Cytoplasmic Details Research Articles

Étude de l’item d’interférence dans un dossier de validation de méthode : cas de la coloration HPS après fixation en formol-zinc et comparaison à la fixation en formol tamponné à 4 %

The department of neuropathology of Sainte-Anne Hospital uses zinc-formalin as the fixative agent for its samples. No publication referenced in Pubmed has proven the validity of this fixative agent. In the context of the accreditation of our standard staining (HPS for Hemalun-Phloxin-Saffron), we started a file for the validation of this method in which the fixative agent constitutes an «interfering» substance which can modify the quality of the technique. The aim of this study was to prove that the use of zinc-formalin as a fixative agent is as suitable as the fixation with 4% buffered formalin. A cohort of samples fixed by zinc-formalin and by 4% buffered formalin was performed on fresh samples, then cut and stained by HPS. The slides were interpreted by three pathologists (one of them was outside our centre) ``blind'' to the fixative agent and they evaluated four criteria (general quality of the staining, components of the extracellular matrix, cytoplasmic details, and nuclear details) and scored them (from 0to 3) according to the Association française en assurance qualité (AFAQAP) recommendations. The cohort included 43samples. The results of the analysis showed that for samples fixed by zinc-formalin, three of the four criteria obtained significantly a better score than the samples fixed by classical formalin. Our results show that the zinc-formalin fixative does not constitute an ``interfering'' agent for the quality of the HPS staining for neuropathological samples.

Fine needle aspiration cytology of cervical lymph nodes: Comparison of liquid based cytology (SurePath) and conventional preparation.

Fine needle aspiration cytology (FNAC) is the first diagnostic step in patient with cervical lymphadenopathy because of its simplicity, safety and early availability of the results. Liquid-based cytology (LBC) is an alternative processing method which is used for both gynecological and nongynecological samples. Literature reviewed show few studies comparing LBC with conventional preparation (CP). The present study was undertaken to evaluate the efficacy of LBC and comparison of LBC and CP in cervical lymphadenopathy. In this prospective study, a total of 75 cases of FNAC with cervical lymphadenopathy were included. The first pass was used for CP followed by LBC with the use of SurePath (SP) technique. Both the smears were compared for cellularity, background containing blood, cell debris, lymphoglandular bodies, stromal fragments, cytoarchitectural pattern, etc., by semiquantitative scoring system. There was no statistical difference in the cellularity, cell architecture, and monolayer cells (P > .05). On the basis of background containing blood, cell debris, lympho-glandular bodies, stromal fragments (P < .001), nuclear, and cytoplasmic details (P < .05), LBC was found to be superior to CP. LBC is a relatively simple technique and superior to CP in respect of better nuclear and cytoplasmic details with loss of background blood and debris. It has a diagnostic accuracy equivalent to that of CP. However, use of both LBC and CP can result in better diagnostic accuracy.

A study on qualitative comparison between cryostat and conventional technique for histopathology diagnosis

Introduction: Intraoperative consultation by frozen section technique is an invaluable tool for immediate diagnosis. The correlation of intraoperative frozen section diagnosis with final diagnosis on permanent section is an integral part of quality assurance in surgical pathology. Its accuracy and limitations vary with different anatomical sites. Aims &amp; Objective: Qualitative morphological comparison between frozen section and routine formalin fixed paraffin embedded sections in different tissues, to assess the accuracy of frozen section, to detect the number and type of discrepancies and to assess the causes for discrepancies. Material and Method: The present study retrospectively reviewed frozen sections performed in the pathology department, Sir-t hospital, government medical college, Bhavnagar during a period of 2 year. Diagnostic accuracy of frozen section and its morphological quality and reliability in comparison to histopathology was evaluated by 2 pathologists in a blinded fashion for the following parameters: cellular outline, nuclear and cytoplasmic features, staining pattern and overall morphology. The results were compared to the routine formalin fixed paraffin sections to evaluate diagnostic accuracy. Discordant cases were reassessed to find the reasons for discrepancy. Result: Diagnostic accuracy of frozen section was 95.1% with false negative case are 4.8% and no false positive case. Statistical analysis showed that most common indications of frozen section in the present study was presence/typing of neoplasm (61.2%) followed by assessment of margin (35.6%) and assessment of nodal status (3.2%). It was observed that nuclear details, cellular outline and overall morphological quality of frozen section was slightly inferior to that of routine histopathology section, however, staining and cytoplasmic details were comparable. The discrepancies were mainly due to technical artefacts (9.7%), sampling error (3.2%) interpretation error (1.6%), and partly due to lack of interdepartmental communication. Conclusion: Frozen section diagnosis is very useful and highly accurate procedure. Gross inspection, sampling by pathologist, frozen complemented with cytological and histological review and intimal cooperation with surgeon can avoid certain limitations and provide rapid, reliable, cost effective information necessary for optimum patient care.

Cytomorphologic analysis of wet fixed and air dried oral buccal smears rehydrated with normal saline: A comparative study

Introduction: Cytology is useful in disease diagnosis. The science of cytopathology is currently well standardized with two major branches, exfoliative and aspiration biopsy. The limitations of cytological diagnosis, are shrinking day by day. Exfoliative cytology is the study of cells exfoliated from body cavities. These exfoliated cells, from oral buccal mucosa are analysed by making smears. Smears are immediately fixed in alcohol for a period of 20 minutes and then stained with Pap stain. H & E, and Pap stains, are generally regarded as the best stains for assessment of chromatin pattern in cytologic smears since they ensure maximum resemblance with the corresponding cells in tissue sections. Cytological smears are used nowadays for mass screening purposes in camps. Taking bottles of alcohol for fixing and ensuring to store them in tightly sealed copulin jars, with smears, may be difficult. Air-dried smears may be used to overcome such practical difficulties. Rehydration of these air-dried smears can be a good alternative to wet fixation. Methodology: 20 healthy subjects were randomly selected and 2 oral buccal smears were taken from the same subject. One was fixed by routine alcohol wet fixation, the other was air dried for 24 hours, then rehydrated with saline for 20 minutes and fixed with alcohol. The slides were assessed after labelling. The scoring criteria was based on processing, preservation, nuclear details, cytoplasmic details and background staining. Two observers, blinded to the method of fixation, scored the slides, to avoid any bias. Results: The air-dried smears rehydrated with saline were almost equal, or even superior, in few characteristics, to wet fixed smears. The ease of processing was equal in both, while the nuclear details and cytoplasmic details were superior in saline rehydrated smears. However, the background stain was slightly darker in saline rehydrated smears compared to wet fixed smears. A student paired t-test was also done (p=0.639).

Comparison of Five Cytochemical Stains used for Fine Needle Aspiration Cytology of Breast and Lymph Node Lesions: A Comparative Study

Introduction: Fine Needle Aspiration Cytology (FNAC) is an established diagnostic tool aiming at an early diagnosis of mass lesions which in turn aids in planning further management of the patients. The need for minimal turnaround time with good quality of cell morphology has encouraged the development of different staining techniques. Quality Index (QI) of cytochemical stains is evaluated with a scoring system based on morphological features. Aim: To find rapid staining methods which were economical without compromising on the quality of FNAC smears. QI of five stains namely Papanicolaou stain (Pap), Haematoxylin and Eosin (H&amp;E), May Grunwald Giemsa (MGG), Modified Ultrafast Papanicolaou (MUFP) and Rapid Economic Acetic acid Pap stain (REAP) were studied and compared. Materials and Methods: This was a prospective study conducted in the Department of Pathology, Kempegowda Institute of Medical Sciences, Bengaluru, Karnataka, India, from August 2011 to July 2013. FNAC of 80 mass lesions including 44 of breast lesions and 36 lymph nodes were included in this study. A minimum of five smears were made in each case and stained with Pap, H&amp;E, MGG, MUFP and REAP. The QI of stain was assessed by the scores given for background of the smear, overall staining, and preservation of cell morphology, nuclear characteristics, cytoplasmic details and air drying artifacts. Statistical analysis of the data was done using chi-square test and Statistical Package for the Social Sciences (SPSS) version 15.0. Results: Pap stain showed maximum QI score followed by H&amp;E, REAP, MGG and MUFP in case of breast lesions. In cases of lymph node highest QI was again with Pap followed by H&amp;E, REAP, MUFP and MGG. Good cytomorphology with lesser expenditure and turnaround time was obtained by REAP and MUFP stains. Conclusion: Pap stain is still the best for cell morphology whereas MUFP and REAP are good, economical and with lesser turnaround time and can be used in routine cytopathology.

Diagnostic utility of cell blocks and an immunomarker panel in the cytological evaluation of serous effusions

Introduction: Cell blocking (CB) has gained wide acceptance as an integral diagnostic tool in cyto-diagnosis. It allows retrieval and concentration of exfoliated cells in fluid specimens to form paraffin blocks that are interpreted in a biopsy like fashion. The aim of this study was to compare the diagnostic utility of CB to conventional smears (CSs) in the evaluation of serous effusions and to evaluate whether the tested immunomarker panel increases the diagnostic yield of malignancy. Materials and Methods: Serous effusion specimens were processed using both techniques; CSs and CB. An immunomarker panel (Ber-EP4, EMA E29, D2-40 and desmin) was then applied on CB sections. Results: CB enhanced the cellularity; it increased the diagnostic yield by 27.4% over CSs. Both nuclear and cytoplasmic details were better demonstrated in CBs. CBs were superior in the preservation of architectural patterns as acini, papillae and sheets. The diagnosis of CBs concurred with that of CSs in 79% of cases, however the diagnostic yield of malignancy increased by 9.6% using CBs. Adding the immunomarker panel totally eliminated the suspicious category and further increased the diagnostic yield for malignancy by 9.7% over CBs and by 19.3% over CSs. Furthermore, categorization of malignant cases into primary malignant mesothelioma and metastatic carcinoma was feasible after thorough interpretation of the combined immunoprofile. Conclusion: CB represents a cost-effective adjunct to CSs in the evaluation of serous effusions. It increases the cellular yield, provides superior cytomorphology, architectural preservation, and supplies material for ancillary studies. Keywords: Serous effusions, Cell blocks, Immunohistochemistry, Marker panel.

The Diagnostic Utility of Cell Block Preparation with Conventional Cytological SmearsA Cross Sectional Study

Background: Cytological examination of serous fluids aspirated is a simple and relatively non-invasive technique to diagnose whether the effusion is malignant or benign. Cell block preparation along with conventional smear increases the sensitivity of detecting malignancies, and also has the ability to reduce false-positive interpretations. Methods: A total 68 samples of body fluid (pleural and ascitic) specimens were examined for conventional cytological smear (CS) and cell block method (CB) over a period of one year. Out of 68 fluids, 40 were pleural fluid and 28 were ascitic fluid. Each fluid specimen was examined by conventional smear technique as well as cell block technique. The morphological details, cellularity, architecture, nuclear and cytoplasmic details were studied in both CS and CB techniques. Result: A total 82.35% smears had adequate material; while of the total cell blocks, 75% cell blocks had adequate material. A total of 11.76% cases were malignant on smears,5.88% were suspicious of malignancy, 64.7% were benign/non-neoplastic lesions. A total 13.2% cases were malignant on cell block, 1.47% were suspicious of malignancy, 60.29% were benign/non-neoplastic lesions. Sensitivity, positive and negative predictive value and accuracy of cell block technique were greater than that of FNAC smears Conclusion: For the final cytodiagnosis of body fluid, there is statistically significant difference between the two techniques. Cell blocks prepared from the residual fluid specimen can be useful for more definitive diagnosis, with advantage of IHC and special stains where required.

Comparison of Conventional Cytology, Liquid-Based Cytology, and Cell Block in the Evaluation of Peritoneal Fluid in Gynecology Malignancies

Background: Peritoneal washing cytology has become an accepted method in evaluating gynecology malignancies. Objective: The aim of this work was to compare the conventional cytology, liquid-based cytology (LBC), and cell block in the evaluation of peritoneal fluid. Materials and Methods: A total of 48 cases with ovarian and uterine malignancy were included in this study. The age of the patients varied from 15 to 71 years. All the 48 cases were subjected to conventional smear, LBC, and cell block preparation. Results: The results of LBC and conventional methods were in line with all cytological criteria, except for the background (p = 0.045), but no significant difference was found regarding adequacy (p = 0.12), cellularity (p = 0.13), cell architecture (p = 0.751), nuclear details (p = 0.96), and cytoplasmic details (p = 0.32). The kappa correlation between conventional cytology and LBC, conventional cytology and cell block, and LBC and cell block was 0.769, 0.791, and 0.945, respectively. The most prevalent malignancy which led to peritoneal fluid involvement was papillary serous carcinoma of the ovary. Conclusion: Compared to conventional cytology, the liquid-based method had no significant superiority in the evaluation of the peritoneal fluid method, but the combined usage of smears and cell block improved the diagnostic accuracy of the peritoneal washing samples in different gynecological malignancies.

Liquid-based cytology versus conventional cytology in fine-needle aspiration cytology of thyroid, salivary gland, and lymph node swellings: a comparative study

Purpose To compare the cytomorphologic features of liquid-based cytology (LBC) and conventional preparation (CP) smears in fine-needle aspiration of thyroid, lymph node, and salivary glands. Patients and methods The study included 110 patients experiencing thyroid, salivary gland, and lymph node swellings, who presented to the Surgical Oncology Outpatient Clinic and referred to Cytology Unit of the Pathology Department of the National Cancer Institute, Cairo University, for fine-needle aspiration cytology, from the period January 2017 till July 2018. For each fine-needle aspiration cytology, three passes were performed: the first two were for conventional cytology, and the third one for LBC, where a semiautomated SurePath technique was used. The comparison between both techniques was done using Wilcoxon signed rank test, regarding the cellularity, cell architecture, monolayering, nuclear details, cytoplasmic details, informative background, and bloody background. Results CP was significantly better regarding the cellularity, cellular architecture, and informative background of thyroid lesions, whereas LBC was better when the absence of bloody background was considered. On the contrary, no significant difference was found between both preparations regarding monolayer formation and nuclear and cytoplasmic details. Conclusion CP is superior to LBC in preservation of cellular pattern, gives better cellular yield, eliminates hemorrhagic background, and maintains the stroma and background, which are important for diagnosis. As such stroma is lacking in LBC, it should be interpreted with support of CP in certain lesions.

Liquid Paraffin as a Rehydrant for Air Dried Buccal Smear

Aim: To assess the efficacy of liquid paraffin as a rehydrant for air dried buccal smear based on the nuclear and cytoplasmic details. Background: Buccal smear is useful for diagnosing Malignancy, Fungal infection, Viral infection and Vesicullobullous dermatoses. The routine practice is to wetfixthe smear and send them to the laboratory for staining and evaluation by a cytopathologist. But drying of smears is inevitable, especially when the aspirate is less and when there is improper fixation. This can cause drying artifacts. An alternative method for overcoming this problem is intentional air drying followed by rehydration. Many rehydrants have been experimented upon. The most common rehydrant being saline. The present study is done to check the efficacy of liquid paraffin as a rehydrant. Material and methods: 2 sets of 20 buccal smears were collected from the patients. One set air dried for 24 hours and the other wet fixed. Conventional pap staining was then carried out. The slides were examined by two observers for preservation of Nuclear and cytoplasmic details based on the semi quantitative scoring system. Results: Excellent nuclear details were seen in 45% of air dried smear compared to 25%in routine wet fixation. The cytoplasmic details of air dried smear and routine wet fixed smear was the same. Conclusion: Liquid paraffin as a rehydrant has shown promising results and the nuclear details were found to marginally better than in routine wet fixed smear. Air dried buccal smears can be used in routine practice especially in rural areas where there is limited access to laboratories and for mass screening.

Comparative Analysis of Liquid based and Conventional Cytology Smears in Fine Needle Aspirates from Breast Lesions.

Context:Liquid-based cytology (LBC) is being extensively used for the evaluation of both gynecological and non-gynecological specimens. Suspension of cells in monolayer makes better morphological assessment possible. Along with this, inherent morphological changes such as altered, reduced, or lost background material, fragmented cell clusters, smaller cell size, nucleolar prominence, etc., need to be considered.Aim:Present study was aimed at comparative evaluation of utility of LBC versus conventional smear (CS) in assessing breast lesions and whether it can be used as an alternative to conventional preparation.Settings and Design:Present study was a prospective study in which 75 cases of breast fine-needle aspiration cytology from patients with palpable breast lumps constituted the study group.Material and Methods:The first pass was used for CS and LBC; a second pass was given. The representative CS and LBC smears were compared using several criteria.Statistics:Each feature was scored individually and evaluated statistically using Wilcoxon's signed rank test on the SPSS program.Results:A statistically significant difference was found in informative background and background blood-debris, whereas the difference was not statistically significant in other features such as cellularity, cytoarchitectural pattern, presence of monolayer, and nuclear and cytoplasmic details.Conclusion:LBC is a promising technique in the field of cytology. It has the potential to decrease the number of slides screened per case and decrease the turn-around-time.

Cocktail of Periodic Acid-Schiff and Papanicolaou: Novel staining technique for the identification of leukemic eosinophils - A pilot study.

Background:Tissue eosinophilia may be caused due to reactive, neoplastic or idiopathic reasons. Reactive eosinophils in allergic and inflammatory conditions are transient and recruited from the circulation in response to various stimuli, whereas neoplastic eosinophils of leukemias and hematological malignancies are involved in the pathogenesis of the disease. The differentiation of reactive from neoplastic eosinophils has a serious implication on the treatment and prognosis of diseases. However, both these types of eosinophils display variation in morphology and staining characteristics in routine histopathology leading to a diagnostic dilemma.Aim:The aim of this study is to evaluate the efficacy of special stains for the demonstration of eosinophils in normal/reactive lesions and leukemias.Methodology:A retrospective study comprising twenty histologically diagnosed cases each of reactive oral lesions and leukemias were obtained from institutional archives. These tissue sections were subjected to staining with routine and special stains – Carbol chromotrope, Congo red, Leishman's stain, Periodic Acid–Schiff-papanicolaou (PAS-PAP) and PAS. Statistical analysis was performed using Pearson's Chi-square test to compare the various parameters in the evaluation of the staining efficacy.Results:Carbol chromotrope and Congo red staining showed increased staining efficacy in normal/reactive eosinophils while PAS-PAP followed by PAS and Leishman's stain showed enhanced features such as homogenicity, specificity, increased staining intensity, enhanced nuclear and cytoplasmic details in leukemic eosinophils.Conclusion:Combined PAS-PAP is a novel and cost-effective staining technique in differentiating reactive and leukemic eosinophils. It is significant in recognizing leukemic eosinophils of routine biopsies and alerts the clinician to rule out any underlying malignancies.

Liquid dish wash solution – can it be an alternative in future for the expensive and hazardous xylene in hematoxylin and eosin staining of paraffin sections

Introduction: Xylene is the most commonly used clearing agent as it has excellent deparaffinising and clearing properties. Commercial xylene is a clear, colourless liquid that has a sweet and aromatic odour. However, xylene is not only expensive but also has detrimental effect on human health such as hepatitis, chemical pneumonitis, depression, anaemia, etc. Hence in this study we are replacing xylene with commercially available liquid dish wash solution (LDW) which is not only cheap but also non biohazardous. Aims: To assess the efficacy of LDW with xylene as a deparaffinising and clearing agent in routine hematoxylin and eosin staining (H&E). Materials and Methods: Blocks from 20 consecutively submitted samples at RL Jalappa Hospital between Jan – June 2015 each of breast, Cervix, Lymphnode, Fat, Salivary Gland, Muscle and Alimentary Tract was taken and two sections of 4-micron thickness were cut and deparaffinised using Liquid Dish Wash and Xylene respectively followed by H & E Staining. The 280 sections of the 140 blocks were evaluated and scored by two pathologists independently for nuclear staining, cytoplasmic staining, uniformity, clarity and crispness. Results: In the samples evaluated LDW seems to be on par with xylene cleared sections in terms of nuclear and cytoplasmic detail, clarity and crispness. However Xylene cleared sections showed slightly better uniform staining. However both sections were equally good for reporting. Conclusion: Liquid dish wash solution is an excellent alternative to xylene in H&E Staining. However more extensive studies need to be done on other types of tissues for validation

Natural substitutes for formalin: A boon to histopathology!!

Introduction:In routine tissue processing, formalin has been proved as efficient as fixative since inception and hazards associated with it are of major safety and health concern.Aim:The aim of this study is to compare the efficacy of natural fixatives such as jaggery and Khandsari over formalin.Materials and Methods:Ninety normal tissue specimen collected during minor oral surgical procedures were included in this study. Thirty specimen each were fixed in 30% jaggery (Group-A), 30% Khandsari (Group-B) and 10% formalin (Group-C). The slides obtained after tissue processing were analyzed for the quality of fixation. The tissue sections were assessed for cellular outline, cytoplasmic details, nuclear details, staining quality and overall morphology. Each criterion was rated on a scale of 1–4. (one for poor and four for excellent). The study was double-blinded and subjected to the Kruskal–Wallis ANOVA and Chi-square test.Results:The cellular outline is excellent in 90% (Group-C) followed by 36.67% (Group-B) of specimens. With respect to cytoplasmic staining 83.33% (Group-C) of tissues showed excellent results followed by 60% (Group-B), 33.33% (Group-A). Nuclear details are excellent in 86.67% (Group-C) followed by 83.33% (Group-B), 36.67% (Group-A) of specimens. With respect to staining quality 93.33% (Group-C) followed by 50% (Group-B), 26.67% (Group-A). Overall morphology is excellent in 90% (Group-C) followed by 46.67% (Group-B).Conclusion:In the present study, Khandsari was on par with formalin and our effort of using Khandsari and jaggery for tissue fixation in human oral tissues yielded good results.

Cytomorphological Differences between Liquid-Based Cytology and Conventional Smears in Fine-Needle Aspirates of Thyroid Lesions.

Background:Liquid-based cytology (LBC) is becoming an independent processing modality for cytology specimens. Decreased obscuration, single-slide examination, lesser screening time, and potential application of ancillary techniques are the various advantages it offers.Aim:To study and compare the cytological features of fine-needle aspirates from thyroid swellings on LBC with conventional smears (CS).Materials and Methods:Fine-needle aspiration was performed on 150 patients with thyroid nodule. The aspirate was first used to prepare a minimum of two CS and the remaining aspirate was used to prepare one SurePath smear. The final diagnosis was given according to the Bethesda system for reporting thyroid cytopathology (2007). Cytomorphological parameters were semi-quantitatively scored. Pearson's Chi-square test was used and P value was calculated. A P value <0.05 was considered significant. Cytohistological correlation was done wherever possible.Results:LBC showed higher nondiagnostic rate than CS. Significant cytomorphological differences on LBC included: (i) higher frequency of single, naked nuclei; (ii) lesser nuclear and cytoplasmic details; (iii) decreased colloid which appeared differently (as napkin fold and dense droplet); (iv) increased cyst macrophages; and (v) decreased obscuration by blood.Conclusion:LBC can supplement CS but cannot replace it.

Xylene/Chloroform Free Microwave Tissue Processing and Staining a Non-hazardous and Time Effective Alternative

Introduction: Tissue processing using xylene/chloroform has been employed in histopathology reporting for the past 100 years. Microwave technique has not only reduced the processing time from 1 day to one hour but also doesn’t use xylene/chloroform and has been found to be on par with conventional processing. Xylene is expensive and detrimental to human health. The present study replaces chloroform with isopropanolol in tissue processing and xylene with liquid dish wash solution (LDW) in staining which is not only cheap but also non bio hazardous. Aim: To assess the efficacy of xylene free processing (microwave) and staining versus conventional tissueprocessing and hematoxylin and eosin are staining. Materials and methods: Two Tissue bits from 15 consecutively submitted samples at RL Jalappa Hospital after 1st Jan 2015 each of breast, cervix, lymph node, fat, thyroid, skin, alimentary tract, muscle, salivary gland, liver and kidney were taken and one was processed and stained using conventional method, other using xylene/chloroform free processing and conventional staining and other with both xylene/chloroform free processing and staining. The 495 sections were evaluated and scored by two pathologists independently for nuclear staining, cytoplasmic staining, uniformity, clarity and crispness. Results: In the samples evaluated xylene free processing and staining seems to be on par with conventional xylene processed and stained sections terms of nuclear and cytoplasmic detail, clarity and crispness. However Xylene cleared sections showed slightly better uniform staining (p value->0.05) Conclusion: Xylene/chloroform free processing and staining is not only a rapid but also safe alternative to conventional processing and staining using xylene. However more extensive studies need to be done on other types of tissues for validation

Spritzer: For diagnostic cytopathology.

Exfoliative cytology is an easy, economical, noninvasive, and feasible method for early detection and screening program of any premalignant or malignant lesion. In case of routine cytological procedure, classical Papanicolaou (PAP) stain is widely used while Romanowsky stains are sparingly used. Leishman-Giemsa (LG) cocktail, being a easier, cost effective staining technique, has not been used in exfoliative cytology. Therefore, this pilot study was carried out to compare and contrast the role of LG stain in routine cytological procedure which is very cost-effective, less time-consuming and requires less infrastructural support. To study the diagnostic efficiency of LG cocktail in comparison with PAP stain and Feulgen stain in mucosal cells for evaluating cellular changes of oral squamous cell carcinoma (OSCC). Three cytological smears were prepared from 10 healthy controls and 10 patients clinically diagnosed with OSCC, and they were stained with LG cocktail stain, PAP stain, and Feulgen stain. The stained smears were evaluated for cytologic diagnosis and the staining characteristics such as nuclear and cytoplasmic details were recorded as per criteria by Sujathan et al. The data were statistically evaluated with analysis of variance test using SPSS 15 software for windows. The results from the cases diagnosed as OSCC by PAP and LG cocktail were almost identical and superior to Feulgen stain both in diagnostic ability and in staining characteristics. The one-step LG cocktail is easy, very cost-effective, less time-consuming with less infrastructural support as compared to PAP stain; however, it warrants further evaluation for screening of oral cancer as a potential aid.

Comparitive efficacies of a natural fixative with a conventional fixative.

Introduction:The quest for formalin substitutes has long been going on due to its health hazards. Honey has been recognized as a safe substitute for formalin. However, we explored jaggery as a natural substitute for formalin. The aim of this study was to compare the tissue fixation abilities of jaggery syrup (30%) with that of 10% neutral-buffered formalin (NBF) and to determine the best fixative among both.Materials and Methods:A study was conducted with 65 pathological tissues. Each specimen was divided into two equal parts. One part was fixed in 30% jaggery solution (Group A), while the other half was fixed in 10% NBF solution (Group B). 24 h tissue fixation was attained at room temperature followed by evaluation of pre- and post-fixation, tissue shrinkage, weight difference and ease of sectioning, followed by evaluation of conventional processing and staining. The histomorphological assessment for each slide was made based on evaluation of cellular outline, cytoplasmic details, nuclear details, staining quality and overall morphology under light microscopy. Each criterion was rated on a scale of 1–4. Nominal categorical data between the groups were compared using Chi-squared test.Results:The preservation of tissue specimen by jaggery syrup was comparable to that of formalin and surprisingly overall nuclear detail of the tissue was better than conventional formalin fixative.Conclusion:Jaggery can be successfully adopted in routine histopathology laboratories in place of formalin.

Leishman-Giemsa Cocktail - Is it an Effective Stain for Air Dried Cytology Smears.

Air dried cytology smears are stained routinely with Romanowsky stains so that the relative cell size, nuclear size, cytoplasmic details, smear background elements and intercellular matrix components are better appreciated. A variety of modified Romanowsky stains are used in cytology. Leishman-Giemsa (LG) cocktail is one of the new staining techniques which can be used for staining the air dried cytology smears. To evaluate the quality of staining of LG cocktail on air dried smears and to compare the quality of staining of LG cocktail with May Grunwald Giemsa (MGG) which is the most commonly used stain in cytology. The present prospective comparative study was carried out with 100 cases and two extra smears were prepared for each case and stained with MGG and LG cocktail stains. The stained slides were blinded and were evaluated for the staining characteristics of the nucleus, cytoplasm and background staining. Based on this, scoring was done by two pathologists independently. Quality Index (QI) was calculated by dividing the scores obtained with the total score possible. LG cocktail stained slides were excellent in cytoplasmic staining, granularity, nuclear morphology, background material staining and overall staining characteristics. QI of LG cocktail was 0.8 while that of MGG was 0.59. Staining of air dried smears by LG cocktail has a good QI. It is also cheaper, requires short duration for staining as compared to MGG. Hence, LG cocktail can be an effective replacement for MGG for staining the air dried cytology smears.

Diagnostic Utility of Cell Block Method versus Cytospin Method in Pleural and Peritoneal Fluid Cytology

Background: Cytological evaluation of body cavity fluid is diagnostically challenging. Improved ethanol formalin fixative is used which offer excellent cytomorphological features. Cell blocks prepared from residual tissue fluids or effusion obtained by aspiration, can be useful adjunct to smear for establishing a more definitive cytopathologic diagnosis. . Methods: A total of 170 fluid specimens were examined for cytospin smear and cell block method. Out of 170 fluids, 102 were peritoneal and 68 were pleural. Each fluid specimen was subjected to cytospin smear (CSS) technique, and 10% alcohol-formalin cell block (CB) technique. Overall morphological details, cellularity, architecture, nuclear and cytoplasmic details were studied in both CSS and CB techniques. Results: In this study, analysis body fluid specimens using cytospin smear and cell block methods revealed that there is no difference between cytospin smear method and CB in defining the benign, fungal and inflammatory conditions. However, CB method could able to identity papillary pattern more efficiently than the cytospin method. Conclusion: Although there was no statistical difference between the results obtained by the cytospin and cell block methods, cell block method in our study accurately diagnosed the cases which were missed or incompletely diagnosed on cytospin smear method. Thus cell block proved to be superior method for the study of effusion as compared to cytospin smear. As the cell blocks permit longer storage and additional analysis such as immunohistochemistry (IHC) and microarray, it should be adopted additionally for