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Chapter 2 : Microscopy

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Microscopy, Page 1 of 2

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Abstract:

This chapter provides a general overview of infectious disease microscopy as practiced in the clinical microbiology laboratory. Additional information about the care and use of the microscope and ergonomic factors is also covered.

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2
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Figures

Image of FIGURE 1
FIGURE 1

Objective lens labeling. Objective lenses are labeled with information about the manufacturer, correction factors, numerical aperture, tube length, coverslip thickness, working distance, and expected immersion medium. Objectives without a listed aberration correction are considered achromats. Objectives without a listed immersion medium (Oil, Oel, W, Gly) are considered dry objectives and are intended for use with air between the lens and the specimen. doi:10.1128/9781555817381.ch2.f1

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2
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Image of FIGURE 2
FIGURE 2

Typical configuration for bright-field microscopy. The column of light generated by the field lens and the field diaphragm enters the bottom of the condenser and is focused on the slide by the condenser lens. The condenser diaphragm controls the angle of the light, the numerical aperture of the condenser, and the amount of contrast in the image. The working distance is the vertical distance from the top of the specimen to the leading edge of the objective lens. The semiangle of the objective aperture (θ) is used to calculate numerical aperture. Modified from reference 4. doi:10.1128/9781555817381.ch2.f2

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2
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Image of FIGURE 3
FIGURE 3

Anatomy of a typical clinical microscope with an integral camera. doi:10.1128/9781555817381.ch2.f3

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2
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Image of FIGURE 4
FIGURE 4

Dark-field illumination. The central light path interacts with the silvered dome located at the bottom of the condenser and is reflected away from the specimen. Peripheral light is reflected into the condenser, and it is reflected again by the internal condenser surfaces to produce a cone of light that is directed obliquely away from the objective. doi:10.1128/9781555817381.ch2.f4

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2
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References

/content/book/10.1128/9781555817381.mcm11.ch02
1. Douglas SD,. 1985. Microscopy, p. 813. In Lennette EH,, Balows A,, Hausler WJJr,, Shadomy HJ (ed), Manual of Clinical Microbiology, 4th ed. American Society for Microbiology, Washington, DC.
2. Abramowitz M. 2003. Microscope Basics and Beyond, vol 1. Olympus America, Inc, Melville, NY.
3. Abramowitz M. 1988. Contrast Methods in Microscopy: Transmitted Light, vol 2. Olympus America, Inc, Melville, NY.
4. Abramowitz M. 1994. Optics, a Primer. Olympus America, Inc, Melville, NY.
5. Abramowitz M. 1998. Photomicrography, a Practical Guide, vol 5. Olympus America, Inc, Melville, NY.
6. Delost MD. 1997. Introduction to Diagnostic Microbiology. Text and Workbook, p 3741. Mosby-Year Book, Inc, St Louis, MO.
7. Murphy DB,, Davidson MW. 2013. Fundamentals of Light Microscopy and Electronic Imaging. Wiley-Blackwell, Hoboken, NJ.
8. Abramowitz M. 1993. Fluorescence Microscopy: The Essentials, vol 4. Olympus America, Inc, Melville, NY.
9. Coons AH,, Creech HJ,, Jones RN. 1941. Immunological properties of an antibody containing a fluorescent group. Exp Biol Med 47:200202.
10. Coons AH,, Creech HJ,, Jones RN,, Berliner E. 1942. The demonstration of a pneumococcal antigen in tissues by use of fluorescent antibody. J Immunol 45:159170.
11. Coons AH,, Kaplan MM. 1950. Localization of antigen in tissue cells, II. Improvements in a method for the detection of antigen by means of fluorescent antibody. J Exp Med 91:113.
12. Gardner PS,, McQuillin J. 1974. Rapid Virus Diagnosis: Application of Immunofluorescence, 2nd ed. Butterworth, London, United Kingdom.
13. Goldman M. 1968. Fluorescent Antibody Methods. Academic Press, New York, NY.
14. Johnson GD,, Nogueira Araujo GM. 1981. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349350.
15. Nairn RC. 1976. Fluorescent Protein Tracing, 4th ed. Livingstone, London, United Kingdom.
16. Riggs JL,, Seiwald RJ,, Burckhalter J,, Downs CM,, Metcalf TG. 1958. Isothiocyanate compounds as fluorescent labeling agents for immune serum. Am J Pathol 34:10811524.
17. Xing Y,, Chaudry Q,, Shen C,, Kong KY,, Zhau HE,, Chung LW,, Petros JA,, O’Regan RM,, Yezhelyev MV,, Simons JW,, Wong MD,, Nie S. 2007. Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry. Nat Protoc 2:11521165.
18. Kairdolf BA,, Smith AM,, Stokes TH,, Wang MD,, Young AN,, Nie S. 2013. Semiconductor quantum dots for bioimaging and biodiagnostic applications. Ann Rev Chem 6:143162.
19. Reid T,, Baldini A,, Rand TC,, Ward DC. 1992. Simultaneous visualization of seven different DNA probes by in situ hybridization using combinatorial fluorescence and digital imaging microscopy. Proc Natl Acad Sci U S A 89:13881392.
20. Tkachuk DC,, Pinkel D,, Kuo WL,, Weier HU,, Gray JW. 1991. Clinical applications of fluorescence in situ hybridization. Genet Anal Biomol Eng 8:6774.
21. Johnson GD,, Davidson RS,, McNamee KC,, Russell G,, Goodwin D,, Holborow EJ. 1982. Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. J Immunol Methods 55:213242.
22. Giloh H,, Sedat JW. 1982. Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propylgallate. Science 217:12521255.
23. Murray RGE,. 1999. Introduction to morphology, p 520. In Gerhardt P,, Murray RGE,, Wood WA,, Kreig NR (ed), Methods for General and Molecular Bacteriology. ASM Press, Washington, DC.
24. Kofler M,, Kreczy A,, Gschwendtner A. 2002. “Occupational backache”—surface electromyography demonstrates the advantage of an ergonomic versus a standard microscope workstation. Eur J Appl Physiol 86:492497.
25. Kalavar SS,, Hunting KL. 1996. Musculoskeletal symptoms among cytotechnologists. Lab Med 27:765769.
26. Chaffin D,, Andersson G. 1991. Occupational Biomechanics. John Wiley & Sons, Inc, New York, NY.
27. Thompson SK,, Mason E,, Dukes S. 2003. Ergonomics and cytotechnologists: reported musculoskeletal discomfort. Diagn Cytopathol 29:364367.
28. Kofler M,, Kreczy A,, Gschwendtner A. 1999. Underestimated health hazard: proposal for an ergonomic microscope workstation. Lancet 354:17011702.
29. Vratney M. 1999. Considerations in microscope design to avoid cumulative trauma disorder in clinical laboratory applications. Am Clin Lab 18:8.

Tables

Generic image for table
TABLE 1

Resolving the power of selected lenses with different numerical apertures.

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2
Generic image for table
TABLE 2

Excitation and emission wavelengths of commonly used fluorochromes

Citation: Wiedbrauk D. 2015. Microscopy, p 5-14. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch2

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