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Chapter 3.2 : Staining Procedures

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Staining Procedures, Page 1 of 2

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Abstract:

The Gram stain has many uses: principally, it classifies bacteria on the basis of their cell wall structure and allows observation of their size and cellular morphology as well. It can also be utilized to assess the quality of clinical specimens and as a critical test for rapid and presumptive diagnosis of infective agents directly from specimens ( ). The stain was originally developed by Christian Gram in 1884 ( ). The modification currently used for general bacteriology was developed by Hucker in 1921, providing improved consistency of decolorization and better differentiation of organisms ( ). Kopeloff’s modification, which employs a basic fuchsin (or carbol fuchsin) counterstain ( ), has particular utility for staining anaerobes and weakly staining Gram-negative organisms (e.g., , and ) ( ). Consequently, many laboratories use these counterstains routinely, especially for direct smears of clinical material.

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
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Figures

Image of Figure 3.2.1–1
Figure 3.2.1–1

Thin-smear preparation.

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
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Image of Figure 3.2.1–2
Figure 3.2.1–2

Touch preparation of smear.

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
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Image of Figure 3.2.1–3
Figure 3.2.1–3

Typical Gram stain morphologies of Gram-positive and Gram-variable genera.

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
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Image of Figure 3.2.1–4
Figure 3.2.1–4

Typical Gram stain morphologies of Gram-negative genera.

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
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References

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Tables

Generic image for table
Table 3.2.1–1

Gram stain modifications, recommended reagents, timing, and uses

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
Generic image for table
Table 3.2.1–2

Gram-positive organisms found in direct smears from clinical sources

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
Generic image for table
Table 3.2.1–3

Gram-negative organisms seen in direct smears from clinical sources

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
Generic image for table
Table 3.2.1–4

Common descriptions of bacterial Gram staining characteristics

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
Generic image for table
Table 3.2.1–5

Reporting Gram stain results

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
Generic image for table
Table 3.2.1–A1

Standardized scoring method for evaluation of Gram stains for BV

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2
Generic image for table
Table 3.2.3–1

Outline for reporting wet mount results from different specimens

Citation: Chan W. 2016. Staining Procedures, p 3.2.1.1-3.2.3.6. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.2

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