contaminated slides than the lubricant contaminated groups.
Conclusion:  Lubricant contamination of ThinPrep® cervical cytology samples may result in reduced cellularity of the subsequent slide. This study provides evidence-based data to support British Society for Clinical Cytology recommendations for no lubricant use when taking cervical samples.  相似文献   

14.
A. G. Siebers, J. A. W. M. van der Laak, R. Huberts‐Manders, J. E. M. Vedder and J. Bulten Accurate assessment of cell density in low cellular liquid‐based cervical cytology Objective: Scant cellularity is the most important source of unsatisfactory liquid‐based cytology. Although still being debated, low cellularity is thought to compromise the detection of squamous lesions. Thus, reliable assessment of cellularity is essential. The aim of the present study was to determine the cellularity range for ThinPrep® slides of low cellularity and to establish the most accurate cell‐counting protocol. Methods: A series of 60 ThinPrep cases representing the full spectrum of adequate, ‘satisfactory but limited by’ (SBLB) and unsatisfactory reports were included. Two cell‐counting protocols with three different magnifications, using ×10, ×20 and ×40 objectives, were evaluated and related to the true cellularity, together with a reassessment of the degree of adequacy originally reported. The cell‐counting protocol that showed the highest correlation coefficient was considered the most accurate. Results: Based on seven (re)assessments a majority score for adequacy was established. There were 42 cases with a majority score ‘unsatisfactory’ or ‘SBLB’ (low cellularity) of which 41 contained fewer than 20 000 squamous cells; and 18 cases with a majority score ‘satisfactory’ of which one had fewer than 20 000 cells. The cell‐counting protocol that showed the significantly highest correlation with the reference standard was the Stichting Kwaliteitsbewaking Medische Laboratoriumdiagnostiek (SKML) protocol with a ×10 objective. Conclusions: ThinPrep slides reported as unsatisfactory or SBLB were shown to contain fewer than 20 000 squamous cells. The most accurate protocol for estimating the cellularity of these slides was cell counting in five non‐adjacent microscope fields along the horizontal axis and five along the vertical axis of the slide with a ×10 objective and applying a correction factor of 1.24× to correct for underestimation of the true cellularity.  相似文献   

15.
16.
Smear the pollen mother cells of a single anther from each flower bud on a clean dry slide, using a small scalpel. Flood the slide with Belling's acetocarmin and heat for a second over an alcohol flame. Examine under the microscope to determine the stage of microsporogenesis. If the stage is satisfactory, smear the remaining anthers in the same manner, but fix and stain them by immediate immersion, face downward, in a petri dish full of hot (steaming) acetocarmin for from 1 to 10 minutes. Then rapidly transfer thru the following mixtures: two parts 99% (glacial) acetic acid plus one part absolute ethyl alcohol; one part acetic acid plus two parts absolute alcohol; and finally one part acetic acid plus nine parts absolute alcohol. The slides are then to be dehydrated completely by 1 to 2 minutes immersion in pure absolute alcohol, and cleared 2 to 3 minutes in a mixture of xylene and absolute alcohol in equal parts. The preparations are then made permanent by mounting each with balsam and a cover glass. The whole process takes from 5 to 15 minutes and is particularly recommended for chromosome counts.  相似文献   

17.
当今的蛋白质组学研究,都是先裂解细胞放出蛋白质,然后对蛋白质溶液进行各种分析.对于红细胞来说,它的裂解产物也称"溶血液",其中主要成分有血红蛋白A1,A2,A3和碳酸酐酶(CA)等.本实验室用未裂解的完整的活体红细胞直接进行电泳,观察其释放出来的血红蛋白(hemoglobin,Hb),建立了淀粉-琼脂糖混合凝胶中红细胞的电泳释放实验.电泳释放可分为"初释放"(一次通电完成电泳,此时有Hb释放出来)和"再释放"(电泳过程中断电-再通电,又有Hb释放出来).本实验室在"初释放"实验中发现了"HbA2现象",并通过Hb交叉电泳发现了HbA2与HbA1的相互作用;利用初释放型双向对角线电泳发现红细胞内HbA2与HbA1结合存在;对电泳释放出来的"HbA2现象"成分做SDS-PAGE及质谱分析,发现Prx-2(Peroxiredoxin-2)可能参与"HbA2现象"的形成;在研究"再释放"实验中发现了"Hb多带再释放现象",在此基础上创建等渗再释放、低渗再释放、等低渗全程再释放及再释放型双向对角线电泳;两种红细胞(全血中的红细胞和由它分离出来的游离红细胞)再释放的比较研究;血浆成分对红细胞再释放的影响等.以上研究方法的建立为活体细胞内蛋白质存在状态的研究提供了基础,并开辟了新的研究途径和领域.  相似文献   

18.
Objectives:  Degenerative change caused by delay in processing contributes to false-negative and false-positive diagnosis of urothelial carcinoma in cytology. The aim of the study was to see if the use of a collection fluid for urine samples made a significant difference to urine cytology diagnosis, and if one was better suited for routine use in the hospital laboratory. Three cell collection fluids were evaluated by analysing the preservation and degeneration of cells in urine samples, as was the routine preparation which did not use a collection fluid.
Methods:  In the design study 50 voided urine specimens were taken at random from the hospital haematuria clinic. Three commercially available collection fluids cytolytTM, cytospin® and cytoRich®Blue and the hospital's routine conventional preparation of urine were compared. The degree of degeneration, and so preservation, was assessed by a table of chosen criteria; then ranked and analysed by Friedman's nonparametric test, at P  = 0.05. A second table showing the cell content of each slide was also made.
Results:  These showed no significant diagnostic difference between the collection fluids, but there was a significant difference between the collection fluids and the routine preparation. Minor differences that do not affect diagnosis, such as crystals and ghost red blood cells, were noted in cytospin® and cytoRich®Blue.
Conclusion:  It is recommended that a collection fluid is used. This choice should be made after health and safety issues and cost are considered.  相似文献   

19.
OBJECTIVE: To evaluate the feasibility of performing multicolor interphase fluorescence in situ hybridization (FISH) on ThinPrep slides of transitional cell carcinoma (TCC). STUDY DESIGN: Slides from 20 voided urine specimens were prepared by the ThinPrep technique (Cytyc, Boxborough, Massachusetts, U.S.A.), pretreated using a pretreatment kit and subjected to hybridization with the multicolor FISH probe UroVysion (Vysis, Downers Grove, Illinois, U.S.A.). Archival slides were placed in xylene, destained in alcohol and washed prior to pretreatment. Urines from patients with cytology-positive, biopsy-proven grade 1 (n = 5), 2 (n = 7) and 3 (n = 5) TCC and negative cytology and biopsy (n = 3) were selected. Freshly prepared (n = 10) and archival (n = 10) slides were used. RESULTS: All carcinoma cases were FISH positive (> 5 cells with complex abnormalities of > or = 2 studied chromosomes per slide). None of the normal samples were aneusomic. Gain of chromosomes 3, 7 and 17 constituted the majority of positive cases. Proper destaining and slight decrease in stringency wash conditions enabled reliable detection of signals in archival cases. CONCLUSION: Routine ThinPrep slides can be used for multicolor interphase FISH analysis of urine cytology specimens. Archival slides provide the opportunity to analyze by FISH the nature of atypical cells identified by cytology. This revised method allows FISH technology more accessibility for routine use in cytology laboratories.  相似文献   

20.
Objectives:  Ideally, head and neck aspiration should be performed by trained aspirators within the setting of a one-stop clinic, where smeared material is available for immediate assessment. However, this may not always be possible for practical reasons and the use of liquid-based techniques in head and neck cytology is increasing. Although liquid-based cytology has been extensively validated for use in gynaecological cytology, no studies have investigated whether or not a single ThinPrep ® slide is representative for head and neck aspirate specimens. We performed a prospective audit of head and neck fine needle aspiration specimens processed by the ThinPrep ® method to investigate whether a single ThinPrep ® slide was representative.
Methods:  A prospective audit of 115 consecutive head and neck aspirates was carried out. A single ThinPrep ® slide was prepared and a diagnosis recorded. The remainder of the specimen was then spun down and prepared as a cell block. The ThinPrep ® and cell block diagnoses were compared.
Results:  In 36 cases (31%), the cell block provided additional information that contributed to the diagnosis. In 14 (12%), the cell block was regarded as essential to the diagnosis.
Conclusions:  A single ThinPrep® slide may not provide representative diagnostic material in all head and neck aspirates. This should be taken into consideration when contemplating the use of liquid-based methods for non-gynaecological cytology.  相似文献   

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