STAINING TECHNIQUES IN MICROBIOLOGY & CELL BIOLOGY
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Although living microorganisms can be directly examined with the light microscope, they often must be fixed and stained to increase visibility of specific morphological features, and preserve them for future study.
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MICROBIAL STAINING
introduction
Microbial Staining - giving color to microbes. Because microbes are colorless and highly transparent structures.
Staining process in which microbes are getting color.
STAINES / DYES
Staines / dyes - organic compounds which carries either positive charges or negative charges or both .
Based on the charges
Basic stain / dyes :- stain with + ve charge .
Acidic stain / dyes - stain with -ve charge .
2. Based on function of stain :
Neutral stain / dyes - stain with both charges .
Simple staining only one dye is used differentiation among bacteria is impossible Eg . Simple Staining.
Differential staining- more than one dye is used- Differentiation among bacteria is possible- Eg. Gram's staining, Acid - fast staining.
Special staining - more than one dye used - Special structures are seen. Eg. Capsule staining , Spore staining .
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STAINING TECHNIQUES AND TYPES PROCEDURE.
This document discusses various staining techniques used in microscopy to visualize bacteria and other microscopic organisms. It describes different types of stains including simple stains that color all structures the same and differential stains that color different structures differently. Specific staining techniques are explained, including Gram staining to distinguish between Gram-positive and Gram-negative bacteria, acid-fast staining for mycobacteria, and endospore staining. The document provides details on procedures, requirements, and results for common staining methods.
Dark ground microscopy
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This document provides information about staining microorganisms. It begins by defining what a stain is and explaining that different stains can differentiate between organism types or parts. It then discusses the history of stains and the structures and mechanisms of various dyes used in staining. The remainder of the document outlines different staining techniques such as simple stains, differential stains like Gram staining, and special stains for specific structures. It provides details on reagents, principles, and methods for common staining procedures.
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bacterial staining ppt
Hereby you can get all about bacterial staining.
MICROBIAL STAINING
introduction
Microbial Staining - giving color to microbes. Because microbes are colorless and highly transparent structures.
Staining process in which microbes are getting color.
STAINES / DYES
Staines / dyes - organic compounds which carries either positive charges or negative charges or both .
Based on the charges
Basic stain / dyes :- stain with + ve charge .
Acidic stain / dyes - stain with -ve charge .
2. Based on function of stain :
Neutral stain / dyes - stain with both charges .
Simple staining only one dye is used differentiation among bacteria is impossible Eg . Simple Staining.
Differential staining- more than one dye is used- Differentiation among bacteria is possible- Eg. Gram's staining, Acid - fast staining.
Special staining - more than one dye used - Special structures are seen. Eg. Capsule staining , Spore staining .
Bright field microscopes
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CAPSULE STAINING AND METACHROMATIC STAINING.pptx
This document describes capsule staining and metachromatic staining techniques. It discusses the principle, reagents used, and procedures for each staining method. Capsule staining distinguishes capsular material from bacterial cells using negative stains like India ink or positive stains and a mordant. Metachromatic staining demonstrates granules in Corynebacterium diphtheriae using Albert's solutions, which cause the granules to appear a different color than the cells. Both techniques provide specific staining to identify important bacterial structures.
STAINING TECHNIQUES AND TYPES PROCEDURE.
This document discusses various staining techniques used in microscopy to visualize bacteria and other microscopic organisms. It describes different types of stains including simple stains that color all structures the same and differential stains that color different structures differently. Specific staining techniques are explained, including Gram staining to distinguish between Gram-positive and Gram-negative bacteria, acid-fast staining for mycobacteria, and endospore staining. The document provides details on procedures, requirements, and results for common staining methods.
Dark ground microscopy
The document discusses dark field microscopy, which uses oblique illumination to produce bright images against a dark background. It describes how dark field microscopy works by blocking axial light rays with a circular stopper, so that only scattered peripheral rays from the sample enter the objective lens to form an image. The key aspects of dark field microscopy allow it to clearly visualize unstained, transparent samples without requiring sample preparation. While simple to set up, it has some limitations such as lack of color and potential issues visualizing internal sample structures.
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This document provides information about staining microorganisms. It begins by defining what a stain is and explaining that different stains can differentiate between organism types or parts. It then discusses the history of stains and the structures and mechanisms of various dyes used in staining. The remainder of the document outlines different staining techniques such as simple stains, differential stains like Gram staining, and special stains for specific structures. It provides details on reagents, principles, and methods for common staining procedures.
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This document discusses dark field microscopy. It begins by defining dark field microscopy as a technique that produces bright objects against a dark background without using stains. This is achieved through a condenser that blocks light from entering the objective lens directly, allowing only light scattered from the specimen to pass through. Applications of dark field microscopy include viewing unstained samples like microorganisms, cells, and fibers. Advantages are its ability to examine transparent specimens, while disadvantages include sensitivity to dust or bubbles. Recent developments include using smartphone microscopy with LEDs for portable dark field imaging of nanoparticles.
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1. The document discusses various staining techniques used to visualize bacterial structures like flagella, capsules, and endospores under a microscope.
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Hans Christian Gram, a Danish physician and bacteriologist, developed the Gram staining technique in 1884 to classify bacteria. The technique involves staining a smear of bacteria with crystal violet dye, washing with iodine to form a crystal violet-iodine complex within the cell wall, then decolorizing with alcohol or acetone. Gram-positive bacteria retain the crystal violet due to their thick peptidoglycan layer, appearing dark purple under the microscope. Gram-negative bacteria lose the crystal violet during decolorization due to their thin peptidoglycan layer and outer membrane, appearing red with the counterstain. The Gram stain technique is a simple yet effective way to rapidly classify bacteria and guide
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The document discusses different types of microscopes used to view microscopic specimens. It describes light microscopes, which use lenses and visible light, including brightfield, darkfield, phase contrast, and fluorescence microscopes. It also describes electron microscopes, which use electromagnetic lenses and electrons beams to view specimens, including transmission electron microscopes that pass electrons through thin specimens, and scanning electron microscopes that scan surfaces to produce 3D images. Key aspects and uses of each microscope type are outlined.
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This document discusses various staining techniques used to visualize microorganisms under the microscope. It describes two main types of staining: positive staining, which colors the microorganisms, and negative staining, which colors the background. Specific staining methods covered include simple staining using single dyes, differential staining techniques like Gram staining and acid-fast staining, and special stains for structures such as endospores, capsules, flagella, and nuclei. Detailed procedures are provided for common staining methods along with labeled microscope images showing the results.
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2. Genetic diseases can be caused by inherited mutations, chromosomal abnormalities, or mutations in somatic cells (cancer). Inherited diseases can be due to nuclear or mitochondrial genetic mutations.
3. Examples of inherited genetic disorders and their inheritance patterns are discussed, including autosomal dominant disorders like achondroplasia and autosomal recessive disorders like thalassemia.
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Staining
This document provides protocols for several basic staining techniques used in microbiology, including simple staining, Gram staining, endospore staining, and capsule staining. Simple staining involves using a single basic dye like methylene blue or crystal violet to determine cell shape, size, and arrangement. Gram staining is a differential staining technique used to distinguish between Gram-positive and Gram-negative bacteria based on differences in their cell walls. Endospore staining uses malachite green and safranin dyes to identify bacterial endospores. Capsule staining employs India ink as a negative stain along with crystal violet to visualize uncharged polysaccharide capsules that help bacteria attach to surfaces.
403149684 stainingtechniquesgp-150203151421-conversion-gate02-pdf
The document discusses various staining techniques used in microbiology, including Gram staining, acid-fast staining, and simple staining techniques. Gram staining differentiates bacteria into gram-positive and gram-negative groups based on differences in their cell wall structure and how they retain or release crystal violet dye. Acid-fast staining uses a carbolfuchsin primary stain to identify acid-fast bacteria that resist decolorization by acid-alcohol, such as Mycobacterium tuberculosis. Simple stains like Loeffler's methylene blue and diluted carbol fuchsin are also discussed, which provide contrast but do not differentiate bacterial types.
INTRODUCTION TO MICRO LAB, STAINING TECHNIQUES & MORPHOLOGY OF BACTERIA
This PPT is helpful to understand first practical to 2nd year MBBS student.
I have added 2 video in this PPT to understand staining techniques properly.
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Staining in Microbiology
This document provides information about staining techniques used in microbiology. It discusses why staining is needed, as structural details of bacteria cannot be seen under a light microscope otherwise. It describes common staining methods like simple stains, negative stains, differential stains, and impregnation methods. Gram staining and Ziehl-Neelsen staining techniques are explained in detail, including the principles, procedures, and uses of each stain. Proper smear preparation and quality are also addressed.
Types of staining techniques
This document discusses various staining techniques used to visualize microorganisms under a light microscope. It describes simple staining which uses a single dye and differential staining techniques like Gram staining and acid-fast staining that use multiple dyes to distinguish between cell types. Gram staining distinguishes Gram-positive from Gram-negative bacteria based on differences in cell wall structure. Acid-fast staining identifies acid-fast bacteria that resist decolorization by acid-alcohol solutions, like Mycobacterium tuberculosis. Other techniques discussed include capsule staining to visualize the polysaccharide capsule of certain bacteria and wet mount preparations to directly observe motility without staining.
Presentation on Gram Staining.pptx
- Hans Christian Gram developed the Gram staining technique in 1884 while examining lung tissue from pneumonia patients under the microscope. He discovered that certain bacterial cells retained dye differently than others.
- Gram staining is a common differential staining technique that classifies bacteria as either Gram positive or Gram negative based on differences in their cell wall structure. Gram positive bacteria have a thick peptidoglycan layer that retains the primary stain, while Gram negative bacteria have a thin layer and outer membrane that washes away the primary stain.
- The Gram staining procedure involves staining a smear with crystal violet, adding a mordant, decolorizing with alcohol, and counterstaining with safranin. Gram positive bacteria appear purple/blue
Gram staining
This document provides instructions for Gram staining, a differential staining technique used to distinguish between bacteria based on their cell wall composition. It describes how Gram-positive bacteria retain the primary violet stain due to their thick peptidoglycan layer, appearing violet under the microscope, while Gram-negative bacteria's single peptidoglycan layer is decolorized by alcohol, causing them to appear pink when counterstained with safranin. The document outlines the Gram staining procedure and results, and examples of morphological appearances of common Gram-positive and Gram-negative bacteria.
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This document discusses different staining techniques used to visualize bacteria under a microscope. It describes simple staining using single dyes like methylene blue, and differential staining techniques like Gram staining and acid-fast staining. Gram staining differentiates bacteria into Gram-positive and Gram-negative groups based on their ability to retain or lose crystal violet dye. Acid-fast staining is used to identify acid-fast bacteria like Mycobacterium that appear bright red after staining. These staining methods allow clear visualization of bacterial morphology and structure.
GRAM STAINING OF BACTERIA
1. Gram staining is a differential staining technique developed by Hans Christian Gram in 1884 that is used to classify bacteria into two groups: Gram-positive and Gram-negative.
2. The key steps of Gram staining involve staining with crystal violet dye, treating with iodine, decolorizing with alcohol or acetone, and counterstaining with safranin.
3. Gram-positive bacteria retain the crystal violet dye after decolorization due to their thick peptidoglycan cell wall, while Gram-negative bacteria lose the dye due to their thinner cell wall. This allows bacteria to be classified based on their staining.
5. Staining techniques (Microbiology)
This document describes various staining techniques used to visualize bacteria under a microscope. It discusses how stains work by imparting color to bacteria and cellular structures. Specific stains mentioned include Gram stain, acid-fast stain, simple stains like Loeffler's methylene blue. The Gram stain technique colors bacteria either purple (Gram positive) or red (Gram negative) depending on their cell wall structure. The acid-fast stain uses carbolic fuchsin and retains its color after acid alcohol treatment to identify acid-fast bacteria like Mycobacterium tuberculosis. Proper staining allows clear visualization of bacterial morphology and structures.
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This document discusses various methods for identifying unknown bacterial cultures, including phenotypic, immunological, and genetic techniques. It focuses on morphological identification methods such as staining techniques like simple staining, negative staining, Gram staining, and acid-fast staining. These staining methods allow observation of bacterial size, shape, arrangement and properties to determine the taxon. Identification is important for medical, industrial, and research applications.
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This document discusses various methods used to identify unknown bacterial cultures, which is a major responsibility of microbiologists. It outlines staining techniques like Gram staining, acid-fast staining, endospore staining, and capsule staining. These techniques examine morphological characteristics of bacteria like shape, arrangement, presence of spores or capsules. The document also mentions biochemical tests that detect bacterial enzymatic activity or ability to ferment carbohydrates and produce acids/gases. Identifying pathogenic bacteria is important for medical diagnostics and food/brewing industries to prevent contamination.
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This document discusses microscopic examination techniques for intestinal parasites. Direct fecal smears can be used as a quick screening test but have limitations like small samples and false negatives. Fecal smears are examined wet or dried with stains like iodine. Concentration methods like flotation in saturated salt solutions increase parasite visibility by removing debris. Specific gravities between 1.10-1.35 are used to float different parasite eggs to the surface for examination.
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The document discusses bacterial staining techniques, specifically simple staining and Gram's staining. It begins by explaining how staining enhances contrast under the microscope since bacteria are otherwise invisible. It then describes the basic components and process of simple staining, as well as the principles and steps of Gram's staining technique. Gram's staining allows differentiation of bacteria into Gram-positive or Gram-negative categories based on differences in cell wall structure and composition. This differential staining technique is one of the most common and important in microbiology.
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Gram staining is a differential staining technique that divides bacteria into two groups - Gram positive and Gram negative - based on their ability to retain or lose crystal violet dye when treated with alcohol or acetone. Gram positive bacteria have a thick peptidoglycan cell wall that retains the crystal violet dye, appearing purple under the microscope. Gram negative bacteria have a thinner peptidoglycan layer and outer membrane that washes away the dye, leaving them stained pink by the safranin counterstain. The procedure involves staining a heat-fixed bacterial smear with crystal violet, iodine, decolorizer like alcohol, and safranin to differentiate between Gram positive and Gram negative bacteria.
Staning 1&2.pptx
This document provides information on staining techniques used in microbiology, including the purpose, procedures, and interpretations of common staining methods. It discusses:
- The purpose of staining is to increase contrast between microorganisms and the background under the microscope.
- Procedures for simple, differential, Gram, and acid-fast (Ziehl-Neelsen) staining are described in detail, including required materials and steps.
- Interpretation of results for each stain is also explained, such as Gram-positive and Gram-negative bacteria appearing different colors.
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Gram-Negative Staining Lab Report
The document discusses Gram staining techniques used to differentiate between Gram-positive and Gram-negative bacteria. Gram-positive bacteria have thicker cell walls consisting of peptidoglycan and teichoic acids, which causes them to retain the primary dye color. Gram-negative bacteria have thinner cell walls with an outer membrane, allowing the dye to be washed out by alcohol, leaving them colorless after counterstaining. The key difference observed during Gram staining is whether the bacteria retain the crystal violet dye after decolorization and counterstaining.
Bacteria Classification By Gram Staining Essay
Bizzozero staining procedure involves classifying tissues into three categories based on their mitotic activity as seen under the microscope: category I tissues with low mitotic activity, category II tissues with moderate mitotic activity, and category III tissues with high mitotic activity. The staining procedure uses proliferating cell nuclear antigen (PCNA) to label proliferating cells and support Bizzozero's 1894 tissue classification system based on mitotic index determined by examining hematoxylin and eosin stained slides under the microscope. The experiment aims to evaluate if PCNA staining agrees with Bizzozero's original tissue categorization into
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The document provides instructions for staining bacteria, including preparing smears, simple staining which uses one dye, and Gram staining which distinguishes between gram-positive and gram-negative bacteria. Proper staining techniques are described to increase contrast and visibility of bacteria under a microscope by adhering the bacteria to glass slides through drying and fixing before applying different stains.
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STAINING TECHNIQUES IN MICROBIOLOGY & CELL BIOLOGY
- 1. STAINING TECHNIQUES Presented By: Manu Bhardwaj M. Sc. Biotechnology (I Yr.) Department Of Advanced Science & Technology NIET, Nims University, Jaipur, Rajasthan, India
- 2. STAINING…? Although living microorganisms can be directly examined with the light microscope, they often must be fixed and stained to increase visibility of specific morphological features, and preserve them for future study.
- 3. STAIN TECHNIQUES SIMPLE STAIN TECHNIQUES NEGATIVE STAIN TECHNIQUES GRAM STAIN TECHNIQUES H & E STAIN TECHNIQUES
- 4. SIMPLE STAIN TECHNIQUES Place small amount of bacteria in a droplet of water on a glass slide. Air dried. Pass slide through a flame in a process called heat fixing. Which fixes the cells of slide, kill most of the micro-organism and prepare them for staining. Slide is flooded with a basic dye such as Crystal violet or Methylene blue for a minute or so. The positive charged dye is attached to the Bacterial cell which has negative charge and thus staining takes place. This technique is effective for vegetative cells. The stain does not easily penetrate spores.
- 6. NEGATIVE STAIN TECHNIQUES Opposite to the simple staining technique. Bacteria are mixed on a glass slide with an acidic dye such as Congo red or black stain Nigrosin. The mixture s smeared across the faces of slide and allow to air dry because the stain carries negative charge. It is repelled by the bacteria which also possess negative charge. The stain gathers around the cell since a chemical reaction has been take place. Because the heat fixing is avoided thus the cells appear less shrivelled or distorted. They often appear larger than stained cells and more natural.
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- 9. GRAM STAIN TECHNIQUES Prepare bacterial smear on the clean slide. Pass the slide through over the flame 2–3 times. (Heat fixing) Apply Crystal Violet (Primary stain) on smear for 1 minutes & rinse with water. Apply Gram’s iodine (Mordant) for 1 minute & wash with water. Then wash with 95% alcohol (Decolouriser) for 10-20 seconds & rinse with water. Apply Safranin (Seconday stain) for 1 minute & wash with water. Air dry, Blot dry & Observe under Microscope.
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- 11. H & E STAIN TECHNIQUES Immerse section in Hematoxylin for 1 minute. • Rinse with tap water. • Exchange tap water until the water is clear. Immerse sections in EOSIN stain for 1-2 minutes. Rinse with tap water. • Exchange tap water until the water is clear. Dehydrate in ascending alcohol solutions (50%, 70%, 80%, 95%, 100%). Clear with xylene . Mount cover slip onto a labeled glass slide with DPX.