Chapter 2: Viewing the Microbial World

Chapter 2 Viewing the Microbial World

Primary Objectives of This Chapter
Chapter 2 introduces the metric system, microscopy, and various types of microscopes. It explains the meaning of resolving power (resolution) and explains differences between simple and compound microscopes, brightfield and darkfield microscopes, and transmission and scanning electron microscopes. The information in Chapter 2 is considered essential in an introductory microbiology course.

Terms Introduced in This Chapter

After reading Chapter 2, you should be familiar with the following terms. These terms are defined in Chapter 2 and in the Glossary. Bacillus Brightfield microscope Centimeter Coccus Compound light microscope Darkfield microscope Decimeter Electron micrograph Electron microscope Empty magnification Fluorescence microscope Micrometer Microscopic Millimeter Nanometer Phase-contrast microscope Photomicrograph Resolving power Scanning electron micrograph Scanning electron microscope Simple microscope Transmission electron micrograph Transmission electron microscope

Chapter 2: Viewing the Microbial World

Review of Key Points

Microbes are said to be microscopic because some type of microscope is required to see them. Metric units are used to express the sizes of microbes. A meter (m) can be divided into 10 decimeters, 100 centimeters, 1,000 millimeters, 1 million micrometers, or 1 billion nanometers. The sizes of bacteria and protozoa are expressed in micrometers (m), whereas the sizes of viruses are expressed in nanometers (nm). A typical spherical bacterium (a coccus) is approximately 1 m in diameter. A typical rod-shaped bacterium (a bacillus) is about 1 m wide by 3 m long. Most of the viruses that cause human disease range in size from about 10 nm to 300 nm. The development of simple and compound light microscopes enabled the discovery and visualization of microorganisms. An ocular micrometer is used to measure the dimensions of objects being viewed with a compound light microscope. A simple microscope contains only one magnifying lens, whereas a compound microscope contains more than one magnifying lens. Total magnification of the compound light microscope is calculated by multiplying the magnifying power of the ocular lens by the magnifying power of the objective being used. The limiting factor of compound light microscopes is the type of illumination being used. Because visible light is used as the source of illumination, objects that are smaller than half the wavelength of visible light cannot be seen. The resolving power or resolution of an optical instrument is its ability to distinguish between two adjacent objects. The resolving power of the compound light microscope is approximately 0.2 m, which is about one-half the wavelength of visible light. When using a brightfield microscope, a person can see objects against a bright background. When using a darkfield microscope, a person sees illuminated objects against a dark background. Electron microscopes enable scientists to observe objects that are too small to be seen with a compound light microscope. For example, because they are so tiny, most viruses can only be seen using electron microscopes. Extremely small objects can be seen using electron microscopes, because electrons are used as the source of illumination. The wavelength of electrons is shorter than that of visible light. Transmission electron microscopes enable scientists to see inside of cells (i.e., to see internal details). The resolving power of a transmission electron microscope is approximately 0.2 nm,

Chapter 2: Viewing the Microbial World

which is about a million times better than the resolving power of the unaided human eye and one thousand times better than the resolving power of the compound light microscope. Using scanning electron microscopes, scientists are able to study surface details. The resolving power of the scanning electron microscope is approximately 20 nm. Photographs taken through the lens system of the compound light microscope are called photomicrographs, whereas those taken with electron microscopes are called transmission electron micrographs and scanning electron micrographs.

Increase Your Knowledge

Work through the tutorials on microscopy at: http://shs.westport.k12.ct.us/mjvl/biology/microscope/microscope.htm [This site has information on parts of the microscope, how to use the microscope, preparing wet mounts, and staining.] http://virtualurchin.stanford.edu/microtutorial.htm [Excellent site on how to use a microscope and make microscopic measurements!] Visit these web sites to learn more about metric units and conversions: http://www.austincc.edu/nrgtutor/Units/untut200.htm http://www.purplemath.com/modules/metric.htm http://convert.french-property.co.uk/ [This site has a helpful metrics conversion table just type in what you want to convert.] Visit the following excellent web site to learn about the sizes of cells and organisms: How Big Interactive at http://www.cellsalive.com/howbig.htm

Critical Thinking
1. You are planning to create a better compound light microscopeone that will enable you to see objects smaller in diameter than 0.2 m. You gather together the best lens grinders in the world and put them to work in a lens-grinding laboratory having unlimited resources. You instruct them to grind marvelous magnifying lenses and add them to an existing compound light microscope. Whats wrong with this plan? You have noticed that additional light is required as you increase magnification. Describe

2.

Chapter 2: Viewing the Microbial World

the three correct ways to increase the amount of light entering the objective lens, and one incorrect way.

Answers to the Chapter 2 Self-Assessment Exercises in the Text

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. D B A D D B A D B B

Additional Chapter 2 Self-Assessment Exercises

(Note: Dont peek at the answers before you attempt to solve these self-assessment exercises.) Matching Questions A. B. C. D. E. 10 100 1,000 1,000,000 1,000,000,000 _____ 1. _____ 2. The number of nanometers in a micrometer. The resolving power of the compound light microscope is __________ times better than the resolving power of the unaided eye. The number of micrometers in a millimeter. The resolving power of the transmission electron microscope is __________ times better than the resolving power of the scanning electron microscope. The resolving power of the transmission electron microscope is __________

_____ 3. _____ 4.

_____ 5.

Chapter 2: Viewing the Microbial World

times better than the resolving power of the unaided eye. A. B. C. D. E. 0.2 nm 20 nm 0.2 m 1 m 0.2 mm _____ 6. _____ 7. _____ 8. _____ 9. _____ 10. The width of a typical coccus. The resolving power of the unaided eye. The resolving power of the scanning electron microscope. The resolving power of the transmission electron microscope. The resolving power of the compound light microscope.

True/False Questions _____ 1. _____ 2. _____ 3. _____ 4. _____ 5. _____ 6. _____ 7. _____ 8. _____ 9. _____ 10. Anton van Leeuwenhoek is given credit for developing the first compound light microscope. The wavelength of visible light limits the size of objects that can be seen with the compound light microscope. The resolving power of compound light microscopes can be improved by adding additional magnifying lenses. A brightfield microscope can be converted to a darkfield microscope by replacing the condenser on a brightfield microscope with a darkfield condenser. Transmission electron microscopes are used to study surface features. Primary syphilis is usually diagnosed in the clinical microbiology laboratory by the use of a scanning electron microscope. A magnifying glass could be considered a simple microscope. The total magnification achieved when the oil immersion lens is used is 1,000. Fluorescence microscopy is often used in immunology laboratories. The resolving power of electron microscopes is much better than that of compound light microscopes because the wavelength of electrons is much longer

Chapter 2: Viewing the Microbial World

than that of visible light.

Answers to the Additional Chapter 2 Self-Assessment Exercises

Matching Questions 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. C C C B D D E B A C

True/False Questions 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. False (Leeuwenhoek made simple microscopes, not compound microscopes) True False (adding additional magnifying lenses to existing compound light microscopes would not improve resolving power; it is referred to as empty magnification) True False (transmission electron microscopes are used to observe internal structures; scanning electron microscopes are used to study surface features) False (primary syphilis is usually diagnosed using darkfield microscopy) True True True False (it is because the wavelength of electrons is much shorter than that of visible light)