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[MT 6317] Clinical Chemistry Laboratory > [1S] UNIT 4 Analytical Techniques & Automation > Flashcards

[1S] UNIT 4 Analytical Techniques & Automation Flashcards

1
Q

Four Major Disciplines of Analytic Techniques

A
  1. Spectrometry
  2. Luminescence
  3. Electroanalytic Methods
  4. Chromatography
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2
Q

Is the distance between 2 successive peaks and it is expressed in terms of nanometer (nm)

A

Wavelength

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3
Q

<400 nm =
400-700 nm =
>700 nm =

A

UV
Visible
Infrared

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4
Q

The longer the wavelength, the ___________ the energy

A

lower

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5
Q

The higher the frequency, the _______ the energy

A

higher

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6
Q

Planck’s formula

A

E = hv

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7
Q

BEER’S LAW

Concentration of the unknown substance is ________ ________ to the absorbed light (absorbance or optical density) and ________ ________ to the amount of transmitted light.

A

directly proportional
inversely proportional

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8
Q

T/F: A solution transmits light corresponding in wavelength to its color, and usually absorbs light of wavelengths complementary to its color

A

T

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9
Q

Which wavelength would be absorbed strongly by a yellow-colored solution?
a. 450 nm c. 600 nm
b. 585 nm d. 650 nm

A

A

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10
Q

A blue-colored solution would show highest transmittance at:

a. 475 nm c. 585 nm
b. 525 nm d. 620 nm

A

A

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11
Q

Parts of the Spectrophotometer

A

LEME CPR kya mn yn

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12
Q

Parts of the Spectrophotometer

A

LEME CPR kya mn yn

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13
Q

LIGHT SOURCE: CONTINUUM SOURCE

Tungsten

A

Infrared & Visible Light

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14
Q

LIGHT SOURCE: CONTINUUM SOURCE

Deuterium & Xenon

A

UV

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15
Q

LIGHT SOURCE: LINE SOURCE

Mercury-vapor lamps

A

Visible & UV

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16
Q

Which of the following light source provides wavelengths of light that fall under the infrared region?

a. Mercury
b. Xenon
c. Deuterium
d. Silicon Carbide

A

D

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17
Q

Which of the following light source provides wavelengths of light that fall under the infrared region?

a. Mercury
b. Xenon
c. Deuterium
d. Silicon Carbide

A

D

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18
Q

PARTS OF SPECTROPHOTOMETER

Minimizes unwanted stray of light
Prevents entrance of scattered light on the monochromator system

A

Entrance Slit

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19
Q

ENTRANCE SLIT

  • Any wavelength outside the band transmitted by the monochromator
  • Causes absorbance error
A

Stray Light

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20
Q

ENTRANCE SLIT

  • Limits the maximum absorbance that a spectrophotometer can achieve
  • Most common cause of lost linearity at high analyte concentration
A

Stray Light

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21
Q

PARTS OF SPECTROPHOTOMETER

It isolates specific or individual wavelength of light

A

Monochromator

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22
Q

PARTS OF SPECTROPHOTOMETER

Kinds of Monochromator

A
  1. Prisms
  2. Diffraction Gratings
  3. Filters
  4. Holographic Gratings
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23
Q

PARTS OF SPECTROPHOTOMETER

Controls the width of the light beam (bandpass)

A

Exit Slit

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24
Q

PARTS OF SPECTROPHOTOMETER

Furthermore, a bandpass can be described as the ________________.

A

Exit Slit; total range of wavelengths transmitted

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25
Q

PARTS OF SPECTROPHOTOMETER

Allows only a narrow fraction of the spectrum to reach the sample cuvette

A

Exit Slit

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26
Q

PARTS OF SPECTROPHOTOMETER

holds the solution whose concentration is to be measured

A

Cuvette

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27
Q

PARTS OF SPECTROPHOTOMETER

Kinds of Cuvette

A
  1. Alumina Silica Glass
  2. Quartz Plastic
  3. Borosilicate glass
  4. Soft glass
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28
Q

PARTS OF SPECTROPHOTOMETER: KINDS OF CUVETTE

Capable of withstanding light with wavelength of 350-2000 nm

A

Alumina Silica Glass

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29
Q

PARTS OF SPECTROPHOTOMETER: KINDS OF CUVETTE

Visible-UV

A

Quartz Plastic

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30
Q

PARTS OF SPECTROPHOTOMETER

detects and converts transmitted light into photoelectric energy

A

Photodetector

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31
Q

PARTS OF SPECTROPHOTOMETER

Kinds of Photodetectors

A
  1. Photocell
  2. Phototube
  3. Photomultiplier tube
  4. Photodiode
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32
Q

PARTS OF SPECTROPHOTOMETER: PHOTODETECTORS

Most sensitive; used in fluorometric assay

A

Photomultiplier Tube

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33
Q

Compare single beam and double beam

A

Single Beam = 1 cuvet
Double Beam = 2 cuvets (Sample & Reference)

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34
Q

DOUBLE BEAM

2 photodetectors & cuvettes

A

In Space

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35
Q

DOUBLE BEAM

Only 1 photodetector

A

In Time

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36
Q

INSTRUMENTATION

Measures light emitted by one atom burned in a flame

A

Flame Emission Photometry

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37
Q

INSTRUMENTATION PRINCIPLE

Measures the excitation of electrons from lower to higher state

A

Flame Emission Photometry

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38
Q

INSTRUMENTATION PHOTODETECTOR & LIGHT SOURCE

Photo cell
Flame

A

Flame Emission Photometry

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39
Q

INSTRUMENTATION PHOTODETECTOR & LIGHT SOURCE

Photo cell
Flame

A

Flame Emission Photometry

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40
Q

INSTRUMENTATION

Measures light absorbed by one atom in a dissociated by heat

A

Atomic Absorption Spectrophotometry

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41
Q

INSTRUMENTATION

Measures light absorbed by one atom in a dissociated by heat

A

Atomic Absorption Spectrophotometry

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42
Q

INSTRUMENTATION

Most sensitive & specific for electrolytes and trace elements

A

Atomic Absorption Spectrophotometry

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43
Q

INSTRUMENTATION PRINCIPLE

Dissociation of subatomic bonds in electrolytes

A

Atomic Absorption Spectrophotometry

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44
Q

INSTRUMENTATION PHOTODETECTOR & LIGHT SOURCE

Photomultiplier Tube
Hollow Cathode Lamp

A

Atomic Absorption Spectrophotometry

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45
Q

When measuring calcium by atomic absorption spectrophotometry, which is required?

a. An organic extraction reagent to deconjugate calcium from protein
b. An internal standard
c. A magnesium chelator
d. Lanthanum oxide to chelate phosphates

A

D

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46
Q

INSTRUMENTATION PRINCIPLE

The unknown sample is made to react with a known solution in the presence of an indicator

A

Volumetric (Titremetric)

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47
Q

INSTRUMENTATION: VOLUMETRIC

For determination of fluoride concentration in the bloodstream

A

Schales and Schales Method

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48
Q

INSTRUMENTATION: VOLUMETRIC

For determination of fluorife concentration in the bloodstream

A

Schales and Schales Method

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49
Q

INSTRUMENTATION: VOLUMETRIC

Chelates calcium

A

EDTA Titration Method

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50
Q

INSTRUMENTATION PRINCIPLE

  • It determines the amount of scattered light by a particulate matter suspended in a turbid solution
  • Measures protein in a sample
A

Nephelometry

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51
Q

INSTRUMENTATION: NEPHELOMETRY

Light Scattering depends on the following factors?

A

a. Particle Size
b. Wavelength

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52
Q

INSTRUMENTATION

➢ Measures the amount of Antigen-Antibody Complexes.
➢ Measures the angle at 15-90 degrees.

A

Nephelometry

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53
Q

Used to descrive a molecule that invoke an immune response

A

Antigens

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54
Q

Most anti antigens to least (d k alam bt gnyn pero yn nsa notes k bhala kau)

A
  1. Proteins
  2. Polysaccharides
  3. Nucleic Acids & Lipids
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55
Q
  1. Direct neutralization
  2. Opsonization
  3. Complement activation
  4. Somatization
  5. Control of inflammatory response
A

NEPHELOMETRY: Antibody Molecule

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56
Q

INSTRUMENTATION PRINCIPLE

Measures reduction (not specific protein) in light transmission by one particle formation

A

Turbidimetry

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57
Q

This is the process of separating the charged constituents of a sample by means of an electrical current

A

Electrophoresis

58
Q

ELECTROPHORESIS

Migration of charged macromolecules in the presence of an electrical power through a porous support:
a. Paper
b. Cellulose acetate
c. Agarose gel
d. Polysaccharide

A

Zone Electrophoresis

59
Q

ELECTROPHORESIS

Has a net charge that can be either positive or negative depending on pH conditions

A

Amphoteric

60
Q

ELECTROPHORESIS

T/F: Amphoteric Electrophoresis is always done in acidic solution (2.0) to become negatively charged

A

F; alkaline solution (9.0)

61
Q

ELECTROPHORESIS

Movement of buffer ions & solvent relatives to the fixed support

A

Electroendosmosis/Endosmosis

62
Q

ELECTROPHORESIS

Migration of small charged ions

A

Iontophoresis

63
Q

5 Components of Electrophoresis

A
  1. The driving force
  2. The support medium
  3. The buffer
  4. The sample
  5. The detecting system
64
Q

3 Electrophoresis Support Media

A
  1. Cellulose acetate
  2. Agarose gel
  3. Polyacrylamide gel
65
Q

6 Electrophoresis Procedures (Proteins)

A
  1. Loading of patient sample
  2. Electrophoretic migration
  3. Wash and Fix
  4. Staining
  5. Visualization (Qualitative Result)
  6. Quantification
66
Q

5 Electrophoresis Fractions of Separated Proteins

A
  1. Albumin
  2. Alpha1
  3. Alpha2
  4. Beta
  5. Gamma globulins
67
Q

7 Electrophoresis Stains for Visualization of Bands

A
  1. Amido Black
  2. Ponceau S
  3. Oil Red O
  4. Sudan Black
  5. Fat Red 7B
  6. Coomassie Blue
  7. Gold/Silver Stain
68
Q

ELECTROPHORESIS STAINS FOR VISUALIZATION OF BANDS

For plasma proteins

A

Amido Black

69
Q

ELECTROPHORESIS STAINS FOR VISUALIZATION OF BANDS

Proteins in very minute amounts

A

Gold/Silver Stain

70
Q

What dye maybe used for staining protein
bands following electrophoresis?

a. Fat red 7B
b. Sudan black B
c. Ponceau S
d. Oil red O

A

c. Ponceau S

71
Q

Which of the following reagents can be used to measure protein present in the cerebrospinal fluid?

a. Biuret
b. Coomassie brilliant blue
c. Ponceau S
d. Bromcresol green

A

b. Coomassie brilliant blue

72
Q

INSTRUMENTATION PRINCIPLE

Involves the separation of soluble materials in a solution by specific chemical and physical differences through:
- Rate of diffusion
- Solubility
- Size
- Ionic charges

A

Chromatography

73
Q

CHROMATOGRAPHY

Diffusion of molecules in 2D-plane system

A

Planar Form Chromatography

74
Q

PLANAR FORM CHROMATOGRAPHY

Fractionation of sugar and amino acids through whattaman paper sorbent

A

Paper Chromatography

75
Q

PLANAR FORM CHROMATOGRAPHY

Used for TDM to separate drug molecules through plastic plates sorbent

A

Thin Layer Chromatography

76
Q

CHROMATOGRAPHY

Diffusion of analytes in 3D-multidirectional system

A

Column Form Chromatography

77
Q

COLUMN FORM CHROMATOGRAPHY

Elution of volatile compounds based on boiling point, used to separate steroids, lipids, alcohols

A

Gas Chromatography

78
Q

COLUMN FORM CHROMATOGRAPHY: GAS

Differences in absorption of gases sat solid phase surfaces

A

Gas Solid Chromatography

79
Q

COLUMN FORM CHROMATOGRAPHY: GAS

Differences in solute partitioning between gaseous mobile phase vs liquid stationary phase

A

Gas Liquid Chromatography

80
Q

COLUMN FORM CHROMATOGRAPHY

Distribution of solute betweem liquid mobile phase vs liquid stationary phase

A

Liquid Chromatography

81
Q

COLUMN FORM CHROMATOGRAPHY: LIQUID

Most widely used, uses pressure for fast separation, fractionation of drugs, hormones & Hgb variants

82
Q

COLUMN FORM CHROMATOGRAPHY: LIQUID

Separate non-volatile substances in human body fluids, complementary method to GC-MS as it is used to confirm positive results from screening illicit drugs

83
Q

Fragmentation and ionization of molecules using suitable source of energy (ex: CT scan)

A

Mass Spectroscopy

84
Q
  • Advantage: Requires smaller sample volume
  • Disadvantage: Destructive
A

Mass Spectroscopy

85
Q

Determine the structure of organic compound (ex: MRI)

A

Nuclear Magnetic Resonance Spectroscopy

86
Q
  • Advantage: Non-destructive
  • Disadvantage: Requires larger sample volume
A

Nuclear Magnetic Resonance Spectroscopy

87
Q

SEPARATION TECHNIQUES

Migration is based on electrical charge

A

Electrophoresis

88
Q

SEPARATION TECHNIQUES

Migration is based on physical/chemical properties

A

Chromatography

89
Q

SEPARATION TECHNIQUES

Migration is through a pH gradient

A

Isoelectric Focusing

90
Q

SEPARATION TECHNIQUES

Migration is through an electro-osmosis flow

A

Capillary Electrophoresis

91
Q

INSTRUMENTATION PRINCIPLE

Amount of light emitted by a molecule after excitation by electromagnetic radiation

A

Fluorometry

92
Q

INSTRUMENTATION

  • Measures amount of light intensity present over a dark background
  • Utilizes 2 monochromators
A

Fluorometry

93
Q

INSTRUMENTATION

Advantage: More specific & 1000x more sensitive than spectro

A

Fluorometry

94
Q

INSTRUMENTATION

Light source of Fluorometry

A

Mercury
Xenon
UV Lights

95
Q

INSTRUMENTATION

Photodetector of Fluorometry

A

PMT / Phototube

96
Q

FLUOROMETRY

T/F: Affected by photobleaching - pH, temp, UV, and chemical changes

A

T; quenching & photobleaching r same bananas

97
Q

FLUOROMETRY

T/F: Quenching is the reduction or limitation of a particle in an excited state

98
Q

↑ Light Exposure = _ Fluoroscence

99
Q

↑ Unwanted light exposure = _ Fluoroscence

100
Q

↑Temperature = _ Fluoroscence

101
Q

↑ Quenching = _ Fluoroscence

102
Q

↑ Absorbing analytes = _ Fluoroscence

103
Q

Detection systems used in real time PCR are based on:

a. radioactivity
b. chemiluminescence
c. fluorescence
d. colorimetry

A

c. fluorescence

104
Q

INSTRUMENTATION PRINCIPLE

Measurement of luminescence produced by chemical reaction producing light emission

A

Chemiluminescence

105
Q

INSTRUMENTATION PRINCIPLE

Measurement of luminescence produced by chemical reaction producing light emission

A

Chemiluminescence

106
Q

INSTRUMENTATION

Measures amount of light emission based on chemical or electrochemical reaction

A

Chemiluminescence

107
Q

INSTRUMENTATION

More sensitive than fluorometry and spectrophotometry

A

Chemiluminescence

108
Q

INSTRUMENTATION

More sensitive than fluorometry and spectrophotometry

A

Chemiluminescence

109
Q

Which of the following components is not needed in a chemiluminescent immunoassay analyzer?

a. Source lamp
b. Monochromator
c. Photodetector
d. Wash station

A

a. Source lamp

110
Q

INSTRUMENTATION PRINCIPLE

● changes in colligative property of solutions that occur due to variations in particle concentrations
● Measurement of the osmolarity of an aqueous solution
○ (Ex.) osmotic particles: Glucose, BUN, Sodium

111
Q

Increased osmolarity = Increased osmotic pressure, boiling point = _______ vapor pressure, freezing point

112
Q

ELECTROCHEMISTRY

● Measurement of electrical potential due to the activity of free ions
● Change of voltage indicates analyte activity (potential means voltage)

A

POTENTIOMETRY

113
Q

ELECTROCHEMISTRY

● Reference electrodes
○ Calomel
○ Silver-Silver Chloride
○ H+ Electrode
● Measures: pH and pCO2
● Relies on the concept of Nerst Equation

A

POTENTIOMETRY

114
Q

ELECTROCHEMISTRY

● Electrochemical transducer capable of responding to one given ion
● Very sensitive and selective, yet non-specific
● A type of potentiometer

A

ION SELECTIVE ELECTRODE

115
Q

ION SELECTIVE ELECTRODE MEMBRANES

For measuring sodium

A

Glass aluminum silicate

116
Q

ION SELECTIVE ELECTRODE MEMBRANES

For potassium measurement

A

Valinomycin gel

117
Q

ION SELECTIVE ELECTRODE MEMBRANES

For calcium and lithium

A

Organic liquid

118
Q

● Direct ISE: uses ______ sample
● Indirect ISE: uses _______ sample

A

● diluted sample
● undiluted sample

118
Q

ELECTROCHEMISTRY

● Measures: chloride test (CSF, serum, sweat)
● Governed by Faraday’s Law
○ For monitoring cystic fibrosis

A

COULOMETRY

119
Q

ELECTROCHEMISTRY

● Measurement of the amount of electricity at fixed potential coulombs
● Electrochemical titration in which the titrant is electrochemically generated and the end point is detected by amperometry

A

COULOMETRY

120
Q

ELECTROCHEMISTRY

Measures: pO2, glucose, chloride, peroxidases

A

AMPEROMETRY

121
Q

ELECTROCHEMISTRY

● Current flow produced by oxidation reaction

A

AMPEROMETRY

122
Q

ELECTROCHEMISTRY

● Measurement of differences in current at a constant voltage (Clarke’s Polarographic Electrode)
● A type of amperometry, uses Ilkovic Equation

A

POLAROGRAPHY

123
Q

ELECTROCHEMISTRY

● Measurement of current after a potential is applied to an electrochemical cell
● Allows sample to be pre-concentrated, thus minimal analyte

A

VOLTAMMETRY

124
Q

ELECTROCHEMISTRY

Anodic stripping voltammetry: specifically utilized for lead and iron studies

A

VOLTAMMETRY

125
Q

AUTOMATION IN THE CLINICAL LABORATORY

● Samples flow through a common vessel or pathway
● Liquids are pumped through a system of continuous tubing

A

CONTINUOUS FLOW ANALYZER

126
Q

AUTOMATION IN THE CLINICAL LABORATORY

Air bubbles serve as separating and cleaning media

A

CONTINUOUS FLOW ANALYZER

127
Q

AUTOMATION IN THE CLINICAL LABORATORY

Allow random access or STAT capabilities

A

DISCRETE ANALYZER

127
Q

AUTOMATION IN THE CLINICAL LABORATORY

● Each sample-reagent mixture is handled in its own reaction vessel
● Employs a variety of syringe pipettes to aspirate and dispense samples (2-6 uL) and reagents

A

DISCRETE ANALYZER

128
Q

AUTOMATION IN THE CLINICAL LABORATORY

● Uses the force generated by centrifugation to transfer specimen and reagents
● Major advantage: Batch analysis

A

CENTRIFUGAL ANALYZER

129
Q

GLASSWARES

  • Up to 515 degrees celcius
  • Most common (heating & sterilization)
A

Borosilicate Glass (Pyrex, Kimax)

130
Q

GLASSWARES

  • High thermal resistance
  • Low alkali content (free from magnesium lime)
A

Borosilicate Glass (Pyrex, Kimax)

131
Q

GLASSWARES

  • Less thermal resistance than borosilicate
  • High alkali resistance
A

Boron-free Glass (Soft Glass)

132
Q

GLASSWARES

  • 6x stronger than borosilicate
  • Strengthened chemically than thermally
A

Corex (Corning, Alumina-silicate Glass)

133
Q

GLASSWARES

  • Up to 900C
  • High thermal-drastic heat shock resistance
  • Extremely chemical (acid, alkali) resistant
A

Vycor (Corning)

134
Q

GLASSWARES

  • Poor thermal resistance
  • Disposable
A

Flint Glass (Soda-lime Glass)

135
Q

GLASS PIPET ACCDNG TO CALIB DESIGN

Holds volume but don’t dispense exact amount

136
Q

GLASS PIPET ACCDNG TO CALIB DESIGN

Delivers exact amount

137
Q

GLASS PIPET ACCDNG TO DRAINAGE CHARAC

Etched ring, need to blow last drop to get exact volume

138
Q

GLASS PIPET ACCDNG TO DRAINAGE CHARAC

Etched ring, gravity auto-drains the liquid

A

Self-draining

139
Q

NEPHELOMETRY

If a particular analyte has a larger size than the wavelength of light, it will scatter the light in a forward direction in an angle of 90 degrees or lower

A

Mie scatter

140
Q

INSTRUMENTATION

Light source of Nephelometry

[MT 6317] Clinical Chemistry Laboratory (10 decks)

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