1887

An Eco-friendly, Scaled-down Gram Stain Protocol

    Author: Ruth A. Gyure1,*
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    Affiliations: 1: Western Connecticut State University, Danbury, CT 06810
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 20 May 2010
    • *Corresponding author. Mailing address: Department of Biological and Environmental Sciences, Western Connecticut State University, 181 White St., Danbury, CT 06810. Phone: (203) 837-8796. Fax: (203) 837-8769. E-mail: gyurer@wcsu.edu.
    • Copyright © 2010 American Society for Microbiology
    Source: J. Microbiol. Biol. Educ. May 2010 vol. 11 no. 1 60-61. doi:10.1128/jmbe.v1.i2.144
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    Abstract:

    Currently, flushing large volumes of Gram stain reagents into sanitary sewage systems is no longer acceptable. These chemical wastes are highly regulated and must be collected, labeled, and disposed of in a responsible manner, usually by paying a commercial service to remove them to an authorized off-site facility. Such services are costly and, as expected, costs are proportional to volume of collected waste. This “old” method of Gram staining, even if effluent is collected, generates a high volume of liquid waste which is unnecessarily diluted with additional large volumes of water from the rinsing steps.

    The purpose of using this scaled-down and eco-friendly protocol is to dramatically reduce the amount of liquid waste produced without sacrificing quality of results. This protocol is flexible, practical, and easy to implement. It does not require students to work at a bench sink, reduces user cost, and lowers environmental impact overall.

Key Concept Ranking

Gram Staining
0.71437854
Crystal Violet
0.5833333
Slides
0.4566488
0.71437854

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/content/journal/jmbe/10.1128/jmbe.v1.i2.144
2010-05-20
2017-11-23

Abstract:

Currently, flushing large volumes of Gram stain reagents into sanitary sewage systems is no longer acceptable. These chemical wastes are highly regulated and must be collected, labeled, and disposed of in a responsible manner, usually by paying a commercial service to remove them to an authorized off-site facility. Such services are costly and, as expected, costs are proportional to volume of collected waste. This “old” method of Gram staining, even if effluent is collected, generates a high volume of liquid waste which is unnecessarily diluted with additional large volumes of water from the rinsing steps.

The purpose of using this scaled-down and eco-friendly protocol is to dramatically reduce the amount of liquid waste produced without sacrificing quality of results. This protocol is flexible, practical, and easy to implement. It does not require students to work at a bench sink, reduces user cost, and lowers environmental impact overall.

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Figures

Image of FIGURE 1

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FIGURE 1

The station set-up for this protocol. Two alternative containers for collecting stain are shown. On the left is a glass culture dish (easier to keep clean); on the right is a typical disposable plastic container which is very cost-effective and easy to replace when it gets too unsightly or cracked.

Source: J. Microbiol. Biol. Educ. May 2010 vol. 11 no. 1 60-61. doi:10.1128/jmbe.v1.i2.144
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Image of FIGURE 2

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FIGURE 2

This photograph shows the amount of stain that should be “puddled” on top of the smear. Note that it should not run over the sides. Also, the clothespin helps to hold slide when moving it, but should be removed during staining so that the slide lays flat on the screen.

Source: J. Microbiol. Biol. Educ. May 2010 vol. 11 no. 1 60-61. doi:10.1128/jmbe.v1.i2.144
Download as Powerpoint
Image of FIGURE 3

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FIGURE 3

Waste from the collecting dishes must be placed in an appropriate labeled container for hazardous waste removal. At many institutions, it is acceptable to collect waste in used, washed containers such as this old bleach bottle. Alternatively, special containers may be provided by your institution. In either case, careful labeling according to state and local regulations is essential.

Source: J. Microbiol. Biol. Educ. May 2010 vol. 11 no. 1 60-61. doi:10.1128/jmbe.v1.i2.144
Download as Powerpoint

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