微生物实验中常用细菌染色方法的改良

细菌染色方法是微生物实验中观察细菌形态结构以及掌握细菌代谢、繁殖等情况的常用普及技术。微生物实验中常用的细菌染色法包括其革兰染色法、抗酸染色法等。为进一步提高实验中细菌形态的观测质量,本文就对两种常用细菌染色法进行了改良,据实验结果显示,其改进后方法操作更加简单,更有效起到细菌染色作用及观测作用。     

HACCP体系在细菌形态学检测之革兰氏染色中的应用

细菌形态学检测方法是细菌检验中极为重要的鉴定手段之一,革兰氏染色法是细菌中最常见的一种鉴别染色法,是细菌分类鉴定中的重要标志。革兰氏染色法常因各种因素造成误差,造成反应结果的不稳定。因此,我们需要依靠有效的质量控制措施来消除影响染色结果的各种误差。本文将HACCP体系应用到革兰氏染色中,确定革兰氏染色的关键控制点和关键限值,规范了试验操作,以便更好地保证染色结果的准确性。     

神经髓鞘染色新方法的建立及其原理探讨

在神经病理诊断和研究工作中,髓鞘染色是一种常用的方法。任何因素的神经纤维损伤,均可导致髓鞘的变性、崩解或脱失。普通染色中髓鞘不易着色,在正常或病理情况下均需要特殊染色法来观察髓鞘的损害程度。常规髓鞘特殊染色多采用传统的苏木素（如Wei1氏和Loyez氏法）及锇酸染色法。     

三种细菌鞭毛染色法的比较

三种细菌鞭毛染色法的比较济宁医学院２７２１１３刘昌平,陈恩华鞭毛是某些细菌的一种特殊结构,必须通过特殊的染色法,将纤细的鞭毛增粗染色后,才能在一般光学显微镜下观察到,但由于影响因素较多,直接鞭毛染色往往难获成功,为了获得一种实验教学较为满意的染色法,...     

一种新的细菌荚膜染色方法

一种新的细菌荚膜染色方法孙迅,王宜磊,朱陶（山东省菏泽高等师范专科学校生物系,２７４０１５）本文推荐一种用淀粉作为背景衬托基质的细菌荚膜染色法。该法能够较好地适用于实验教学常用菌株如圆褐固氮菌等的荚膜染色。荚膜染色无论在实验教学还是在细菌学鉴定等方面...     

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。     

免疫组化染色和特殊染色对显示幽门螺杆菌的效果影响对比

目的探讨分析免疫组化染色和特殊染色对显示幽门螺杆菌的效果影响对比。方法选取2013年9月-2014年9月在我院的胃黏膜活检的组织标本共120例,将进行三种染色方法的检测,分为三组。Giemsa组:使用Giemsa染色法进行染色;中红组:使用中性红染色法进行染色;免疫组:使用免疫组织化学法进行染色,比较三组染色的特点。结果结果显示,染色阳性率:Giemsa组为45.0%,中红组为37.5%,免疫组为62.5%。结论可见免疫组化染色阳性率最高,具有比较好的检测显示效果,因此随着免疫组织化学技术的发展,未来会给检测幽门螺杆菌带来更大的发展和进步。     

芽孢染色法

芽孢染色法芽孢是细菌细胞的特殊结构，是产生芽孢菌的重要特性．芽孢染色在细菌鉴定上有重要意义。因芽孢壁厚透性差．故不易染色。通常用孔雀绿长时间加热，才能使芽孢着色。这种方法往往很难将所有的芽孢着色。现介绍一种新的染色法。材料：在肉汤琼脂培养基上培养２４...     

金胺O荧光染色在石蜡组织切片麻风病诊断中的应用

目的为了验证金胺O荧光染色法应用于石蜡组织切片麻风杆菌检测的可行性。方法用金胺O荧光法对6例确诊为麻风病的病理组织切片进行染色,并与抗酸染色结果进行对比。结果荧光染色法6例结果均为阳性,在暗背景下麻风杆菌显示明亮淡绿色荧光;在菌量较少荧光染色片中寻找单根麻风杆菌,较抗酸染色片更为容易。结论金胺O荧光染色法可用于石蜡组织切片麻风病的诊断,麻风杆菌单根散在时比抗酸染色法有一定优势。     

荚膜染色纸片的制备和应用

本文报告细菌荚膜染色纸片的制备和应用结果。与常用的墨汁法相比较,证明了用荚膜染色纸片进行细菌荚膜染色的可能性。本法操作简单,纸片携带方便.染色效果良好,有推广应用价值。     

革兰氏染色三步法与质量控制

革兰氏染色(Gram stain),是细菌学中一个经常使用和十分重要的方法,自从1884年微生物学家Gram氏发明著名的革兰氏染色法以后,100多年来虽然经过后来学者的几次改进,但都仍然沿用着Gram氏原来的四步法,基本原理也没有改变。最近Allen氏对Ziehl-Neelsen抗酸菌染色法的改进,是一个良好的启示,使我们开始了革兰氏染色三步法的研究并取得了成功。现将我们建立的革兰氏染色三步法与质量控制报告如下。 1 材料和方法 1.1 结晶紫染色液 甲液:结晶紫2g;95％乙醇20ml。 乙液:草酸铵0.8g;蒸馏水80ml。 甲乙二液先分别溶解,然后混合在一起,过滤除去残渣后装入滴瓶中备用。     

Adaptations of Goldner's Masson Trichrome Stain for the Study of Undecalcified Plastic Embedded Bone

Specialized adaptations for application of Goldner's Masson trichrome stain to plastic embedded undecalcified bone specimens are presented. This stain can be used successfully on methyl-glycol methacrylate, glycol methacrylate and Spurr embedded bones. The stain affords the advantage of good cellular staining due to the hematoxylin component with concomitant sharp discrimination of mature bone matrix which stains green, immature new bone matrix which stains red, and calcified cartilage which stains very pale green. Use of red filters during photomicrography aids in bone-osteoid discrimination in black and white photographs.     

Rapid,in situ detection of Agrobacterium tumefaciens attachment to leaf tissue

Attachment of the plant pathogen Agrobacterium tumefaciens to host plant cells is an early and necessary step in plant transformation and agroinfiltration processes. However, bacterial attachment behavior is not well understood in complex plant tissues. Here we developed an imaging‐based method to observe and quantify A. tumefaciens attached to leaf tissue in situ. Fluorescent labeling of bacteria with nucleic acid, protein, and vital dyes was investigated as a rapid alternative to generating recombinant strains expressing fluorescent proteins. Syto 16 green fluorescent nucleic acid stain was found to yield the greatest signal intensity in stained bacteria without affecting viability or infectivity. Stained bacteria retained the stain and were detectable over 72 h. To demonstrate in situ detection of attached bacteria, confocal fluorescent microscopy was used to image A. tumefaciens in sections of lettuce leaf tissue following vacuum‐infiltration with labeled bacteria. Bacterial signals were associated with plant cell surfaces, suggesting detection of bacteria attached to plant cells. Bacterial attachment to specific leaf tissues was in agreement with known leaf tissue competencies for transformation with Agrobacterium. Levels of bacteria attached to leaf cells were quantified over time post‐infiltration. Signals from stained bacteria were stable over the first 24 h following infiltration but decreased in intensity as bacteria multiplied in planta. Nucleic acid staining of A. tumefaciens followed by confocal microscopy of infected leaf tissue offers a rapid, in situ method for evaluating attachment of A. tumefaciens' to plant expression hosts and a tool to facilitate management of transient expression processes via agroinfiltration. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

微量蛋白检测中不同银染方法的比较与分析

随着生物化学技术的不断发展,作为检测SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)中微量蛋白的银染方法也在不断改进和发展.采用4种不同的银染方法检测不同含量的牛血清白蛋白,结果显示单纯的银染过程中如果使用戊二醛固定会使蛋白检出更快速灵敏,而结合考马斯亮蓝的复合银染则较单纯银染灵敏度提高了5～7个数量级.     

A New Stain for Pollen Fertility Studies

The juice from the berries of Cocculus hirsutum was extracted and used for pollen fertility studies in various crops. Two stains were prepared: P. H. Ramanjini (PHR) stain and modified PHR stain. The modified PHR stain contains lactic acid and produces the best staining differentiation. The intensity of the staining was dependent on the thickness of the pollen cell walls, hence PHR stain is recommended for thick walled pollen grains and the modified PHR stain for pollen with relatively thin walls. The preparation of both the stains are very simple, quick and inexpensive.     

A high-affinity reversible protein stain for Western blots

We describe a reversible staining technique, using MemCode, a reversible protein stain by which proteins can be visualized on nitrocellulose and polyvinylidine fluoride (PVDF) membranes without being permanently fixed to the membrane itself. This allows subsequent immunoblot analysis of the proteins to be performed. The procedure is applicable only to protein blots on nitrocellulose and PVDF membranes. MemCode is a reversible protein stain composed of copper as a part of an organic complex that interacts noncovalently with proteins. MemCode shows rapid protein staining, taking 30s to 1 min for completion. The method is simple and utilizes convenient application conditions that are compatible with the matrix materials and the protein. The stain is more sensitive than any previously described dye-based universal protein staining system. The turquoise-blue-stained protein bands do not fade with time and are easy to photograph compared to those stained with Ponceau S. Absorbance in the blue region of the spectrum offers good properties for photo documentation and avoids interference from common biological chromophores. The stain on the protein is easily reversible in 2 min for nitrocellulose membrane and in 10 min for PVDF membrane with MemCode stain eraser. The stain is compatible with general Western blot detection systems, and membrane treatment with MemCode stain does not interfere with conventional chemiluminescent or chromogenic detection using horseradish peroxide and alkaline phosphatase substrates. The stain is also compatible with N-terminal sequence analysis of proteins.     

Advanced negative detection method comparable to silver stain for SDS-PAGE separated proteins detection

In order to achieve an easy, rapid and sensitive protocol to detect proteins in polyacrylamide gel, an advanced negative detection method comparable to silver stain is described. When a gel was incubated with Phloxine B and followed by the development in acidic solution, the zones where forming protein-dye complex were selectively transparent, unlike opaque gel background. Within 50 min after electrophoresis, down to 0.1–0.4 ng of gel-separated proteins (similar with silver stain) could be observed, without labor-intensive and time-consuming procedure. Comparing with the most common negative stain method, Imidazole-zinc stain, Phloxine B stain has been shown higher sensitivity and distinct contrast between the transparent protein bands/spots and opaque background than those; furthermore, it is no longer necessary to concern about retention time of observation. This technique may provide a sensitive and practical choice for proteomics researches.     

Localization of methylene blue paramolybdate in vitally stained nerves

Methylene blue taken up by living neurons can be preserved for electron microscopy in a fixative containing osmium tetroxide and ammonium paramolybdate at pH 5.2. Paramolybdate is the buffer, precipitating agent and main osmotic ingredient; it does not function as an electron stain unless methylene blue is present. The low pH keeps the dye/paramolybdate complex from dissolving. Neither the low pH nor drastic dehydration from water to absolute ethanol harm the tissue. The staining mechanism involves cationic methylene blue associating with anionic structures such as microtubules and neurofilaments in the living cell; during fixation paramolybdate forms a precipitate with the dye at the staining sites. This fixative does not preserve microtubules unless they are first vitally stained.     

Tinctorial Properties of Spherical Bodies in Broth Cultures of Nocardia asteroides GUH-2

In yeast extract-supplemented brain heart infusion (BHI) broth cultures of Nocardia asteroides GUH-2, many spherical bodies (SBs) were frequently seen nearby filamentous cells. They showed no Gram-positivity when Gram stain was applied. When acridine orange stain was applied, many of them showed different green fluorescence from bright orange fluorescence of the filamentous nocardiae under ultraviolet light. Their acid-fastness appeared to depend on the presence of paraffin. Using the polymerase chain reaction (PCR) method, 16S rRNA genes were detected in SB-containing broth cultures inoculated with culture filtrates from broth cultures of the strain and identical to that of N. asteroides. These results suggest that SBs are cell wall-defective (CWD) forms which result from the spontaneous mutation of N. asteroides GUH-2.     

Evaluation of stain removal and inhibition properties of eight denture cleansers: an in vitro study

Gerodontology 2012; doi: 10.1111/j.1741‐2358.2011.00522.x
Evaluation of stain removal and inhibition properties of eight denture cleansers: an in vitro study Objectives: To determine the ability of eight denture cleansers to remove and inhibit tea‐stain build‐up on acrylic resin. Materials and methods: In the stain removal study, Perspex^® (cast heat polymerised resin) specimens previously soaked in saliva were stained using multiple exposures of chlorhexidine and tea solutions. Specimens were exposed for 1 min to one of the eight denture cleansers for five cycles, washed and dried and their optical density read on a uv/vis spectrophotometer at 295 nm. In the stain inhibition study, clear specimens were exposed to saliva followed by cleansers then tea solution, for five cycles. The build‐up of stain at each cycle was measured, and differences in optical densities from baseline were calculated. Results: All denture cleansers were significantly more effective than water in removing stain (p < 0.05). There were significant differences in cleaning ability between cleansers (p < 0.001), Dentural^® and Kleenite^® were particularly effective. The stain inhibition experiment showed that most cleansers were significantly more effective than water in inhibiting stain (p < 0.05). There were significant differences in inhibition ability between cleansers (p < 0.01). Kleenite^® and Equate were particularly effective. Conclusions: All denture cleansers had a capacity to remove stain and most had an inhibitory effect on staining. Kleenite^® was particularly effective in controlling stain formation.