Microbial Culture Media

Microbial Culture Media The micro-organism requires a base medium for their growth and it is called culture media. Nutrient media contains the substances that are used by organism synthesis of food, growth and survival. It fulfils all the nutritional requirements of the micro-organism. The nutrient media consist of water as the important ingredient plus organic … Read moreMicrobial Culture Media

Staining of Fungus by using Lactophenol cotton blue

Staining of Fungus by using Lactophenol cotton blue Aim To stain fungal cells by using Lactophenol cotton blue. Approach Fungi are eukaryotic organism and they are classified into two main groups that is yeast and molds.Its cell wall is made up of chitin.Fungal cells have both macroscopic as well as microscopic structure. This microorganism is … Read moreStaining of Fungus by using Lactophenol cotton blue

Rickettsia Staining by using Gimenez Method

Rickettsia Staining by using Gimenez Method Aim To stain Rickettsia by using Gimenez staining method. Approach Rickettsia is a non-motile, Gram-negative,non-spore forming, pleomorphic bacteria. It is pleomorphic because it shows different shapes of bacteria that is cocci, rod and spiral shape. Rickettsia name was given by scientist Howard Ricketts. It is an obligate intracellular parasite … Read moreRickettsia Staining by using Gimenez Method

Nuclear Staining by Giemsa’s Method and its Details

Nuclear Staining by Giemsa’s Method and its Details Aim To stain bacteria nuclear material by Giemsa’s staining technique. Approach DNA is a principle genetic material of all micro-organisms except some viruses. Bacterial cell lacks a well-organised nucleus. It is the very essential structure of a living cell. Nucleus performs important functions like growth, metabolic activities, … Read moreNuclear Staining by Giemsa’s Method and its Details

Flagella Staining by Loeffler’s Method and its Details

Flagella Staining by Loeffler’s Method and its Details

Flagella is the most important locomotory organ of a bacterial cell and for studying this structure it is important to stain the flagella.So here we have another staining technique used for staining of flagella that is Loffler’s staining technique.

There are some other staining techniques that are also used for staining of flagella that are Bailey’s method,Gray’s method, Casares-Gil’s method and Leifson’s method.

Here in this article we are going to study  Loffler’s method in detail.

Aim

To stain flagella of a cell by using Loffler’s method.

Requirement

  1. Flagellated cell culture.
  2. Loeffler’s mordant.
  3. Loeffler’s stain.
  4. Distill water.
  5. Chromic acid.

Procedure

  1. The slides are cleaned by boiling them in chromic acid solution.
  2. The smear is prepared as given in the flow chart below.
  3. Further after smear preparation the slide is treated with Loeffler’s mordant and heated it till steam appears approximately it takes 3 minutes.
  4. After that slide is gently washed by distill water.
  5. Further slide is treated with Loeffler’s stain and heated till steam appears.
  6. Finally slide is washed with water,air dried and observe under oil immersion.

Read moreFlagella Staining by Loeffler’s Method and its Details

Spirochete Staining by Becker’s Method

Spirochete Staining by Becker’s Method

  • Spirochete are spiral shape bacteria .Becker’s staining is another special staining used to stain spirochete.
  • Other Spirochete staining techniques are Rue’s method and Levaditi’s method.

Aim

To stain spirochete by using Becker’s method.

Requirement

  1. Tartar sample.
  2. Fontana’s fixative.
  3. Fontana’s mordant.
  4. Becker’s stain.

Procedure

  1. First of all a sterile pin is used to take a sample of tartar from teeth.
  2. The tartar sample is mixed and homogenized in sterile distill water and further used as a suspension.
  3. A drop of suspension is taken on slide ,smear is prepared,air dried and  fixed by using chemical fixative that is Fontana’s fixative for 30 seconds for 2 times after interval of 1 minute.
  4. The slide is further washed with water,air dried and treated with Fontana’s mordant for 5 minutes.
  5. After 5 minutes the mordant is drained off and slide is given a water wash treatment.
  6. Further the slide is treated with Becker’s stain for 3 to 5 minutes.
  7. Finally slide is washed with water,air dried and observe under oil immersion.

Read moreSpirochete Staining by Becker’s Method

Spirochete Staining by Fontana’s Technique

Spirochete Staining by Fontana’s Technique

Introduction

  • Spirochete are spiral shaped Gram negative bacteria.
  • They are unique in morphology and locomotion.
  • The spirochete is spiral in shape so its size is measured by measuring its length and width.
  • Generally, it is 5 to 500 µm in length and 0.1 to 3 µm in width.
  • Spirochete has endo-flagella that runs through periplasm.
  • The endo-flagella maybe 2 to 100 on the number and these flagella are wound around the helical cells in between protoplast and outer membrane.
  • These spirochetes are extremely delicate organism so some special technique is used to stain them.

There are two technique to stain these spiral shape organism.

  1. Fontana’s staining technique.
  2. Becker’s staining technique.

Here we are going to study Fontana’s staining technique in detail.

Fontana’s staining technique

Aim

To stain spirochete by using Fontana’s staining technique.

Requirement

  1. Fontana’s fixative.
  2. Fontana’s mordant.
  3. Fontana’s stain.
  4. 95% Ethanol.
  5. Tartar sample.

Read moreSpirochete Staining by Fontana’s Technique

Rules and Safety Measures in Microbiological Laboratory.

Rules and Safety Measures in Microbiological Laboratory It is very important to follows rules of microbiological laboratory in safety point of view. If these important rules are not followed a student,teacher, or a laboratory worker can face Biohazards, Chemical hazards as well as Physical hazards. The safety rules are as follows Whenever a person enters … Read moreRules and Safety Measures in Microbiological Laboratory.

Nomenclature of Bacterial Cell

Nomenclature of Bacterial Cell

In this world, there are millions of microorganisms.These microorganism are present everywhere and each organism has its unique and special features.There is a huge diversity in morphological, physical, and chemical characters of these organism.So due to such a huge diversity it is very important to name and classify these organism.Nomenclature and classification of these micro-organism can avoid confusion and misunderstanding .Due to huge diversity of these organism they are named into different names which leads to confusion.So naming these organism in a systematic and logical way is very important.

These organism are named in two ways.

  1. Common Name
  2. Scientific Name

Read moreNomenclature of Bacterial Cell

Flagella Staining Technique by Liefson’s Method

Flagella Staining Technique by Liefson’s Method

Introduction

  • Bacteria have two types of locomotory organs and that are Flagella and pili.
  • Here we are studying about flagella staining.
  • Flagella is a thin,hair like structure made up protein called as flagellin.
  • It size ranges from 20 µ to 200 µ in length.
  • Flagella is one of the most important locomotory organ .It is mainly made up of three parts- 1) Basal body 2) Filament 3) Hook.
  • Flagella is generally present in rod shape bacteria and very few cocci shape bacteria posses flagella.
  • As flagella are very thin and hair like they cannot be easily observed under microscope.
  • So a special technique is design to increase thickness of flagella as well as stain it.
  • Due to these technique we can observe structure of flagella easily under microscope.

Please check out the below video for Flagella staining by Liefson’s method.

Aim

To Stain Flagella of cell by using Leifson’s method

Requirement

  1. Flagellated cell culture slant.
  2. Leifson’s stain.
  3. 1 % Methylene blue.
  4. Distilled water.

Procedure

  • First of all take two hours old flagellated cell culture slant and add two to three drops of sterile distill water in the slant with the help of sterile pipette.
  • Note that the distill water is added slowly without disturbing the growth of cells.
  • After addition of distill water incubated the slant for 20 minutes.
  • Then take a drop of suspension from the slant and place the drop on a clean slide which is kept in slanting position.
  • The drop should flow slowly from one end of slide to other end to avoid folding of flagella on cell.
  • Allow smear to air dry here we don’t use heat fixation treatment .
  • After air drying the slide is flooded with Leifson’s stain till a thin film of shinny surface appear.
  • After this give a gentle stream of water wash treatment to a slide.
  • Now treat the slide with 1 % methylene blue treatment for 1 minute.
  • Give the slide water wash treatment ,air dry and observe under oil immersion lens.

Read moreFlagella Staining Technique by Liefson’s Method

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