INTROS Flashcards
________Process in which ______ force is used to separate solid matter from a liquid suspension
- [#] mins centrifugation of blood
Centrifugation
centrifugal
1O mins
Separation of solids from liquids. However, ________ is only occasionally used in today’s laboratory
Filtration
paper filtration
Separating macromolecules from a solvent or smaller substance. It became popular when used in conjunction with the _________ system in the 1970s.
Dialysis
Technicon Autoanalyzer
Rule in Gaussian Distribution curve
68-95-99 RULE
Summarizes the relationships between the area under the Gaussian distribution curve and the SD.
68-95-99 rule
The reference range is usually set at [#] % limit
95%
Given any data in the Gaussian curve:
➢ _____% of the data will fall between the +/-1SD from the mean.
➢ _____% of the data will fall between the +/- 2SD from the mean.
➢ _____% will fall between +/- 3SD from the mean.
68.3%
95.4%
99.7%
FOR +1SD/2SD/3SD: MEAN ________
FOR -1SD/2SD/3SD: MEAN ________
+(SD) (1)/(2)/(3)
– (SD) (1)/(2)/(3)
A common method to assess the determination of control materials over time is by the use of__________/________
Levey-Jennings chart
Shewhart plot
___________it is the most common method and its generally accepted minimum protocol is: [Frequency]
Levey-Jennings control chart
Shewhart plot
2O times on 2O different days
Westgard rules in random error
12S warning rule
13S reject
R4S
Westgard rules in systematic error
22s
41s
1OX
One control observation exceeding the mean +/-2s. A warning rule that
initiates testing of control data by other rules
12s
One control observation exceeding the mean +/-3s.
13S
Allows high sensitivity to
random error
13S
Two control observations consecutively exceeding the same +2s or -2s. Allows high sensitivity to systematic error
22s
One control exceeding the +2s and another exceeding the -2s. Allows
detection of random error
R4s
Four control observations consecutively exceeding the same +1s or -1s. Allows detection of systematic error
41s
Ten consecutive control observations falling on one side or the other of the
mean (no requirement for SD size). Systematic error
10x
___________
Methodology is the combination of ________quality management, developed by Motorola, with _____ manufacturing strategy, pioneered by ______, to provide tangible metrics for quality improvement. In its simplest form.
Lean Six Sigma
Six sigma
Lean
Toyota
asks the questions, how can this process be improved?
Six sigma
asks the question, does this process or steps need to exist?
Lean
Together, as Lean Six Sigma, they are being increasingly used to reduce error ________and waste _______within the health-care system
Six Sigma
Lean
__________methodology is the quality improvement team’s project management road map.
DMAIC [Define, Measure, Analyze, Improve, Control]
Problems & Objectives
Define
What do we need to improve?
Measure
Process and Factors of influence
Analyze
Implement improvement
Approve
Assure that improvement will sustain
Control
In Beer’s Law, the concentration of the unknown substance is directly proportional to ________ and inversely proportional to _______.
absorbance
transmittance
What law is this?
Under what principles of Instrumentation?
Sample = Absorbance = %T
Beer’s Law
Spectrophotometry
What are the Photometric Methods
Spectrophotometry
AAS
Fluorometry
Chemiluminescence
Turbidimetry
Nephelometry
Chemical reaction produces colored substance that absorbs light of a specific wavelength. Amount of light absorbed is directly proportional to concentration of analyte
Spectrophotometry
COMPONENT PARTS of Spectrophotometry:
Light sources: [2]
Monochromators: [1]
Tungsten lamp [visible range]
Deuterium lamp [UV]
Diffraction grafting
COMPONENT PARTS of Spectrophotometry aside from light sources and monochromators
Cuvette
Photodetector
Readout device
Hollow cathode lamp with cathode made of analyte produces wavelength specific for analyte.
AAS
A photometric that is sensitive and is used for trace metals
AAS
Measures light absorbed by ground-state atoms.
AAS
Component parts:
Atomizer
Flame
Monochromator
Hallow cathode lamp
Mixing chamber
Chopper
Detector
Readout device
AAS
Detector at 90* to light source so that only emitted light by the sample is measured.
Fluorometry
More sensitive than colorimetry. Used to measure drugs, hormones.
Fluorometry
Atoms absorb light of specific wavelength and emit light of longer wavelength (lower energy)
Fluoremetry
COMPONENT PARTS:
Light source, primary monochromator, secondary monochromator, detector, readout device
Fluorometry
Light parts of Fluorometry
Mercury lamp
Xenon arc lamp
Doesn’t require excitation radiation or monochromator like Spectro, AAS, fluorometry. Extremely sensitive. Used for immunoassays.
Chemiluminiscence
Chemical reaction that produces light.
Chemiluminescence
Chemicluminiscence usually involves [4]
acridinium
esters
dioexerances
luminol
COMPONENT PARTS:
Reagent probes, photo multiplier tube, sample & reagent, cuvette, readout device
Chemiluminescence
Used to measure proteins in urine and CSF
Turbidimetry
Measures reduction in light transmission by particles in suspension; blocked light
Turbidimetry
Used to measure ag-ab reaction
Nephelometry
Similar to turbidimetry but light is measured at angle from light source
Nephelometry
COMPONENT PARTS:
Light source, collinator, monochromator, cuvette, photo detector, readout device
Nephelometry
The amount of light reflected by a solution dispensed onto a white granular surface is inversely proportional to the concentration of the sample
Spectrophotometry
Luminiscence is based on an energy exchange process that occurs when certain compounds absorb electromagnetic radiation, become excited and return to an energy level lower than or equal to their original level
Fluorometry
detects light scatterd at various angles
Nephelometry
detects reduction in light transmission due to particle formation
turbidimetry
When light passes from one medium to another, the light beam changes its direction at the boundary surface.
Refractometry
Based on the measuring changes in the colligative properties of solutions that occur owing to variations in particle concentration.
Osmometry
Measures multiple properties of cell suspended
Flow Cytometry
Measurement of the current or voltage generated by the activity of specific ions
Electrochemistry
Under electrochemistry
[5]
Potentiometry
Coulometry
Amperometry
Voltametry
Conductance
measurement of voltage between 2 electrodes in a solution
Potentiometry
measures the quantity of electricity in coulombs needed to convert an analyte to a dff. oxidation state
coulometry
measurement of current flow produced by an oxidation-reduction reaction
amperometry
measures a potential applied to an electrochemical cell and the resulting current
voltametry
measures the ability of a solution to carry an electrical current
Conductance
measurement is based on the change in electrical resistance across an aperture when a particle in conductive liquid passes through this aperture
impedance
potential difference bet 2 electrodes directly related to an analyte conc
ISE
___________separation of charrged particles in electrical field. Anions move to positively charges pole [anode]; cations to negatively charged pole [cathode]. The greater the charge the [faster/slower] the migration
Electrophoresis
faster
In protein electrophoresis,
the rate of migration depends on:
size
shape
molecule charge
In protein electrophoresis,
the support medium are
cellulose acetate
agarose
In protein electrophoresis,
the buffer used is the_______ at pH_____
Barbital buffer
8.6
In protein electrophoresis,
the stains used are [4]:
Coomassie brilliant blue
Ponceau S
Amido blue
Bromphenol blue
In protein electrophoresis,
the charge is?
proteins are negatively charged & move toward anode
In protein electrophoresis,
the order of migration [fastest to slowest]
albumin
alpha-1
alpha-2
beta
gamma
In protein electrophoresis,
the largest fraction is the?
albumin
In protein electrophoresis,
buffer flow toward cathode > causes gamma region to be cathodic to the point of application
Electroendosmosis
In protein electrophoresis,
__________they must be concentrated first because of low protein concentration.
Urine
CSF
___________ this type of proteins migrate gamma region here.
Bence Jones proteins
In protein electrophoresis,
it has a prealbumin band
CSF
Acute inflammation increases what?
alpha 1, 2
Chronic infection increases what?
alpha 1, 2, gamma
Cirrhosis increases what?
beta, gamma [polyclonal]
Monoclonal gammopathy
sharp in 1 immunoglobulin [gamma] “M” spike
Polyclonal gammopathy
diffuse increase in gamma
Hypogammaglobulinemia
decreased in gamma
Nephrotic syndrome
increased in alpha 2
decreased in albumin
Alpha-1 antitrypsin deficiency
decreased alpha-1
increased in beta OR
unusual band between alpha-2, beta
Hemolyzed spx
extra band [fibrinogen] between beta & gamma
plasma
continues to be one of the most rapidly advancing areas of lab medicine
Clinical chemistry
is a polygot discipline combining chemistry, biochemistry, immunochemistry, endocrinology, toxicology, anachem, informatics and doubtless other specialties; necessary to support to physicians and other healthcare providers to improve the diagnosis and treatment of px
Clinical chemistry
means that the multiple samples are tested in a ‘run’.
Batch analysis
means samples are tested one adter the other and results are reported in the same order
sequential analysis
________is a form of sequential analysis through a continuous system at a constant rate. E,g [1]
Continuous flow analysis
Autoanalyzer
each sample is tested in a seperate cuvette or other reaction chamber w reagents added to each individual sample container
Discrete analysis
uses analytical line
Single channel analysis
uses two or more lines or channels, each dedicated to a single test , and analysis occurs simultaneously.
Multiple-channel parallel analysis
spx are tested in or out of each sequence w/ each other, as reaction vessels are available and without regard to accessioning order, although testing of designated spx, such as STATS, may be given priority
random access analysis
are either endpoint tests [rxn is complete after a fixed time] or continuous monitoring tests [multiple data points recorded over a specified time interval]
Assays
Steps in automated analysis:
Sample ID used
usually by bar code reader
Steps in automated analysis:
Test selection used
LIS
In automated analysis,
Sampling used
closed-tube sampling from primary collection tubes
Some short sample + clot detection
Steps in automated analysis:
usually by syringes, pumps, or pressurized reagent bottles. Vitros uses ______slides.
Rgt delivery
dry
Steps in automated analysis:
Chemical reaction is through
Mixing & incubation
Steps in automated analysis:
Measurements used
Visible & UV spectrophotometry
ISE
Fluoresence polarization
Chemiluminiscence
Bioluminiscence
Automatic dilution
Steps in automated analysis:
Data handling
conc derived from calibration curve
Steps in automated analysis:
reporting
LIS through interface
Steps in automated analysis:
Troubleshooting
remotely by modem
inventor of continuous flow analyzers
Leonar Skeggs
In the 1956, Leonard Skeggs developed this first practical system for measuring urea, glucose, calcium
AutoAnalyzer
4 Main Types of automatic analyzer for processing clinical chemistry tests
Continuous flow system
Discrete Sampling Analyzers
Centrifugal Fast Analyzers
Thin-film Analyzers
all sample are carried through the same analysis pathway
Continuous flow system
all samples automatically pass from one step to another without waiting to bring the samples to the same stage of completion
Continuous flow system
the reactions are not necessarily carried to equilibrium since samples and standards are treated exactly alike.
all reactions must be carried out until equilibrium is reached
Continuous flow system
Discrete Sampling Analyzer
this system has won wide acceptance in both routine and reasearch laboratories
Continuous flow system
Examples of Continuous flow system
Technicon Autoanalyzer II
Sequential Multiple Analyzer [SMA 6/60]
SMA [12/60]
Sequential Multiple Analyzer w/ Computer [SMAC]
capable of running 3 different tests at 60-80 samples per hr
Technicon Autoanalyzer ii
capable of running 6 tests at 60 spx per hr
SMA 6/60
capable of running 12 tests at 60 samples pr hr
SMA 12/60
capable of running 40 tests at 120 samples per hr
SMAC
features of continuous flow system are the use of _______ of different diameters & __________ for continuous pumping of samples/rgts.
plastic tubes
peristaltic pump
In the introduction of air bubbles in continuous flow system, its purpose is to:
–separate sample & rgt streams into ________.
–[action] one sample from next
–for continuous _______ of tubing
– prevents ______________
segments
separate
scrubbing
cross-contamination
Each sample reactions is handled in separate compartment & does not come into contact w/ another sample.
Discrete sampling analyzer
The samples and standards are handled on a batch basis and must be brought before proceeding to the next procedure.
Discrete sampling analyzer
This system stimulates very closely manual procedures except that the various steps arre done automatically
Continuous flow analyzer
Discrete sampling analyzer
examples are:
Dupont ACA
American monitor KDA
Abbott ABA-100
ABA- 200
Beckman ASTRA 8/4
Beckman DSA 560/564
Biochromatic Analyzer
VP Analyzer
Discrete sampling analyzer
as the rotor is accelerated centrifugal force moves the reagents and sample to a mixing chamber and then through a small channel into the cuvette. As the filled cuvette rotates past a fixed light beam, the absorbance of the reaction is measured spectrophotometrically
Centrifugal Fast analyzers
Examples of Centrifugal Fast Analyzers
CentrifiChem
RotoChem
Thin film analyzers example
Kodak-Ektachem
developed by Norman Anderson
Centrifugal fast analyzers
A 16mm square chip which contains several thin layers, accepts a metered drop of serum, spreads it evenly int a reagent layer, then confines the colored product to a fixed area for reflectance spectrophotometry.
Thin-film Analyzers
