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Techniques for Isolation of Pure Cultures

Techniques for Isolation of Pure Cultures

Microorganisms are routinely cultured in the laboratory using various techniques to isolate, grow, and quantify them for further study. These methods rely on inoculating microorganisms into a suitable growth medium under aseptic conditions to prevent contamination. Depending on the objective, inoculation can involve direct transfer or the use of diluted bacterial suspensions as the inoculum.Streak-Plate Method for IsolationThe streak-plate method is a common technique for obtaining pure...

Need for Obtaining Pure Cultures

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Pure cultures, defined as the growth of a single microorganism species isolated from mixed populations, are fundamental tools in microbiological research and practical applications. These cultures ensure genetic and physiological uniformity, allowing researchers to study microbial traits under controlled conditions.Isolation and Maintenance of Pure CulturesObtaining a pure culture involves isolating a single microbial type from a mixed sample through techniques such as serial dilutions, streak...

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Coryneform bacteria are gram-positive, aerobic, nonmotile rods that exhibit irregular, club-shaped, or V-shaped arrangements. Their V-shape results from snapping division, where the inner cell wall layer forms the cross-wall, while the outer layer remains intact until it ruptures on one side, causing the daughter cells to bend away.The primary genera are Corynebacterium and Arthrobacter. Corynebacterium includes diverse species, ranging from saprophytes to pathogens like Corynebacterium...

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Updated: Oct 29, 2025

An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis

An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis

Published on: August 16, 2021

Culturing Mycobacteria.

Elizabeth Wallace¹, Debra Hendrickson¹, Nicholas Tolli¹

¹ATCC/BEI Resources, Manassas, VA, USA.

Methods in Molecular Biology (Clifton, N.J.)

|July 8, 2021

Summary

This summary is machine-generated.

This chapter details methods for growing Mycobacterium species, including Mycobacterium tuberculosis and Mycobacterium leprae, in laboratory settings for research and diagnostics. Protocols cover both standard cultures and specialized techniques for challenging pathogens.

Keywords:

Hypoxic culture Large-scale production of mycobacteria Mycobacterium leprae Mycobacterium tuberculosis Nontuberculous mycobacteria Normoxic culture

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Growth of Mycobacterium tuberculosis Biofilms

Growth of Mycobacterium tuberculosis Biofilms

Published on: February 15, 2012

Area of Science:

Microbiology
Mycobacterial cultivation
Infectious disease research

Background:

Early work by Robert Koch in 1882 established initial methods for growing Mycobacterium tuberculosis using coagulated bovine serum.
Continuous advancements have led to current capabilities for cultivating all known mycobacterial species.
Mycobacteria are now grown using axenic culture or in vivo passage in laboratory animals.

Purpose of the Study:

To provide conventional protocols for growing mycobacteria.
To detail methods for diagnostic purposes directly from clinical specimens.
To outline techniques for research laboratories and large-scale production.

Main Methods:

Axenic culture techniques for routine laboratory growth.
In vivo passage in laboratory animals for specific species.
Bioreactor-based large-scale production of Mycobacterium tuberculosis under normoxic and hypoxic conditions.
Cultivation of obligate intracellular pathogens like Mycobacterium leprae and Mycobacterium lepromatosis in athymic nude mice and armadillos.

Main Results:

Established protocols enable the cultivation of all known mycobacterial species.
Methods are provided for both diagnostic and research applications.
Detailed procedures for large-scale Mycobacterium tuberculosis production and cultivation of difficult intracellular pathogens are presented.

Conclusions:

Comprehensive protocols are available for growing diverse mycobacterial species.
These methods support both clinical diagnostics and scientific research.
Advanced techniques facilitate large-scale production and the study of obligate intracellular mycobacterial pathogens.