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Flashcards in Chapter 4 Deck (92):
1

A unit of measurement is?

Smaller than the object being measured

2

Scientists use what type of measurements?

Metric units

3

The metric system is a?

Decimal system

4

Unit of length in the metric system is?

Meter (m)

5

One tenth of a meter is?

decimeter (dm)

6

One one-hundreth of a meter is a?

Centimeter (cm)

7

One tenth of a centimeter is?

millimeter (mm)

8

In the metric system, we continue to divide by multiples of?

10

9

One-thousandth of a millimeter is?

A micrometer which is small enough to be useful in measuring cells

10

Unit used to measure the smallest cellular organelles and viruses

nanometer (nm) which is one-billionth of a meter

11

Microscopy refers to?

The use of light or electrons to magnify objects

12

Science of microscopy began when

Leeuwenhoek used primitive microscopes to observe & report the existence of microorganisms

13

General principle involved in both light and electron microscopy include

The wavelength of radiation, the magnification of an image, the resolving power of the instrument, and contrast in the specimen

14

Beams of radiation may be referred to as either

Rays or waves

15

rays and waves (various forms of radiation) differ in?

Wavelength, the distance between two corresponding parts of a wave

16

The human eye discriminates among different wave lengths of visible light & sends patterns of?

Nerve impulses to the brain, which interprets the impulses as different colors. For example we see wavelengths of 400nm as violet, & 650 nm as red. White light (composed of many colors (wavelengths), has an average wavelength of 550 nm

17

Electrons are negatively charged particles that orbit the nuclei of atoms. Besides being particulate, moving electrons act as?

Waves, with wavelengths dependent on the voltage of an electron beam. For example, the wavelength of electrons at 1,000,000 V is 0.001 nm. Radiation of smaller wavelengths results in enhanced microscopy

18

Magnification is the?

Apparent increase in size of an object. It is indicated by the number and "X", which reads "times." For example, 16,000X is 16,000 times

19

Magnification results when a?

Beam of radiation refracts (bends) as it passes through a lens. Curved glass lens refract light, and magnetic fields refract electron beams

20

A lens refracts light because?

The lens is optically dense compared to the surrounding medium (such as air); that is, light travels more slowly through the lens than through air

21

Because of its curvature, a lens refracts light rays that pass through its periphery more than light rays that pass through its?

Center, so that the lens focuses light rays on a focal point

22

The degree to which an image in enlarged depends on?

The thickness of the lens, its curvature, & the speed of light through its substance

23

The properties that determine the clarity of an image of a microscope, are?

Resolution and contrast

24

Resolution is?

Also called the resolving power, is the ability to distinguish objects that are close together

25

Modern microscopes have ________ resolution than Leeuwenhoek's; they can distinguish objects as close together as 0.2 micrometers

Fivefold

26

Ability of the lens to gather light

Numerical aperture

27

Contrast refers to

Differences in intensity between two objects or between an object and its background. Important in determining resolution

28

What does phase mean?

All the waves crests and troughs are aligned

29

Bright-field microscopes (most common microscopes)

The background (or field) is illuminated

30

Dark-field microscopes

The specimen is made to appear light against a dark background

31

Phase microscopes use the

Alignment or misalignment of light waves to achieve the desired contrast between a living specimen & its background

32

Fluorescent microscopes use

Invisible ultraviolet light to cause specimens to radiate visible light

33

Microscopes that use lasers to illuminate fluorescent chemicals in a thin plane of a specimen are called

Confocal microscopes

34

Staining increases?

Contrast and resolution

35

What type of microscopy requires that specimens be treated with stains or coatings to enhance contrast?

Electron microscopy

36

Thin film of organisms on a slide is called a?

Smear

37

If the organisms are growing on a solid surface, such as an agar plate, then they are?

Mixed into a small drop of water on the slide

38

If the organisms are growing in a liquid, a small drop

Is spread across the surface of the slide

39

In heat fixation

Developed by Robert Kosh more than 100 years ago, the slide is gently heated by passing the slide, smear up, through the flame of a bunsen burner

40

Chemical fixation involves

Applying a chemical such as methyl alcohol to the smear for one minute

41

Desiccation (drying) and fixation does what to the microorganism?

Kill the microorganism, attaches them firmly to the slide, & generally preserve their shape & size

42

Specimens for scanning electron microscopy are?

Coated not stained

43

Dyes used for microbiological stains for light microscopy are usually?

Salts

44

Salt (usual dye) is composed of?

A positively charged cation and a negatively charged anion. At least one of the two ions in the molecular makeup of dyes is colored; this colored portion of a dye is known as the chromophore

45

Chromophores bind to chemicals via?

Covalent, ionic, or hydrogen bonds. For example methylene blue chloride is composed of a cationic chromophore, methylene blue, and a chloride anion. Because methylene blue is positively charged, it ionically bond to negatively charged molecules in cells, including DNA & many proteins

46

Anionic dyes, for example eosin, bind to?

Positively charged molecules such as amino acids

47

Anionic chromophores are also called?

Acidic dyes because they stain alkaline structures & work best in acidic (low pH) environments.

48

Basic dyes are

Positively charged, cationic chromophores. They combine with & stain acidic structures; further, they work best under basic (higher pH) conditions.

49

In microbiology which dyes are more commonly used more, basic or acid?

Basic dyes because most cells are negatively charged

50

Acidic dyes are used in?

Negative staining

51

Selectively stains membranes because it is lipid soluble and accumulates in phospholipid bilayers

Sudan black

52

Simple stains are?

Composed of a single basic dye, such as crystal violet, safranin, or methylene blue. They are "simple" because they involve no more than soaking the smear in the dye for 30-60 seconds and then rinsing off the slide with water. After carefully blotting the slide you can observe the smear under the microscope

53

Simple stains are used to?

Determine size, shape, and arrangement of cells

54

Differential stains (most common) use?

More than one dye so that different cells, chemicals, or structures can be distinguished when microscopically examined

55

Common differential stains are?

Gram stain, the acid-fast stain, the endospore stain, Gomori methenamine silver stain, and hematoxylin and eosin stain

56

Who developed the Gram stain?

In 1884, the Danish scientist Hans Christian Gram

57

The Gram stain differentiates between?

Two large groups of microorganisms: purple-staining Gram-positive cells and pink-staining Gram-negative cells. These cells differ significantly in the chemical and physical structures of their cell walls

58

Typically, a ______ stain is the first step a medical laboratory technologist performs to identify bacterial pathogens

Gram

59

4 Steps of Gram staining procedure

1) Flood smear with basic crystal violet for 1 min. then rinse w/water. This dye colors all cells
2) Flood smear with iodine solution for 1 min. then rinse w/water. After this step all cells remain purple
3) Rinse smear w/solution of ethanol & acetone (10-30 sec) then rinse w/water. This breaks down thin cell wall of Gram-negative cells allowing stain and mordant to be washed away. These cells are now colorless. Gram-positive cells w/their thick cell walls remain purple
4) Flood smear w/safranin for 1 min. Rinse. This red counterstain provides contrasting color to the primary stain. After this step Gram-negative cells are pink
After final step the slide is blotted dry

60

The primary stain of Gram stain

Crystal violet

61

Iodine is a?

Mordant, a substance that binds to a dye & makes it less soluble

62

Acts as a decolorizing agen

Solution of ethanol and acetone

63

The Gram procedure works best with?

Young cells. Older Gram-positive cells bleach more easily than younger cells & can therefore stain pink, which makes them appear to be Gram-negative cells. Therefore, smears for Gram staining should come from freshly grown bacteria

64

The acid-fast stain is

Another differential stain because it stains cells of the genera Mycobacterium and Nocardia which cause many human diseases, including TB, leprosy, & other lung & skin infections. Cells of these bacteria have large amounts of waxy lipid in their cell walls, so they do not readily stain with the Gram stain

65

Acid-fast stain developed by?

Franz Ziehl and Friedrich Neelsen in 1883

66

Procedure of acid-fast stain

1) Cover smear with piece of tissue paper to retain dye
2) Flood slide w/red primary stain, carbolfuchsin, for several min. while warming over steaming water. This is used to drive the stain through the waxy wall and into the cell
3) Remove paper, cool slide, decolorize smear by rinsing w/solution of hydrochloric acid & alcohol. The bleaching action of acid-alcohol removes color from both non-acid fast cells and the background. Acid-fast cells retain their color because the acid cannot penetrate the waxy wall
4) Counterstain w/ methylene blue, which stains only bleached, non acid-fast cells

67

The Ziehl-Neelsen acid-fast staining procedure results in?

Pink acid-fast cells, which can be differentiated from blue non-acid fast cells, including human cells and tissue

68

The presence of acid-fast bacilli (AFBs) in sputum is indicative of?

Mycobacterial infection

69

Some bacteria-notably those of the genera Bacillus and Clostridium which contain species that cause such disease as anthrax, gangrene, and tetanus- produce?

Endospores

70

Endospores are

Dormant, highly resistant cells form inside the cytoplasm of the bacteria & can survive environmental extremes such as desiccation, heat, & harmful chemicals

71

Endospores cannot be stained by normal staining procedures because?

Their walls are practically impermeable to all chemicals. The Schaeffer-Fulton endospore stain uses heat to drive the primary stain, malachite green, into the endospore. After cooling the slide is decolorized w/water & counterstained with safranin. This staining procedure results in green-stained endospores & red colored vegetative cells

72

Laboratory technicians use two popular stains to stain histological specimens (tissue samples)

Gomori methenamine silver (GMS) stain is commonly used to screen for the presence of fungi & the locations of carbohydrates in tissues. Hematoxylin & eosin (HE) stain, which involves applying the basic dye hematoxylin & the acidic dye eosin, is used to delineate many features of histological specimens, such as the presence of cancer cells

73

Special stains are?

Simple stains designed to reveal special microbial structures

74

There are three types of special stains

Negative stains, flagellar stains, and fluorescent stains

75

Describe the negative (capsule) stain

Most dyes used to stain bacterial cells, such as crystal violet, methylene blue, malachite green, & safranin, are basic dyes. These dyes stain cells by attaching to negatively charged molecules within them

76

What is repulsed by the negative charges on the surface of cells & therefore do not stain them?

Acidic dyes. Such stains are called negative stains because they stain the background & leave cells colorless. Eosin & nigrosin are examples of acidic dyes used for negative staining. A counterstain may be added to color the cells

77

Negative (capsule) stains are used primarily to?

Reveal the presence of negatively charged bacterial capsules. Therefore, they are also called capsule stains. Encapsulated cells appear to have a holo surrounding them

78

Pararosaniline and carbolfuchsin, & mordants, such as tannic acid & potassium alum are applied in a series of steps and are examples of?

Flagellar stains

79

Pararosaniline and carbolfuchsin, & mordants, such as tannic acid & potassium alum are applied in a series of steps and are examples of flagellar stains, these molecules bind to flagella and?

Increase their diameter, & change their color, all of which increase contrast & make them visible

80

Laboratory technicians increase contrast & resolution for transmission electron microscopy by?

Using stains, however these stains are not colored dyes but are chemicals containing atoms of heavy metals such as lead, osmium, tungsten, & uranium, which absorb electrons

81

Electron-dense stains may

Bind to molecules w/in specimens, or they may stain the background. The latter type of negative staining is used to provide contrast for small specimens (viruses & molecules)

82

Stains for electron microscopy can be general, in that they?

Stain most objects to some degree, or they may be highly specific. For example, osmium tetra oxide (OsO4) has an affinity for lipids & is thus used to enhance the contrast of membranes

83

Electron-dense stains can also be linked to?

Antibodies to provide an even greater degree of staining specificity because antibodies bind only to their specific target molecules

84

Reveals size, morphology, & arrangement of cell

Simple stains

85

Uniform purple stain

Crystal violet (simple stain)

86

Uniform blue stain

Methylene blue (simple stain)

87

Differentiates Gram-postivie and Gram-negative bacteria, which is typically the first step in their identification

Gram stain

88

Differential stains. Use two or more dyes to differentiate between cells or structures

1) Gram stain
2) Ziehl-Neelsen acid-fast stain
3) Schaeffer-Fulton endospore stain

89

Distinguishes the genera Mycobacterium and Nocardia from other bacteria

Ziehl-Neelsen acid-fast stain

90

Highlights the presence of endospores produced by species in the genera Bacillus and Clostridium

Schaeffer-Fulton endospore stain

91

Reveals bacterial capsules

Negative stain for capsules

92

Allows determination of number & location of bacterial flagella

Flagellar stain