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Stains And Types Of Staining Techniques Pdf

stains and types of staining techniques pdf

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Gram staining method, the most important procedure in Microbiology, was developed by Danish physician Hans Christian Gram in Gram staining is still the cornerstone of bacterial identification and taxonomic division.

By colouring otherwise transparent tissue sections, these stains allow highly trained pathologists and researchers to view, under a microscope, tissue morphology structure or to look for the presence or prevalence of particular cell types, structures or even microorganisms such as bacteria. If you have viewed this educational webinar, training or tutorial on Knowledge Pathway and would like to apply for continuing education credits with your certifying organization, please download the form to assist you in adding self-reported educational credits to your transcript. Before tissue can be stained and viewed, it must be prepared so that a very thin section, only one cell thick, can be cut and placed onto a microscope slide.

Histological Stains: A Literature Review and Case Study

The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays and immunological techniques, collectively referred to as histochemistry. Early histologists used the readily available chemicals to prepare tissues for microscopic studies; these laboratory chemicals were potassium dichromate, alcohol and the mercuric chloride to harden cellular tissues. Staining techniques used were carmine, silver nitrate, Giemsa, Trichrome Stains, Gram Stain and Hematoxylin among others. The purpose of this research was to assess past and current literature reviews, as well as case studies, with the aim of informing ways in which histological stains have been improved in the modern age. Results from the literature review has indicated that there has been an improvement in histopathology and histotechnology in stains used. There has been a rising need for efficient, accurate and less complex staining procedures.

Staining Methods – Simple Staining, Negative Staining, Gram’s Staining and Acid-Fast Staining

Show Credits Hide This Giemsa stained micrograph depicts an example of a slightly acidic slide that yielded a pink colored resultant stain. The micrograph shows malarial cells. Photo courtesy of the Public Health Image Library. Cell staining is a technique that can be used to better visualize cells and cell components under a microscope. By using different stains, one can preferentially stain certain cell components, such as a nucleus or a cell wall, or the entire cell.

Ziehl-Neelsen staining is a type of acid-fast stain, first introduced by Paul Ehrlich. Ziehl—Neelsen staining is a bacteriological stain used to identify acid-fast organisms, mainly Mycobacteria. It is named for two German doctors who modified the stain: the bacteriologist Franz Ziehl — and the pathologist Friedrich Neelsen — In anatomic pathology specimens, immunohistochemistry and modifications of Ziehl-Neelsen staining such as the Fite-Faraco method have comparable diagnostic utility. Both of them are superior to traditional Ziehl-Neelsen staining. The genus Mycobacterium is a slow growing bacteria, made up of small rods that are slightly curved or straight, and are considered to be gram positive. Some types of Mycobacteria form branches or filaments.

The microscope is a very important tool in microbiology, but there are limitations when it comes to using one to observe cells in general and bacterial cells in particular. Two of the most important concerns are resolution and contrast. Contrast, however, can be improved by either using a different type of optical system, such as phase contrast or a differential interference contrast microscope, or by staining the cells or the background with a chromogenic dye that not only adds contrast, but gives them a color as well. There are many different stains and staining procedures used in microbiology. Some involve a single stain and just a few steps, while others use multiple stains and a more complicated procedure. Before you can begin the staining procedure, the cells have to be mounted smeared and fixed onto a glass slide.

stains and types of staining techniques pdf

Ziehl–Neelsen stain

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An Introduction to Routine and Special Staining

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This technique is useful for the identification of oocysts of the coccidian species Cryptosporidium , Cystoisospora , and Cyclospora , which may be difficult to detect with routine stains such as trichrome. Concentrated sediment of fresh or formalin-preserved stool may be used. Other types of clinical specimens such as duodenal fluid, bile, pulmonary samples induced sputum, bronchial wash, biopsies may also be stained. A control slide of Cryptosporidium spp.


Because of its importance, different important stains are described in some detail. HANGING DROP TECHNIQUE The techniques employed.


It is the differential staining techniques which was first developed by Ziehl and later on modified by Neelsen. So this method is also called Ziehl-Neelsen staining techniques. Thus Ziehl-Neelsen staining techniques was developed. The main aim of this staining is to differentiate bacteria into acid fast group and non-acid fast groups. This method is used for those microorganisms which are not staining by simple or Gram staining method, particularly the member of genus Mycobacterium , are resistant and can only be visualized by acid-fast staining.

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