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Clinical Lab Science Lab > Chapter 7 > Flashcards

Chapter 7 Flashcards

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1
Q

General methods for common automated and manual assays include the use of- (6)

A

-photometry
-spectrometry
-ion-selective electrodes (ISEs)
-electrophoresis
-nephelometry
-immunoassays

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2
Q

measurement techniques fall into one of four categories-

A

-Spectrometry
-Luminescence
-Electroanalytical methods
-Chromatography

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3
Q

Photometry employs ____ & ____ ______ to determine the concentrations of various substances.

A

-color
-color variation

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4
Q

the measurement of the luminous intensity of light, or the amount of luminous light falling on a surface from a light source-

A

photometry

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5
Q

The concentration of an unknown sample is determined by measuring-

A

light absorption at a certain wavelength & comparing it with light absorption by standard solutions measured at the same time & wavelength

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6
Q

Light is a type of-

A

radiant energy

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7
Q

light travels in the form of-

A

waves

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8
Q

The wavelength of light is the distance between-

A

waves

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9
Q

the term light is used to describe-

A

radiant energy with wavelengths visible to the human eye or bordering on those visible to the human eye

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10
Q

Electromagnetic radiation includes a spectrum of energy on the left side- (3)

A

-short-wavelength
-highly energetic gamma rays
-x-rays

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11
Q

Electromagnetic radiation includes a spectrum of energy on the right side-

A

wavelengths of radio frequencies

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12
Q

Visible light passes between these frequencies, with the color violet at about ____ wavelength & red at ___ wavelength-

A

-400-nm
-700-nm

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13
Q

appx. wavelength of not visible (ultraviolet light)-

A

<380 nm

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14
Q

appx. wavelength of violet light-

A

380 - 440 nm

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15
Q

appx. wavelength of blue light-

A

440 - 500 nm

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16
Q

appx. wavelength of green light-

A

500 - 580 nm

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17
Q

appx. wavelength of yellow light-

A

580 - 600 nm

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18
Q

appx. wavelength of orange light-

A

600 - 620 nm

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19
Q

appx. wavelength of red light-

A

620 - 750

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20
Q

appx. wavelength of not visible (infrared light)-

A

> 750

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21
Q

Beer’s law states that the concentration of a substance is directly proportional to-

A

the amount of light absorbed

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22
Q

Beer’s law states that the concentration of a substance is inversely proportional to-

A

the logarithm of the transmitted light

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23
Q

Beer’s law is the basis for the use of photometry in-

A

quantitative measurement

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24
Q

In absorbance spectrophotometry, the absorbance units or values for several different concentrations of a standard solution are determined by-

A

spectrophotometry & plotted on graph paper

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25
Q

The resulting graph in absorbance spectrophotometry is known as- (3)

A

-standard calibration curve
-standard curve
-Beer-Lambert (Beer’s) law plot

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26
Q

Absorbance is an expression of-

A

amount of light absorbed by a solution

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27
Q

absorbance =

A

2 minus the logarithm of the percent transmittance

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28
Q

units used to express the readings obtained by the electronic measuring device are either-

A

-absorbance units
-percent transmittance units

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29
Q

Absorbance values are directly proportional to the concentration of-

A

the solution & can be plotted on graph paper to give a straight line

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30
Q

Percent transmittance is the amount of light that passes through a-

A

colored solution compared with the amount of light that passes through a blank or standard solution

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31
Q

Transmitted light does not decrease in direct proportion to-

A

concentration or color intensity of the solution being measured

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32
Q

Percent transmittance readings plotted against concentration will not give-

A

a straight line on linear graph paper, but will on semilogarithmic graph paper

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33
Q

Standard curves are defined as-

A

a graph with absorption (A) or %T plotted on the y-axis (vertical axis), & increasing concentrations of standard along the x-axis (horizontal axis)

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34
Q

If Beer’s law is followed, the resulting line representing absorbance versus concentration will be-

A

straight

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35
Q

Once a standard curve is developed for a test method on a particular spectrophotometer, it should be checked-

A

periodically to determine that it is still good

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36
Q

Semilogarithmic (semilog) graph paper is used to plot-

A

%T readings from the photometer

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37
Q

The horizontal axis (x-axis) of semilog graph paper is a ______ scale-

A

linear

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38
Q

The vertical axis (y-axis) of semilog graph paper is a _______ scale-

A

log

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39
Q

Concentrations of the standard solutions are plotted on the ______ axis-

A

horizontal

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40
Q

The transmittance or absorbance readings from the photometer are plotted along the ______ axis-

A

vertical

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41
Q

Criteria for a good standard curve- (3)

A

-The line is straight
-The line connects all points
-The line goes through the origin, or intersect, of the two axes

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42
Q

plotting a standard curve- (3)

A

-Note the intervals on the graph paper
-Axes must be properly labeled
-Review sources of error

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43
Q

Once the standard curve has been plotted, it is used to calculate-

A

concentrations of any unknowns that were included in the same batch as the standards used to make the graph

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44
Q

Parts essential to all spectrophotometers- (4)

A

-Light source
-Wavelength isolator (filter)
-Cuvettes (absorption cells or photometer tubes)
-Electronic measuring device (commonly a photoelectric cell plus galvanometer)

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45
Q

Quality control tests for spectrophotometers- (3)

A

-Wavelength accuracy
-Stray light
-Linearity

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46
Q

A beam of light is directed at a flat surface, and the amount of light reflected is measured in a-

A

reflectance spectrophotometer

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47
Q

Light reflected from the surface of a colorimetric reaction is used to measure the amount of-

A

unknown colored product generated in the reaction

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48
Q

Quality control for single-test instruments is integrated into the instruments by-

A

manufacturer

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49
Q

Manufacturing processes, shipping & handling, or storage problems can affect-

A

measurements

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50
Q

A lamp generates light, which passes through- (2)

A

-filter
-series of slits

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51
Q

A lamp generates light, which is focused on-

A

the test surface

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52
Q

Some light is absorbed by-

A

the filter

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53
Q

the remaining light that isn’t absorbed is-

A

reflected

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54
Q

The reflected light then passes through a series of-

A

slits & lenses & on to the photodetector device, where the amount of light is measured & recorded as a signal

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55
Q

Common point-of-care testing and self-testing instruments include those for-

A

quantitation of blood glucose (employing single-test methodology) in maintaining good diabetic control

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56
Q

In urinalysis testing, various instruments use-

A

dry-reagent reflectance spectrophotometry

57
Q

Chemistry and therapeutic drug-monitoring analyzer systems also employ-

A

single-test methodology technology

58
Q

Fluorescent light is the result of-

A

the absorbance of a photon of radiant energy by a molecule

59
Q

When excess energy is ejected as a photon, the result is-

A

fluorescence emission, which can be measured

60
Q

The intensity of the fluorescence is determined by using a-

A

fluorometer, sometimes called a spectrofluorometer or fluorescence spectrophotometer

61
Q

Only a few compounds can fluoresce, and of those that do, not all photons absorbed will be converted to-

A

fluorescent light

62
Q

Nephelometry is the measurement of-

A

light that has been scattered

63
Q

Tubidimetry is the measurement of-

A

a loss in the intensity of light transmitted through a solution because of the light being scattered. (The solution becomes turbid)

64
Q

Turbidimetry will measure light that is-

A

scattered, not absorbed or reflected by the particles in the suspension

65
Q

Nephelometers are used to detect-

A

the amount of light scattered

66
Q

When specific antigen-coated latex particles acting as reaction intensifiers are agglutinated by their-

A

corresponding antibody

67
Q

the increased light scatter of a solution can be measured by-

A

nephelometry as the macromolecular complexes form

68
Q

Formation of a macromolecular complex is a prerequisite for-

A

nephelometric protein quantitation

69
Q

The amount of scattered light is proportional to-

A

the number of insoluble complexes

70
Q

the amount of scattered light can be quantitated by comparing-

A

unknown patient values with standards of known protein concentration

71
Q

An infrared high-performance light-emitting diode is used as-

A

the light source

72
Q

advantages of Nephelometry- (2)

A

-automated system that is rapid, reproducible, relatively simple to operate, & very common in higher-volume labs.
-has many applications in immunology laboratory

73
Q

disadvantages of Nephelometry- (2)

A

-high initial equipment cost
-Interfering substances may cause protein denaturation & erroneous test results

74
Q

Laser light is the most common light source used in-

A

flow cytometers

75
Q

laser light is the most common light source used in flow cytometers because of its properties of-

A

-intensity
-stability
-monochromaticity

76
Q

Cells in suspension are stained with-

A

an appropriate fluorochrome, such as an immunologic reagent or a dye that stains a specific component

77
Q

Fluorescent dyes used in flow cytometry bind or react with the-

A

cellular component of interest

78
Q

The stained cells then pass through the-

A

laser beam in single file

79
Q

The stained cells then pass through the laser beam in single file. The laser activates- (2)

A

-dye
-cell fluoresces

80
Q

Applications of labels include- (3)

A

-enzyme immunoassays (EIAs)
-chemiluminescence
-fluorescent substances

81
Q

Chemiluminescent reactions have replaced-

A

most radioimmunoassays in the clinical lab

82
Q

Immunoassays use-

A

antigen-antibody reactions

83
Q

An antigen may be used as a reagent to detect-

A

the presence of antibodies in the serum of a patient

84
Q

Immunoassays can be divided into- (2)

A

-heterogeneous immunoassays
-homogeneous immunoassays

85
Q

Homogeneous immunoassays are faster and easier to-

A

automate than heterogeneous immunoassays & have competitive formats

86
Q

Enzyme immunoassay (EIA) can test for- (2)

A

-specific antigen
-antigen-specific antibody

87
Q

Enzyme-linked immunosorbent assay (ELISA), is designed to detect-

A

antigens or antibodies by producing an enzyme-triggered color change

88
Q

EIAs for ____ & _____ detection-

A

-antigen detection
-antibody detection

89
Q

Lateral or vertical-flow immunoassays also known as-

A

immunochromatography assay

90
Q

immunochromatography is a versatile and rapid method for-

A

visual detection of antigen in a blood sample on a test strip

91
Q

Lateral or vertical-flow immunoassays has components of-

A

a test strip

92
Q

Immunofluorescent technique labeling can demonstrate-

A

the complexing of antigens & antibodies

93
Q

in Direct immunofluorescent assay
A conjugated antibody is used to detect-

A

antigen-antibody reactions at a microscopic level

94
Q

Inhibition immunofluorescent assay is a blocking test in which-

A

an antigen is first exposed to unlabeled antibody, then to labeled antibody, & is finally washed and examined

95
Q

Indirect immunofluorescent assay uses-

A

immunoglobulins’ reaction with antiimmunoglobulins

96
Q

Immunofluorescent techniques- (3)

A

-Time-resolved fluoroimmunoassay
-Fluorescence polarization immunoassay
-Fluorescence in situ hybridization

97
Q

alternative Immunofluorescent Labeling Techniques- (2)

A

-Signal amplification technology
-Magnetic labeling technology

98
Q

Chemiluminescence is the emission of light by -

A

molecules in an excited state with a limited amount of emitted heat (luminescence) as the result of a chemical reaction

99
Q

New chemiluminescence systems use-

A

chemiluminescent labels & substrates rather than older fluorescent labels and detection systems

100
Q

New chemiluminescence systems are used to detect- (4)

A

-proteins
-viruses
-oligonucleotides
-genomic nucleic acid sequences

101
Q

proteins, viruses, oligonucleotides, and genomic nucleic acid sequences in- (4)

A

-immunoassays
-DNA probe assays
-DNA sequencing
-electrophoresis

102
Q

Potential Benefits of Immunoassay Automation have the ability to-

A

provide better service with less staff

103
Q

Potential Benefits of Immunoassay Automation save on- (4)

A

-controls
-duplicates
-dilutions
-repeats

104
Q

Potential Benefits of Immunoassay Automation eliminates- (2)

A

-radioactive labels
-associated regulations

105
Q

Immunoassay Automation has a better- (2)

A

-shelf life of reagents, with less disposal caused by outdating
-sample identification with bar code labels & primary tube sampling

106
Q

Immunoassay Automation of sample delivery is-

A

possible

107
Q

Amplification techniques in molecular biology
Polymerase chain reaction (PCR) is a molecular diagnostic technique that-

A

amplifies low levels of specific DNA sequences in a sample to higher quantities suitable for further analysis

108
Q

Other amplification techniques- (4)

A

-Strand displacement amplification
-Transcription-mediated amplification
-Nucleic acid sequence–based amplification
-Ligase chain reaction nucleic acid amplification

109
Q

Analysis of amplification products- (2)

A

-Conventional analysis & other techniques
-DNA sequencing & hybridization techniques

110
Q

Blotting protocols-

A

Western blot

111
Q

Microarrays are miniature gene fragments attached to-

A

glass chips

112
Q

microarrays product of bonding or direct synthesis of-

A

numerous specific DNA probes on a stationary, often silicon-based support

113
Q

microarrays glass chip may be tailored to-

A

particular disease processes

114
Q

the term electrochemistry is used when-

A

chemical energy is converted to an electrical current (a flow of electrons) in a galvanic cell

115
Q

Electrochemical reactions are characterized by-

A

a loss of electrons (oxidation) at the positive pole (anode) & a simultaneous gain of electrons (reduction) at the negative pole (cathode).

116
Q

Electroanalytical chemistry uses electrochemistry for-

A

analysis purposes, such as to measure ions, drugs, hormones, metals, and gases

117
Q

Potentiometry compares-

A

the potential of one electrode to another

118
Q

Ion-selective electrodes (ISEs) are designed to be-

A

sensitive toward individual ions

119
Q

The relationship between the ionic concentration (activity) and the electrode potential is given by-

A

the Nernst equation

120
Q

The term pH refers to the concentration of-

A

hydrogen ions ([H+], also called protons) in a solution

121
Q

For aqueous solutions, the scale ranges from-

A

0 to 14, with pure water in the middle at 7

122
Q

The more acid a solution-

A

the lower is the pH (0 to 6.9)

123
Q

alkaline solutions come in at-

A

the high end of the scale 7.1 to 14

124
Q

Types of meters- (3)

A

-Analog meters
-Digital meters
-Self-calibrating, direct-reading ion meters

125
Q

Coulometry measures-

A

the amount of current passing between two electrodes in an electrochemical cell

126
Q

The principle of coulometry involves the application of-

A

a constant current to generate a titrating agent

127
Q

the time required to titrate a sample at constant current is measured and-

A

is related to the amount of analyte in the sample

128
Q

The amount of current is directly proportional to the amount of-

A

substance produced or consumed by the electrode

129
Q

Electrophoresis is the migration of-

A

charged solutes or particles in an electrical field

130
Q

When charged particles are made to move-

A

differences in molecular structure can be seen because different molecules have different velocities in an electrical field

131
Q

The assay using electrophoresis involves-

A

the movement of charged particles when an external electrical current is produced in a liquid environment

132
Q

Electrophoresis is a technique for-

A

separation and purification of ions, proteins, and other molecules of biochemical interest

133
Q

In chromatography, mixtures of solutes dissolved in a common solvent are separated from one another by-

A

a differential distribution of the solutes between two phases

134
Q

two main categories of chromatography are-

A

-gas chromatography
-liquid chromatography

135
Q

chromatography-

A

in which the solute phase is in a gaseous state

136
Q

liquid chromatography-

A

in which the solute phase is a solution or liquid

137
Q

The same steps needed to perform an analysis from the central laboratory are needed for- point-of-care testing (POCT)

A

point-of-care testing (POCT)

138
Q

steps needed to perform analysis from the central laboratory- (5)

A

-Instrument validation
-Periodic assay calibration
-Quality control testing
-Operator training
-Proficiency testing

Clinical Lab Science Lab (4 decks)

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