荧光染色法和传统KOH湿片法在甲真菌病诊断中的应用比较

目的探讨真菌荧光染色法与传统KOH湿片法在临床甲真菌镜检中的应用效果。方法选择2017年6月至2018年4月我院600例皮肤科门诊就诊疑似真菌感染患者指(趾)甲镜检标本,分别运用荧光染色法和KOH湿片法对这些标本进行检测,记录实验数据,并对检测结果进行比较。结果荧光染色法检测阳性为418例,阳性率为70%,其中指甲阳性130例,趾甲阳性288例。KOH湿片法检测阳性为354例,阳性率为59%,其中指甲97例,趾甲257例。指甲(χ²=8.816,P=0.003)、趾甲(χ²=7.884,P=0.005)及总阳性率(χ²=14.876,P<0.001)在两种方法中差异均有统计学意义。结论在对指(趾)甲真菌感染的诊断中,荧光染色法的阳性检出率明显高于KOH湿片法,是一种快速、准确的真菌镜检方法,值得临床推广使用。     

荧光染色法与KOH湿片法在真菌直接镜检中的应用比较

目的探索一种结果更可靠的真菌直接镜检方法。方法对600例临床高度疑似真菌感染的标本分别采用荧光染色法和KOH湿片法进行真菌直接镜检。结果荧光染色法和KOH湿片法的真菌检出率分别为97%和88.5%。结论荧光染色法是一种适合临床的、快速有效的真菌镜检方法。     

KOH试管法与玻片法在甲真菌检查中的效果比较

<正>甲真菌病是最常见的皮肤科疾病之一,发病率约占临床甲病的一半以上,病原菌包括皮肤癣菌、酵母菌和其他非皮肤癣菌性丝状真菌等,国内甲真菌病病原菌中皮肤癣菌占65%～70%,酵母菌占10%～30%。甲真菌病常表现为甲板增厚、分离、变色等临床症状,其临床表现有时不易与白甲症、黄甲综合征、甲银屑病、甲扁平苔藓等指(趾)甲疾病鉴别^[1-5],因此,病原学检查是诊断和鉴别诊断甲真菌病的关键依据。     

真菌荧光染色法与PAS染色法在肺隐球菌病病理学诊断中的比较

<正>肺隐球菌病是一种由隐球菌感染引起的肺部真菌疾病,具有传染性,近年来免疫功能正常的人群也有发病,整体发病率呈增高趋势^[1]。由于其临床症状无特异性,易与肺部其他疾病相混淆^[2]。检验科主要依靠革兰染色涂片法及真菌培养法检测,但革兰染色存在缺陷,其检出率不高,容易漏检,而真菌培养时间长,阳性率低^[3]。对于组织标本,病理科诊断该病一直以来用的都是较为传统的石蜡组织切片PAS染色法技术^[4],近年来真菌荧光染色法也作为一项比较成熟的染色方法被临床科室运用。     

KOH和乳酸酚棉蓝染色法在浅部真菌病直接镜检中的应用比较

直接镜检法是浅部真菌形态学检查的基本方法,也是最简单、快速、实用的实验室诊断方法[1].传统的方法是用10％ KOH做浮载液直接镜检,本实验用乳酸酚棉蓝染液和KOH溶液对同一患者的同一部位标本同时进行镜检,比较这两种方法的镜检阳性率. 1材料和方法 1.1标本来源 取自本院皮肤科门诊患者,拟诊为浅部真菌感染者.其中男245例,女182例,共427例.取材部位：甲部52例,头部28例,手足部163例,躯干、四肢及股部184例.     

芝加哥天蓝法在甲屑真菌检测中效果的评估

目的 探讨芝加哥天蓝染色法对甲真菌病患者标本检测效果.方法 收集皮肤科门诊确诊的甲真菌感染患者标本160例,同时用KOH湿片法、荧光染色法和芝加哥天蓝染色法对真菌进行镜下形态学观察,并比较真菌检出率.结果 芝加哥天蓝染色法和荧光染色法对真菌的形态结构清晰易辨.芝加哥天蓝染色法、荧光染色法和KOH湿片法的检出率分别为60...     

荧光染色法、双重荧光染色法与KOH湿片法在甲真菌诊断中的比较研究

正甲真菌病由真菌感染甲板和甲床引起,以趾甲受累居多,约占皮肤科门诊足病患者的15.7%~([1])。目前皮肤科常用的实验室检查方法为氢氧化钾(KOH)直接涂片镜检和真菌培养法,但阳性率均不高。荧光染料通过与真菌细胞壁几丁质等多糖结合,在紫外光的激发下发出蓝绿色荧光。诸多研究表明荧光染色法的阳性率高于KOH湿片法~([2-5])。本研究用两种真菌荧光染色法(双重荧光染色法,荧光染色法)分别与20%KOH湿片法比较在甲真菌病检测中的阳性率、工作效率及难易程度。     

荧光染色法和KOH湿片法在诊断浅部真菌感染中的效果比较

目的探讨荧光染色法在浅部真菌镜检中的应用价值。方法收集100例皮肤科门诊疑似真菌感染的患者标本,分别运用荧光染色法和KOH湿片法对这些标本进行检测,并对检测结果进行比较。结果荧光染色法共检出56例真菌阳性,而KOH湿片法只有39例阳性。荧光染色法的阳性检出率明显高于KOH湿片法(χ2=5.79,P0.05)。结论荧光染色法是一种快速、准确的真菌检测方法,值得在皮肤科对真菌感染早期诊断的推广使用。     

荧光染色法与KOH湿片法的回顾性研究

正浅部真菌病是皮肤病中的常见病和多发病,建立一种快速而准确的检验方法尤为重要。常用的真菌检验方法有直接镜检、真菌培养、组织病理等。由于直接镜检方法简便,可快速得出检验结果,是临床最常用的检验方法之一。真菌直接镜检有多种浮载液可供选择,其中KOH使用最为广泛,其缺点是角质溶解慢,一般需要加热,加热不当易致气泡产生,且成片背景杂乱,真菌结构不易分辨和判断,易出现假阴性或漏诊;荧光染色法是一种利     

黑色素染色法在纤毛虫和鞭毛虫研究中的使用与改进

介绍并分析了前人在纤毛虫研究中使用过的黑色素染色法,得出作者自己的改进法及其复染法。报告了黑色素改进法的步骤及操作细节,并示出此法显示纤毛虫和鞭毛虫的效果。     

Visible fluorescent detection of proteins in polyacrylamide gels without staining

2,2,2-Trichloroethanol (TCE) incorporated into polyacrylamide gels before polymerization provides fluorescent visible detection of proteins in less than 5min of total processing time. The tryptophans in proteins undergo an ultraviolet light-induced reaction with trihalocompounds to produce fluorescence in the visible range so that the protein bands can be visualized on a 300-nm transilluminator. In a previous study trichloroacetic acid or chloroform was used to stain polyacrylamide gel electrophoresis (PAGE) gels for protein visualization. This study shows that placing TCE in the gel before electrophoresis can eliminate the staining step. The gel is removed from the electrophoresis apparatus and placed on a transilluminator and then the protein bands develop their fluorescence in less than 5min. In addition to being rapid this visualization method provides detection of 0.2microg of typical globular proteins, which for some proteins is slightly more sensitive than the standard Coomassie brilliant blue (CBB) method. Integral membrane proteins, which do not stain well with CBB, are visualized well with the TCE in-gel method. After TCE in-gel visualization the same gel can then be CBB stained, allowing for complementary detection of proteins. In addition, visualization with TCE in the gel is compatible with two-dimensional PAGE, native PAGE, Western blotting, and autoradiography.     

改良革兰染色法在显示石蜡切片真菌中的应用

目的建立真菌的病理切片改良革兰染色法。方法选取确诊真菌感染的组织蜡块,切若干空白片,通过苏木素-伊红染色（HE）、高碘酸-无色品红染色（PAS）、六胺银染色（GMS）、改良革兰染色后,比较改良革兰染色法的染色效果。结果改良革兰染色法的染色效果好,该法具有易操作、染色效果稳定等优点。结论改良革兰染色法能够清晰地显示出组织切片中的真菌,并且染色效果稳定,在检测组织中的真菌时可发挥重要的辅助诊断价值,具有广泛的应用前景。     

A novel,Golgi-Cox-based fluorescent staining method for visualizing full-length processes in primary rat neurons

The Golgi method, a well-known method used for staining whole dendrites and axonal trees of neurons, has been used widely for studying dendritic growth in vivo. Although detailed structural examination of neurons and their processes stained by the Golgi method has elucidated the complicated neuronal circuit, application of the method in cultured neurons has been unsuccessful to date.     

An innovative brilliant blue FCF method for fluorescent staining of fungi and bacteria

Abstract

Most natural and synthetic dyes currently used for microbial fluorescent staining are toxic or carcinogenic and are harmful to animals, humans and the environment. A food dye for microbial staining, brilliant blue FCF, was used as an alternative to lactofuchsin and lactophenol blue. Brilliant blue FCF shows pronounced microbial cell fluorescence staining of an array of pathogenic/toxigenic (Fusarium granunearum 3- and 15-acetyldeoxynivalenol chemotypes, and Escherichia coli O157:H7) and beneficial fungi and bacteria (Trichoderma harzianum and Bacillus subtilis). Brilliant blue FCF has no toxic effects on the microbes tested and is inexpensive.     

Rapid fluorescent staining of histones in sodium dodecyl sulfate-polyacrylamide gels

The increase in the fluorescence intensity of 1-anilinonaphthalene-8-sulfonate (ANS) produced by core histones is higher than that produced by very lysine-rich histones (H1 and H5). In the presence of the anionic detergent sodium dodecyl sulfate (SDS) the enhancement of ANS fluorescence caused by these two groups of histones is roughly the same, but much lower than that observed for core histones in the absence of this detergent. However, the increase of ANS fluorescence produced by histone-SDS complexes is high enough to use it for the staining of these proteins separated in SDS-polyacrylamide gels. Histone bands are stained with ANS after electrophoresis and visualized by transillumination of the gel with a uv light source. The method described in this work allows the rapid detection of less than 0.5 microgram of histone per band.     

Rapid monitoring of microbial contamination on herbal medicines by fluorescent staining method

AIMS: To apply fluorescent staining method for fast assessment of microbial quality of herbal medicines. METHODS AND RESULTS: The number of total bacteria and esterase-active bacteria on powdered traditional Chinese medicines were enumerated by fluorescent staining method using 6-carboxyfluorescein diacetate (6CFDA) and 4',6-diamidino-2-phenylindole (DAPI), and they were compared with colony-forming units (CFU). The CFU was approximately 10(3) per gram in ginseng radix, and no bacterial colonies were detected from others. However, the total bacterial number (TDC) was more than 10(7) per gram, and number of bacteria possessing esterase activity ranged from 1 to 3% of TDC. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Many bacteria in each Chinese medicine had enzyme activity and most of them could not be detected by conventional plate counting technique. Enumeration of bacterial cells on traditional Chinese medicines by fluorescent staining method requires less than 1 h. The double staining method with 6CFDA and DAPI could be applicable to rapid microbial monitoring of crude drugs.     

Blood lysate staining, a new microscopic method for diagnosis of fungemia using peripheral blood

We developed a microscopy method for the detection of fungal cells in peripheral blood, termed blood lysate staining, using an approximately 5x5 mm dotted blood lysate. This method was able to detect the emerging fungal pathogen Trichosporon asahii in murine models of systemic fungal infection and fungemia in patients quickly and at minimal cost. Pathogenic yeasts were successfully detected in 6 of 8 blood samples which were taken from feverish immunocompromised patients who were clinically suspected of having fungal infections. Fungal cells were observed as ovoid to elongated, 3x3 to 7x10 microm, and occurred singly, budding, and in short chains and clusters in a periodic acid-Schiff-stained blood smear. The yeast cells were easily distinguished from blood-cell debris by their size, shape and smooth yet rigid outline.     

Hydrothermal synthesis, characterization, and KOH activation of carbon spheres from glucose

Carbon spheres (CSs) with controllable sizes and rich in oxygen-containing groups were fabricated using a simple hydrothermal treatment of glucose. The effects of the hydrothermal parameters, including the concentration of glucose, reaction temperature, duration, and the second hydrothermal treatment were investigated. The obtained CSs were then activated using KOH for the eventual preparation of porous carbon spheres. A scanning electron microscope was used to characterize the morphology and size of the CSs. Fourier-transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to analyze the functional surface groups. N₂ adsorption–desorption isotherms were used to analyze the porous structure of the CS. The results revealed that the morphologies and size distribution of the CSs can be controlled by adjusting the experimental parameters. A hydrothermal temperature between 180 and 190 °C over 4–5 h was suitable for CS formation. Under these conditions, the size of the CS increased with the concentration of glucose. Mono-dispersed CSs with good morphologies and large numbers of oxygen-containing functional groups (primarily –OH and CO) can be obtained using a 0.3 mol/L glucose solution that is hydrothermally treated at 190 °C for 4 h. The resulting CSs sizes were about 350 nm in diameter. After a second hydrothermal treatment, the sizes of CSs grew nearly 250 nm without damage to its morphology or broadening of their size distribution. Porous CSs with perfectly spherical shapes and fully developed structures (S_BET = 1282.8 m²/g, V_micro = 0.44 cm³/g) could then be obtained via KOH activation.