LEISHMAN GIEMSA COCKTAIL, A CYTOLOGICAL STAINING COMPARABLE TO PAPANICOLAOU STAIN FOR ORAL CANCER DIAGNOSIS.
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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.
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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.
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Milk smears from samples of mastitis affected milk were stained by both Papanicolaou stain and Leishman stains to study the comparative efficacy of these stains in cytological study of bovine mastitis. It was observed that with Papanicolaou stain, the constituent cells showed remarkable clarity, transparency and differential staining as compared to Leishman stain. Cytoplasmic acidophilia and pyknosis were characteristically seen in degenerated cells with Papanicolaou stain. It will help in correct assessment of nature of infiltrating cells and effectiveness of host response. Thus, Papanicolaou stain could be the stain of choice for cytological study of bovine mastitis.
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Leishman stain, a type of Romanowsky stain, is a mixture of Methylene blue and Eosin dye.It is prepared in alcohol and is diluted using distilled water.It stains human cells purple in colour.It is one of the best methods preferred for peripheral blood smear examination.Giemsa stain has been named after a German chemist and bacteriologist Gustav Giemsa and used in histopathological diagnosis of malaria and various other parasites.Comparative study in between Leishman's stain and Giemsa stain was done.Randomly selected 20 peripheral blood samples were used for this study.Two Peripheral smears were prepared from each sample with a total of forty smears.The first group of twenty smears was selected from each sample and stained with routine Leishman's stain and the second group of twenty smears from each sample was stained with Giemsa stain.The morphology of the blood cells was analysed by a pathologist in an unbiased manner.The scores were analysed statistically.The smears were kept for storage after mounting with DPX mounting media.This study shows that Giemsa staining is a better alternative for Leishman's staining for a routine peripheral smear examination and this also has advantages of finding other abnormalities in blood such as other blood element abnormalities.Due to no significant difference seen in staining property in between Leishman's stain and Giemsa stain, we can very well use Giemsa stain as an alternative to Leishman stain.
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Summary Introduction Peripheral blood and bone marrow smear examination is an important basic tool for the diagnosis of different haematological conditions including haematological malignancies. We created a newer modification of the conventional Leishman and Giemsa stains as Leishman and Giemsa (L&G) stain and compared the efficacy and reliability of this stain with conventional stains. The study was performed to evaluate the staining efficacy, feasibility, time and cost of L&G stain over the conventional Leishman and Giemsa stains. Methods A pilot study was carried out in the Department of Haematology of our hospital from October 2013 to December 2013. Hundred selected cases, each with peripheral blood and bone marrow smears were taken, and three sets of the smears were prepared from each sample – one for L&G stain and other two – one each for conventional Leishman and Giemsa stains. This staining is further incorporated in our routine standard operating protocols for staining of all the peripheral blood smears in automated stainer, Sysmex SP 10. Result The average grading score from each staining methods from all the three experts was compiled. The average grading score of L&G staining method was noted to be significantly higher than the other two methods (analysis of variance test, P value < 0.05). When modified L&G stain (C) was compared with stain conventional stains (A and B), a P value of <0.001 was noted in all parameters except between Leishman stain and L&G stain in mature RBC and WBC nucleus and RBC inclusions ( P value between 0.05 and 0.001). Conclusion L&G staining is a newer staining technique of immense help in high‐throughput haematology laboratories by offering a time‐saving, cost‐effective and better staining option to conventional staining methods. It gives a better nuclear and cytoplasmic differential staining and can also be used in automated blood counters/stainer.
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Background: This study was planned to compare and evaluate the staining efficacy of Leishman–Giemsa cocktail (LG), Papanicolaou, and Giemsa stain (G) in potentially malignant disorders and malignant lesions. Aims: To evaluate the quality of nuclear and cytoplasmic staining of LG with G, and rapid Papanicolaou stain (Pap) and to compare the total staining efficiency of LG against G and P. Materials and Methods: One hundred and eighty participants were studied under three groups – 60 as healthy controls, 60 with potentially malignant disorders, and 60 with malignant lesions; smears were taken thrice from the buccal mucosa. One smear was fixed with Bio-Fix spray and other two smears were allowed to air dry for 2–3 minutes. Then, the ethyl alcohol-fixed smear was stained with Pap and the two other air-dried smears were stained with G and LG stains. Analysis was done using Friedman test and Wilcoxon Signed Rank Test with SPSS Version 15.0. Results: In the normal group, staining of LG was highly significant (P < 0.001). Among potentially malignant lesions, LG was observed to be highly significant (P < 0.001) when compared with G and was not significant when compared with Pap (P = 0.186). In the malignant group, LG was highly significant (P < 0.001). LG was superior with the highest average staining score of (2.018) than Pap and G. Conclusion: LG cocktail is a better stain with excellent cytoplasmic and nuclear staining intensity compared to Pap and G stains.
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Introduction: Papanicolaou(PAP) staining is commonly used for staining cervical cytology.Leishman Giemsa (LG) cocktail, being a relatively new staining technique, is now being usedextensively in exfoliative cytology.Aims & Objective:To study and evaluate the diagnostic efficacy and reliability of LG cocktail in comparison with the PAP stain in exfoliated cells of cervical cytology.Materials &Method: Cross-sectional study conducted at department of pathology for 3 months.The pap smears were stained with LG and PAP stains.The smears were evaluated in terms of nuclear morphology,Cytoplasm and background and scored as per the criteria of Sujathanetal.Results: Sample size was 200 (100 pap stained and 100 LG cocktail stained).Both Cytoplasmic staining and Nuclear staining was better in LG. Background staining is more in LG Cocktail stain but was not obscuring the cell morphology.Conclusion: Papanicolaou staining is widely used technique but it has few limitations LG cocktail staining method is an easy, cost-effective and one-step technique that can be helpful in screening large number of cases in screening cervical cancers.
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Background: Papanicolaou staining is commonly used for staining exfoliative cytology smears with Romanowsky stains being used sparingly. Leishman Giemsa (LG) cocktail, being a relatively new staining technique, has not been used in exfoliative cytology. This easy, cost-effective and one-step technique warrants further study because of its potential application in screening of oral cancer.
Aim: To study and evaluate the diagnostic efficiency and reliability of Leishman Giemsa (LG) cocktail in comparison with Papanicolaou (Pap) and May-Grόnwald Giemsa (MGG) stains in exfoliated cells for the detection of oral squamous cell carcinoma.
Materials and Methods: Three smears were prepared from each 100 controls (buccal mucosa) and 100 patients, clinically diagnosed with oral squamous cell carcinoma and stained with Pap, MGG and LG cocktail stains. The slides were evaluated for the staining characteristics of nucleus and cytoplasm. The diagnostic efficiency of each stain was evaluated by comparing the cytologic diagnosis of each stain with the histopathological diagnosis. Finally, the diagnostic reliability was evaluated by comparing the three stains with each other and the histologic diagnosis.
Statistical Analysis: The data were statistically evaluated with Friedman test, Wilcoxon sign rank test and McNemar chi square test using SPSS15 software.
Results: The results from the histologically confirmed cases of squamous cell carcinoma and the number of cases diagnosed by Pap and LG cocktail were almost identical and both were superior to MGG. The P value obtained for the confirmed cases of squamous cell carcinoma in comparison for Pap vs MGG was 0.001, MGG vs LG cocktail was 0.001 and LG cocktail vs Pap was 0.157. Hence, no statistical significant difference was observed between the diagnostic ability of Pap and LG cocktail stains.
Conclusion: LG cocktail is an easy, cost-effective and one-step technique comparable to Pap staining; however, it warrants further study in its potential application in screening of oral cancer.
Aim: To study and evaluate the diagnostic efficiency and reliability of Leishman Giemsa (LG) cocktail in comparison with Papanicolaou (Pap) and May-Grόnwald Giemsa (MGG) stains in exfoliated cells for the detection of oral squamous cell carcinoma.
Materials and Methods: Three smears were prepared from each 100 controls (buccal mucosa) and 100 patients, clinically diagnosed with oral squamous cell carcinoma and stained with Pap, MGG and LG cocktail stains. The slides were evaluated for the staining characteristics of nucleus and cytoplasm. The diagnostic efficiency of each stain was evaluated by comparing the cytologic diagnosis of each stain with the histopathological diagnosis. Finally, the diagnostic reliability was evaluated by comparing the three stains with each other and the histologic diagnosis.
Statistical Analysis: The data were statistically evaluated with Friedman test, Wilcoxon sign rank test and McNemar chi square test using SPSS15 software.
Results: The results from the histologically confirmed cases of squamous cell carcinoma and the number of cases diagnosed by Pap and LG cocktail were almost identical and both were superior to MGG. The P value obtained for the confirmed cases of squamous cell carcinoma in comparison for Pap vs MGG was 0.001, MGG vs LG cocktail was 0.001 and LG cocktail vs Pap was 0.157. Hence, no statistical significant difference was observed between the diagnostic ability of Pap and LG cocktail stains.
Conclusion: LG cocktail is an easy, cost-effective and one-step technique comparable to Pap staining; however, it warrants further study in its potential application in screening of oral cancer.
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Background: Romanowsky stains are universally used for routine staining of peripheral blood smears.Among these Leishman stain is most commonly used in hematology laboratories worldwide.This study was done to stain peripheral blood smears with modified Leishman stain (MLS) on Day 1, Day 5, Day 10 of preparation of the stain and to assess the quality of staining by scoring the stained smears by calculating the Quality Index (QI) and comparing the scores with normal peripheral smear.Methods: Study was done in the hematology section of our institute from December 2016 to February 2017 on a sample size of hundred and one.(MLS) was prepared by adding phenol to the Leishman stain.All cases were stained with modified Leishman stain on day 1, 5 and 10 after preparing the stain.All the smears were scored based on overall staining, cytoplasmic staining, nuclear morphology, red cell staining and platelet staining.Quality index was calculated by dividing the score obtained by maximum score possible.Result: Overall staining, cytoplasmic staining, nuclear morphology, red cell staining and platelet staining were better on day 10 after preparation of MLS when compared to day 1 and day 5.The Quality Index of stained smears normal leishman stained smear was 0.95, score on day 1 of preparing stain was 0.71, score on day 5 was 0.73 and day 10 of preparing stain was 0.89. Conclusion:MLS can be used for staining of thin peripheral blood smears in 4 minutes, unlike the conventional Leishman stain method which takes about 10-15 minutes.
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