COMPARISON OF DIFFERENT STAINING METHODS FOR ANALYSING ESTERASE (EST) ISOZYME OF FUNGI AND PLANTS

EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong different stain recipes for Est of 3 kinds of fungi-Lentinus edodes. Pleurotus sapidus. Phellinus igriarius and 2 kinds of plants-Populus sp and Brassica chinensis. Of the four kinds of Est staining recipes tested.the recipe α-acetic acid-naphther showed the best effect.and followed by β-aceticacid-naphther, semicontent α-aceticacid-naphther and α+β-aceticacidnathpher.

A RAPID STAINING METHOD SHOWING AgNOR AND DNA IN PATHOLOGICAL DIAGNOSIS

In order to satisfy the need of rapid clinical diagnosis, a series of experimental studies for AgNOR (Argyrophilic NORassociated proteins) and DNA (Dereyribonucleic acid) staining was compared, from which a stable staining solution was selected to show mucin residence. Using the referred tables, with reference to the corresponding relation of the staining solution with temperature and time, reliable staining results for AgNOR and DNA could be obtained.These two modified staining methods are simple, reliable and easy to operate, able to provide good contrast for pathological diagnosis.

A Comparative Study on Four Staining Methods for Antitumor Active Fraction of Periplaneta americana

[ Objective ] This study aimed to compare four staining methods for proteins of SDS-polyacrylamide gel dectrophoresis （SDS-PAGE） and explore a suit- able staining method for the antitumor active fraction of P. americana after polyacrylamide gel electrophoresis. [ Method ] BSA was used as the standard for the comparison of Coomassie brilliant blue staining method, potassium staining method, calcium staining method and silver staining method, on the basis, antitumor ac- tive fraction samples of P. americana were used for SDS-PAGE electrophoresis and staining. [ Result] The results showed that silver staining method could be ac- curately, quickly and easily used for SDS-PAGE staining of the antitumor active fraction of P. amer/cana. [ Conclusion] This study laid the foundation for explo- ring the medicinal value of P. americana.

Introducing a Rapid and Safe Method for Myeloperoxidase Staining

Background: Myeloperoxidase staining is used to differentiate leukemias since several decades. Despite implementation of flow cytometric, cytogenetic and molecular techniques for identification of leukemic blasts, histochemical stains such as myeloperoxidase stain are persistently used for better classification of leukemias. The myeloperoxidase staining is a time consuming and hazardous procedure. The present report describes a sensitive, rapid and easy method for assessment of peroxidase activity. Materials and Methods: Bone marrow aspiration slides were stained with Dako product: Code number: K3467 containing DAB chromogen (3,3-diaminobenzidine in chromogen solution) and substrate buffer (Imidasole-HCL buffer, PH 7.5 containing hydrogen peroxide and an anti microbial agent) in a rapid procedure taking only ten minutes time. The staining needs no material preparation steps. Neutrophils in the slide are taken as positive control or another normal smear was costained to be used as control. All cases were followed up with flow cytometry and cytogenetic studies. Result: The reaction product of this stain is brown and granular. Promyelocytes and myelocytes are the most strongly staining cells with positive (primary) granules. Lymphoblasts are negative. The result of classification of leukemias with this technique was in concordance with flow cytometric immunophenotyping. Discussion: Many practical techniques have been described using benzidine as an indicator for myeloperoxidase staining. Benzidine is a carcinogenic material and its usage is severely restricted in laboratory. Formerly we prepared requisite materials for myeloperoxidase staining by hazardous ways (boiling), but we decided to apply ready to use 3,3-diaminobenzidine (DAB), which is used in final step of immunohistochemistry stains. Conclusion: Use of 3,3-diaminobenzidine (DAB) is highly recommended for myeloperoxidase staining, while the result is extraordinary and fully compatible with flow cytometry and the method is safe and rapid.

A NEW STAINING METHOD FOR THE MEASUREMENT OF PROTEIN USING TETRAPHENYLPORPHYRIN TETRASULFONATE(TPPS_4)

A new dye-staining method for protein assay is described.The reaction between TPPS_4 and protein molecule causes a shift in the absorption of TPPS_4 from 435 nm to 488nm.The absorbance at 488 nm is proportional to the concentration of protein.The range of Beer's law was 1-5 ug/ml and the Sandell's sensitivity was 0.0087 ug/cm^2.

利用Hoechst 33342-PI荧光双染法评价氰氨化钙对茄病镰刀菌生物活性的影响

本试验以茄病镰刀菌(Fusarium solani)为研究对象,探索了Hoechst 33342及PI荧光染剂对病菌不同活性孢子的色泽差异,明确了病原菌经Hoechst 33342染色后的活孢子发蓝色荧光,最佳染色浓度和时间为1μg/mL、20 min;碘化丙啶(PI)染色后的死孢子发红色荧光,最佳染色浓度和时间为3μg/mL、10 min。并由此建立了Hoechst 33342-PI荧光双染技术,应用该技术可快速、清晰地甄别出不同浓度氰氨化钙对茄病镰刀菌孢子活性和萌发的影响,结果表明,氰氨化钙对茄病镰刀菌的灭杀效果随着浓度增高而增强,且与处理时间呈正相关。2000 mg/L氰氨化钙处理3 h,孢子致死率高达83.97%;500~1000 mg/L处理3 h,孢子致死率为7.15%~26.80%,孢子萌发率为2.40%~3.59%,处理48 h时,孢子致死率达92.07%~100%。250 mg/L氰氨化钙对茄病镰刀菌的致死效果微弱或没有影响。本研究通过菌丝速率法和生物量法进一步验证了应用Hoechst 33342-PI荧光双染法评价氰氨化钙对茄病镰刀菌孢子活力的影响是可行的。这为突破植物病原微生物药剂室内常规筛选方法提供了新的思路和技术手段。

包头地区1 249例浅部真菌病患者中荧光染色法和KOH湿片法应用对比研究

目的:分析比较荧光染色法和KOH湿片法检测浅部真菌病的效果及不同位置检测结果比较。方法:收集2021年10月至2023年3月在包头医学院第二附属医院皮肤科就诊拟诊断为浅部真菌病患者的标本,分别采用荧光染色法和KOH湿片法直接镜检,比较阳性率检出结果,并进一步针对不同检测部位检测阳性率比较分析。结果:KOH湿片法和荧光染色法分别检测558例和691例患者(阳性率分别为38.35%和58.47%),阳性率差异有统计学意义(P<0.001);且头面部、手部、足部、躯干、腹股沟及甲部位荧光染色法阳性检出率均高于KOH湿片法,差异有统计学意义(P<0.05),差异由大到小依次为足部、头面部、腹股沟、手部、甲部、躯干部(P<0.001)。结论:荧光染色法在浅部真菌病的实验室检验中阳性检出率明显优于KOH湿片法,可以降低临床漏诊误诊率,在甲部、足部、腹股沟、头面部的检测中可作为优选,手部、躯干部则根据临床医生的临床经验在KOH湿片法或荧光染色法中选择,以减轻患者经济负担。

Two New Identification Methods for Encephalitozoon cuniculi on Tissue Section

[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathological sections of sick rabbits were stained and identified. [ Result] The pathological changes in brain tissue could be clearly observed on sections, but parasites were not examined in pathological brain tissues stained by common staining method. When the pathological section was stained by improved Gram staining method, the pathological changes in brain tissue were not ouly stained very clearly, but blue parasites were also found in brain tissues. The parasites in epithelioid cells were stained into purple ones by methyl green pyronin staining method. [ Conclusion] The im- proved Gram staining method and methyl green pyronin staining method performed good staining effects of E. cuniculi in pathological sections, which were conducive to rapid diagnosis of encephalitozoonosis in rabbit.

Comparative Study of Z N Staining vs. Flurochrome Staining and Impact of Sample Processing on Diagnosis of Tuberculosis from Various Clinical Samples

Background: Tuberculosis is a highly infectious disease and India has the highest burden with it. Diagnosis of tuberculosis in many countries is still dependent on microscopy. Although its sensitivity is low in comparison to culture and molecular methods, its sensitivity can still be improved by using fluorescence staining method and processing of samples by homogenization and concentration method. Material and methods: Samples were collected from all newly registered suspected cases of tuberculosis in tertiary care hospital from outward and indoor department during a period of one year. Smears were prepared for Ziehl Neelsen stain and fluorescence stain both before and after homogenization and concentration procedure by 4% NAOH-2.9% sodium citrate method and results of them were interpreted according to RNTCP criteria for grading of sputum samples. All the samples were cultured in liquid culture MGIT system (Mycobacterial Growth Indicator Tube) and results of microscopy were compared with liquid culture taken as gold standard. Data were analyzed by using SPSS software version 16. Result: 350 samples were collected during study period. Out of 350 samples, 48 samples were positive for M. tuberculosis by MGIT system. In comparison with MGIT system, sensitivity of Z N stain for detection of acid fast bacilli was 77% before decontamination procedure, which was increased up to 85.42% after decontamination and concentration process. Sensitivity of fluroscence stain was 85.42% before processing, which was increased up to 91.67% after processing of samples. Conclusion: Sensitivity of smear microscopy can be enhanced by use of fluroscence microscopy and concentration method.

免疫荧光检测平台联合快速HE染色在阴道炎中的诊断价值

目的分析免疫荧光检测平台联合快速HE染色在阴道炎中的诊断价值。方法选取2021年1月至2022年12月在首都医科大学附属北京妇产医院怀柔妇幼保健院妇科就诊的250例疑似阴道炎患者为研究对象。以实验室检验作为金标准,分析不同检验方式检测阴道炎的诊断效能及阴道清洁度检出情况。结果免疫荧光联合快速HE染色法的诊断效能高于湿片镜检法,差异有统计学意义(P<0.05);免疫荧光联合快速HE染色与湿片镜检法的阴道清洁度检出情况比较,差异无统计学意义(P>0.05)。结论免疫荧光检测平台联合快速HE染色在阴道炎中的诊断价值较高,能够提高诊断效能。

荧光染色法与KOH湿片法在皮肤真菌直接镜检检查中的应用价值

目的对比荧光染色法与氢氧化钾(KOH)湿片法用于皮肤真菌直接镜检检查中的效果。方法以2022年10月至2023年10月我院接诊疑似皮肤真菌感染患者90例作为研究对象,采集皮损样本,分别通过荧光染色法、KOH湿片法进行镜检检查,以临床综合诊断结果为金标准,对比两种检查方法的阳性检出率。结果临床综合诊断结果显示,31例患者确诊为手足癣,占比34.44%;19例患者确诊为体股癣,占比21.11%;11例患者确诊为头面癣,占比12.22%;20例患者确诊为甲癣,占比22.22%。荧光染色法直接镜检取得的总阳性率高于KOH湿片法(P<0.05)。结论通过荧光染色法进行皮肤真菌直接镜检检查相比KOH湿片法结果更准确,且操作更为便捷、快速。

Test Method for Evaluating the Cleaning Performance of Proteases in Automatic Dishwashing Detergent

A test method was developed to better evaluate the cleaning performance of protease in Chinese automatic dishwashing detergent(ADD)and differentiate the washing performance of different ADD formulations on protein stains.Steamed egg stain,which was a Chinese-consumers-relevant soil,was used as monitor.The response curves of the stain to different amounts of protease were measured under laboratory conditions,and the stain removal ability of commercially available ADDs was tested with this method.The results showed that the soil removal rate of ADD without protease was less than 10%,and the soil removal rate increased with the increase of protease dosage in the range of 0~2.0%.Steamed egg stain had a good response to protease,and it could effectively distinguish the cleaning performance of commercial ADDs.The test method could be used in the development,screening and evaluation of ADD formulations.

HE染色光镜观察法在脑组织形态学研究中的应用

HE染色光镜观察法是生物、医学基础研究中最基本的研究方法之一。通过复习整理针刺干预对相关神经系统疾病大鼠模型病理变化的影响,并与免疫组化染色法比较,了解HE染色方法在中医针灸作用机理研究中的地位和作用。表明HE染色方法可以在同一个视野下同时观察大鼠脑组织的多种指标变化情况,以整体判断大鼠脑的功能状态,和免疫组化染色法相比具有观察直观,操作简便易于掌握,而且经济节省时间的特点,明确HE染色光镜观察法作为一种最基础的方法在脑组织形态学的研究中有着其不可替代的优势。

芒(Miscanthus sinensis Anderss)花粉生活力测定方法比较

采用离体萌发法、FDA染色法和I2-KI染色法测定芒离体花粉的生活力,并对测定效果进行比较。结果表明,利用离体萌发法测得芒离体花粉的平均初始萌发率为82.6%,并且花粉萌发和生长很快,5min即开始萌发,培养30min后花粉管平均长度达到了145.77μm,但花粉的平均萌发率下降很快,室温保存90min的花粉其萌发率已下降至3.0%。利用FDA染色法和I2-KI染色法测定的芒花粉初始生活力与离体萌发法的结果基本一致,分别为84.6%和86.6%,但这两种染色法在跟踪测定芒离体花粉的生活力时易误判,不适合用于跟踪测定花粉生活力的变化。利用离体萌发法能够准确有效地测定芒花粉生活力的变化规律,而FDA染色法和I2-KI染色法适合用于测定芒花粉的初始生活力。

HE染色细胞核灰染的处理方法比较研究

目的探讨HE染色细胞核灰染的处理方法,提高HE染色质量。方法收集10例HE染色发灰的胃粘膜、肠息肉和子宫内膜等不同类型的组织,重新切片,通过调整苏木素染色条件以及染色前对组织进行修复处理,设对照组,比较各种处理方法的染色效果。结果通过延长时间、水浴加热和微波加热等调整苏木素染色条件的方法以及水煮修复处理法能够不同程度地促进核染色,但灰染现象未能彻底解决。PBS、EDTA、柠檬酸分别采用高压和微波修复处理均能较好地改善HE染色细胞核灰染现象,以PBS微波处理法效果最佳。结论 PBS微波处理法是HE染色细胞核灰染现象的一种有效处理方法。

骨髓活检HE染色时细胞核灰染的处理方法比较

目的探讨骨髓穿刺活检组织在HE染色时细胞核灰染的处理方法,以提高染色质量。方法收集HE染色时骨髓穿刺活检组织中细胞核灰染的病例15例,每例重新切片,通过延长苏木素染色时间、高压修复、微波修复及氨水浸泡等4种不同的处理方法,设对照组,比较各种处理方法的染色效果。结果高压法、微波法和氨水法均能明显改善骨髓腔内各细胞核的灰染现象,但高压法易使骨小梁脱片,微波法处理时间较长,氨水法既能保持组织结构的完整性,又用时较短。延长时间法改善细胞核灰染的效果不及前三种方法。结论氨水法是处理骨髓穿刺活检组织在HE染色时细胞核灰染的一种有效的方法,且操作简单、快速。

Ziehl-Neelsen-维多利亚蓝-碘绿复合染色法显示抗酸杆菌

目的 :探讨显示皮肤活检组织中抗酸麻风杆菌、弹力纤维和细胞等组织成分的复合染色法。方法 :运用Ziehl Neelsen(Z N)苯酚复红染色液、维多利亚蓝 (Victoriablue ,VB)染色液、碘绿 (iodinegreen ,IG)和马汀氏黄 (Mar tiusyellow ,MY)染色液对皮肤活检组织进行复合染色。结果 :组织内结核样结节中麻风杆菌显示为红色 ,细胞核呈淡绿色 ,细胞浆无色 ,结节周围的弹性纤维呈蓝色 ,基质呈淡黄色。结论 :Z N原法染色单一 ,对比效果差 ,应用碘绿、马汀氏黄和维多利亚蓝复合的改造染色液 ,能更好显示组织中的麻风杆菌 ,以及包绕于结核样结节周围的弹力纤维等成分 ,该方法是一种对比清晰 。

PDCA循环方法在HE切片质量持续改进中的应用

目的观察HE切片常见问题及质量控制效果。方法收集2018年1月-10月某院科室切片500张作为研究对象,对基本资料进行分析并且根据分析结果应用PDCA循环法持续改进HE切片质量,观察实施PDCA循环法后的切片质量。结果切片不完整67张(13.40%),切片过厚70张(14.00%),切片刀痕46张(9.20%),切片起皱与压缩60张(12.00%),切片离刀与粘在蜡块上80张(16.00%),切片破碎50张(10.00%),切片入水散碎65张(13.00%),切片时组织污染30张(6.00%),切片染色不均匀32张(6.40%);实施PDCA循环法后切片质量评分高于实施前,差异有统计学意义(P<0.05)。结论 HE切片存在质量问题较多,其中以手工操作问题为主,实施PDCA循环法措施可有效降低HE切片不良质量问题发生率,值得临床进一步推广应用。

不同冻存法对肌肉冰冻切片后HE染色和肌球蛋白ATP酶染色效果的影响

使用分步法和一步法冻存巴马香猪背最长肌,冰冻切片后比较苏木精-伊红染色法(HE染色)和肌球蛋白三磷酸腺苷酶染色法(ATP酶染色)效果。结果表明:分步法冻存的肌肉样品冰晶少,细胞形态完整,HE染色效果好,核质明显;ATP酶染色效果好,可较好地区分Ⅰ型Ⅱ型肌纤维。一步法冻存的肌肉样品冰晶多,细胞形态不完整,HE染色效果差,核质模糊;ATP酶染色效果差,基本染不上色,无法对肌纤维类型进行区分。分步冻存法的建立为通量式制备猪肉的恒冻切片奠定了基础,也为进一步开展相关性状的生理学、遗传学及种猪选育提供了很好的表型测定方法。

应用荧光染色剂Fluorescent Brightener 28和Propidium Iodide染色识别家蚕微孢子虫

家蚕微粒子病被列为蚕种生产的唯一法定检疫对象,母蛾镜检家蚕微孢子虫(Nosema bombycis,Nb)是主要的检疫方法,但其准确性受到多种因素影响。为探究应用荧光增白剂28(Fluorescent Brightener 28,FB28)和碘化丙啶(Propidium Iodide,PI)染色识别Nb孢子的可行性,以感染Nb的家蚕幼虫中肠、蚕卵、母蛾、蚁蚕为材料,联合应用以上2种荧光染色剂进行Nb孢子检测效果试验。在荧光显微镜下可见成熟的Nb孢子被FB28染成蓝色且细胞核被PI染成红色,而未成熟的孢子只能被PI染色。采用2种荧光染色剂染色,在被感染的幼虫中肠、蚕卵、母蛾、蚁蚕中都能够检测到Nb孢子,并且带毒母蛾研磨液样品稀释1 000倍依然可以检测到Nb孢子。幼虫中肠感染后1~2 d很难观察到Nb孢子,感染后3 d可以观察到局部存在少量未成熟的Nb,感染后4 d开始局部出现成熟的Nb,至感染后5~6 d可见组织中大面积分布有成熟孢子和未成熟孢子。荧光显微镜下观察被染色的Nb孢子的细胞核明显小于幼虫中肠细胞核,且亮度明显较强,卵壳、母蛾、蚁蚕表面的几丁质碎片形状与Nb孢子明显不同,并且不能被PI染色。试验结果表明,该染色检测方法可以排除蚕体组织细胞、几丁质碎片的干扰,提高对Nb检测的准确度,并且在Nb浓度较低的情况下,可以提高检测的灵敏度。