JoVE Science Education

Environmental Microbiology

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English

Aseptic Technique in Environmental Science

Przegląd

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - The University of Arizona
Demonstrating Author: Luisa Ikner

Aseptic technique is a fundamental skill widely practiced in the field of environmental microbiology that requires a balance of mindfulness and practice in the laboratory. Proper use of this technique reduces the likelihood of bacterial or fungal contamination of reagents, culture media, and environmental samples. Aseptic technique is also vital to ensure data integrity and maintain the purity of culture libraries that may be comprised of very rare and difficult to culture isolates. Sources of contamination in the laboratory environment include airborne microorganisms (including those adhering to dust and lint particles), microbes present on the laboratory bench workspace or on unsterilized glassware or equipment, and microbes transferred from the body and hair of the researcher. The use of aseptic technique is also a safety measure that lowers the potential for the transmission of microorganisms to researchers, which is particularly important when working with pathogens.

Procedura

1. Preparation for Aseptic Work

  1. Obtain and apply the following PPE items: lab coat, latex or nitrile gloves (free from tears or holes), and safety goggles (Figure 1). For safety in the event of using an open flame, tie back long hair.
    Figure 1
    Figure 1: PPE: A lab coat, latex gloves, and safety goggles.
  2. A second important aspect of aseptic technique is the proper sterilization and storage of med

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Wyniki

The outcome of the procedure demonstrates proper aseptic technique and poor aseptic technique. Figure 7 illustrates the contamination that can arise from poor aseptic technique when pouring agarose plates (top plate: sterile medium; bottom plates: contaminated media).

Figure 7
Figure 7: Contamination that can arise from poor aseptic technique when pouring agarose plate

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Wniosek i Podsumowanie

Other than using Bunsen burners, aseptic working environments can also be maintained in specialized workstations known as laminar flow hoods, which use directed airflow and filters to maintain sterility.

Proper use of aseptic technique is vital for environmental microbiologists when sampling in the field and in the laboratory when working with media, reagents, and cultured isolates.  Poor aseptic technique in the field can result in the transfer of microorganisms from the technician to cr

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Przejdź do...

0:00

Overview

1:10

Principles of Aseptic Technique

3:02

Preparation for Aseptic Work

5:10

Aseptic Transfer of Bacteria Between Liquid Media and Petri Plates

8:13

Applications

10:20

Summary

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