Identification of Micro-Organisms : The identification of micro-organisms involves various techniques and methods to determine

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 Identification  of Micro-Organisms

1) Identification:

Microorganisms are considered invisible enemies of humans. The following methods are followed to identify the specific microorganism which can only be seen by microscope: smear, fixation, hanging drop preparation, straining, cultivation, animal inoculation, serological test etc.

The identification of micro-organisms involves various techniques and methods to determine the type and characteristics of micro-organisms such as bacteria, viruses, fungi, and parasites. Here are some key methods:

Morphological Characteristics

1. Shape and Size: Observe the shape and size of micro-organisms using a microscope.

2. Staining: Use staining techniques, such as Gram staining, to differentiate between types of bacteria.

Biochemical Tests :

1. Catalase Test: Determine the presence of catalase enzyme, which breaks down hydrogen peroxide.

2. Oxidase Test: Detect the presence of oxidase enzyme, which oxidizes certain substrates.

3. Sugar Fermentation: Identify micro-organisms based on their ability to ferment specific sugars.

Molecular Techniques :

1. Polymerase Chain Reaction (PCR): Amplify specific DNA sequences to identify micro-organisms.

2. DNA Sequencing: Determine the order of nucleotides in a DNA molecule to identify micro-organisms.

3. Real-Time PCR: Quantify specific DNA sequences in real-time to identify and quantify micro-organisms.

Immunological Techniques :

1. ELISA (Enzyme-Linked Immunosorbent Assay): Detect specific antibodies or antigens to identify micro-organisms.

2. Western Blot: Separate and identify proteins based on their size and immunoreactivity.

Other Techniques :

1. Mass Spectrometry: Identify micro-organisms based on their unique mass spectral fingerprints.

2. Flow Cytometry: Analyze micro-organisms based on their size, shape, and fluorescence properties.

Identification of Specific Micro-Organisms :

1. Bacteria: Use techniques such as Gram staining, biochemical tests, and molecular techniques to identify bacteria.

2. Viruses: Use techniques such as PCR, DNA sequencing, and serological tests to identify viruses.

3. Fungi: Use techniques such as microscopy, biochemical tests, and molecular techniques to identify fungi.

4. Parasites: Use techniques such as microscopy, biochemical tests, and molecular techniques to identify parasites.

(1).Microscope :

A microscope magnifies objects too small to be seen by the naked eye. It is also a tool used to visualize its structure. The science of microscopy is the science of microbiology. There are many types of microscopes. Three of them are very important.

1. Compound microscope helps in routine bacterial tests.

2. Microscope is used to detect spirochetes. E.g. Trepenoma pallidum

3. The electron microscope is a very powerful microscope. It magnifies the object by 1,00,000 times. It is used to detect viruses.

A microscope is an essential tool in various fields, including biology, medicine, and materials science. Here's an overview:

Types of Microscopes :

1. Light Microscope (LM): Uses visible light to illuminate samples.

2. Electron Microscope (EM): Uses a beam of electrons to produce high-resolution images.

3. Fluorescence Microscope: Uses fluorescent dyes to visualize specific structures or molecules.

4. Confocal Microscope: Uses laser light to produce high-resolution, 3D images.

5. Scanning Probe Microscope: Uses a physical probe to scan the surface of samples.

Components of a Microscope :

1. Eyepiece: The lens through which the user looks.

2. Objective Lenses: The lenses that collect and focus light from the sample.

3. Stage: The platform that holds the sample.

4. Illuminator: The light source that illuminates the sample.

5. Focus Knobs: The controls that adjust the focus of the microscope.

Microscope Techniques :

1. Brightfield Microscopy: Uses transmitted light to produce a dark image on a bright background.

2. Darkfield Microscopy: Uses scattered light to produce a bright image on a dark background.

3. Phase Contrast Microscopy: Uses differences in refractive index to produce contrast.

4. Differential Interference Contrast (DIC) Microscopy: Uses differences in refractive index to produce contrast.

Applications of Microscopy :

1. Biological Research: Studying cells, tissues, and microorganisms.

2. Medical Diagnosis: Diagnosing diseases, such as cancer and infectious diseases.

3. Materials Science: Studying the structure and properties of materials.

4. Quality Control: Inspecting products for defects and contaminants.

(2). Smearing:

A drop of the sample to be tested is placed in the center of the microplate. A drop of distilled water (water required only for solid samples e.g. faeces) is taken through a cleaned (sterilized) loop and left on the sample, spreading the sample evenly on the microplate. This type of test is a direct microscopic test.

(3). Fixing:

The coating is applied by exposing the microplate to a flame. Flaming is done by showing the microplate three or four times in the flame. Fixation is done using chemicals such as formalin, methyl alcohol and mercuric chloride.

(4). Hanging Drop Preparation:

This method is often used to determine whether bacteria are motile or nonmotile. The preparation helps to study living organisms.

(5).ELISA:

Human immunodeficiency virus (HIV) is detected by ELISA and Western blot tests.

2) Types of Samples Collected (Laboratory Techniques):

(i) Swabs :

It is usually collected in a sterile test tube. Care should be taken to prevent contamination of the sample.

(ii). Throat Swab Artificial Cultivation :

View the throat in good light. Throat swab cultures should be taken later using a tongue depressor. The product should only be applied to the infected area.

(iii) Cowardly (cold) :

It should be collected in sterilized wide-mouthed containers. It should be collected directly after a cough and sent to the laboratory immediately.

(iv). Small water :

A urine sample is subjected to chemical and microscopic tests. should be collected in a clean container or test tube. But for culture it must be collected in a sterile test tube.

(v). Faeces :

               

A fresh stool should be collected for bacterial testing.

(vi). Blood:

It should be collected in a sterilized container. The nurse should assist when collecting other specimens, such as cerebrospinal fluid. and other, preventive measures should be taken during medical assistance to prevent the spread of germs from the specimen and to avoid being infected by the germs themselves.