1.10.24basic Flashcards
Centrifugal force depends on 3 variables…and separates based on…
mass, speed, radiusdensity
speed expressed as…force generated express as…or…angular velocity value…r=
revolutions/min(rpm)relative centrifugal force/RCF or gravity (g)1.118x10*-5r=radius in cm from center of axis to bottom
Equation for speed of centrifuge related to RCF/RCF=
1.118x10-5 x r x (rpm)’210 to neg 5*2=squared
CLSI
clinical and lab standards institute
CLSI recommend blood be centrifuged at an RCF of…or…for…min
1,000-1200xgOR3500 rpm for 10-15 min
…or… centrifuge where tubes are horizontal position when spinning and in a vertical position when stopped
Horizontal head or swinging bucket
…or…centrifuge where tubes are at angle in sample bucket, allowing small particles to sediment faster
fixed angle or angle head
Centrifuges that separate small liquid samples of <=2mL in plastic conical shaped containers w/integral snap cap
microcentrifuges
High speed centrifuge up to 150,000rpm to separate chylomicrons from serum
ultracentrifuge
strobe light determines…
speed
Check speed of centrifuge with…or….every…
tachometer or strobe light, every 6mths
NIST
national institute of standards and technology
Check time with…timer, every…
NIST traceable timer, 6mths
centrifuge used to sediment small particles faster
fixed angle/angle head
centrifuge used to separate chylomicrons from serum
ultra
Glassware/plasticware must fall into class…or….Institute that recommends tolerance limits…Preferred glassware class…Usually made of… …
Class A, BNIST recommendedPreferred is A and made of borosilicate glass
Pipette that is electronic, prepare multiple samples…Pipette that are handheld, disposable tips w/air displacement or positive displacement
Automatic pipettesemi-automated pipette
Mechanism of action which relies on a piston for creating suction to draw the sample into a disposable tip that’s changed after useA certain volume of air remains between the piston and the liquid.
air displacementie. pipettes used in lab
Mechanism of action which has a moving piston in pipepette tip or barrel much like a hypodermic syringe, doens’t require a different tip and is used as a dispenser.The piston is in direct contact with the liquid.
Positive displacmentie. osmo pipette
Which pipete requires recalibration…how often per year…Two methods used to calibrate…
automatic pipettes, 4x/yrgravimetric and volumetric
Calibration that verifies accuracy by the amt of water dispensed by pipette. Weight of water is proportional to the volume dispensed.Weight=volume
Gravimetric
Calibration that verifies accuracy of amt of dye thats pipetted into a specific volume of water and is read spectrophotometrically.absorbance=amount
volumetric
Which class of glassware doesn’t need to be recalibrated
A
Handheld that require bulb to apply suction
manual
…or… is glass pipette that has a bulb in the middle w/tapered delivery tipaccuracy is 1:….
transfer or volumetric1:1000
..or…Pipette thats uniform in diameter w/tapered delivery tip
measuring or graduated
Pipettte with two frosted bands
Last drop blown outb after pipette drains
Pipete must be rinsed w/solvent after first liquid has drained
To contain, TC
Pipette filled and allowed to drain by gravity. Held vertically, tip placed against side of vessel
To deliver, TD
Pipette filled, drained and remaining fluid in tip blown out, has two frosted bands at top
blow out
Pipette held in vertical position w/tip against vessel and drained by gravity
self draining, ie. TD
Organization that maintains standards for temps
NISTnational institute of standards and tech
Thermistor probe is also called
an electronic probe
3 types of thermometers
mercury/liquid in glassdigitalelectronic/themistor probes
Gold/platinum thermometer calibrated on international temp scale w/lead wires and sheath
NIST Standard ref material(SRM)
All NIST certified thermometers come with a…NIST traceable thermometers come with
NIST cert of calibrationNIST cert of traceability.certificate issued by manufacturer
Temp for incubators, heat blocks, water baths
36-38C
RT
20-24C
Lab fridge…BB fridge…
lab 2-8CBB 1-6C
Lab freezer…BB freezer…
lab <= -20CBB <=-65C
Sln can use as alternative to lens sln…sln not to use…
can use methanol, not xylene
Most common microscope, white light illuminates sample from below…limitation…
brightfieldlow contrast of biological samples
Microscopy improves contrast of unstained, transparent. Quantity of unscatted light/directly transmitted is minimized collecting only light scattered by sample… the field around the specimen (i.e., where there is no specimen to scatter the beam) is generally dark.Good to view…
Darkfieldspirochetes
Uses electrons to illuminate and creates enlarged image. Creates higher resolving power and higher magnification.Uses electrostatic and electromagnetic lenses. Focuses on a specific plane…Good to view…or… …
Electrontissue or tumor markers
Uses fluorescense. High energy light illuminates, emits light of lower frequency. Dyes used to stain structures…Good to view…
FluorescentANA
Small phase shifts in the light passing thru transparent specimen showing differences in refractive index as diff in contrast…An optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations.Good to view…
phase contrastmanual PLT cts
Only transmits polarized lightgood to view…
polarizedcrystals
Gives 3D image. brightfield can be modified into this. Two split beams of light are polarized and recombined by objective prism.Recombination gives 3D
Differential interference contrastDifferential Interference Contrast (DIC) is a phase contrast technique that allows transparent structures to be visualized by exploiting changes in refractive index. In contrast to conventional phase contrast techniques, which are based on changes in optical path length, DIC is based on the gradient of the optical path length (rate of change in wavefront shear). Steep gradients produce the high contrast and 3-D relief effect that is characteristic of DIC.
Magnification of:eyepiece..low power objective…high power objective…oil immersion objective…
eyepiece: 10Xlow: 10xhigh:40xoil: 100x
Knob to move slide right and left…knob to move stage forward and backward…
slide right/left: x axis motion knobstage forward/backward: y axis
Rotating disk under stage to vary intensity/size of cone of light
iris diaphram
Focuses light onto specimen
condenser lens
part that connects the eyepiece to the revolving nosepiece/lenses
tube
holds objective lenses
nosepiece
focuses light onto specimen, can move up and down
condenser lens
English units of measurement3 for length..1 mass..4 for volume…
length: inch,foot, yardmass: lbvolume: cup, pint, quart, gallon
Metric system is based on decimal system, multiples of tenlength…mass..volume…
length: metermass: gramvolume: liter
luminous intensity in SI
candela
International System of Units/SI worldwide standardized system, recommended by CLSILength:Mass:Time:amt sub:temp: electric current: luminous intensity:
Length: meterMass: kgTime: secamt sub: moletemp: Kelvinelectric current: ampereluminous intensity: candela
Deka
Deka…10*1
Hecto
Hecto…10*2
Kilo….
Kilo….10*3
Mega..
mega: 10 *6
giga
giga 10*9
Deci
Deci: 10*-1
centi..
Centi:10*-2
milli
milli: 10*-3
micro
micro: 10*-6
Nano:
Nano: 10*-9
pico
pico: 10*-12
femto
femto: 10*-15
Celcius from farenheit
5/9x (F-32)
Farenheit from Celcius
(9/5)C +32
Kelvin from Celsius
C+273
95F to C
5/9x(95-32)=35C
95C to F
(9/5x95)+32=203F
95C to Kelvin
95+273=368K
BUN/Creatinine ratioBUN 24.0Creatinine 2.4Give ratio
10:1
CHOL/HDL ratioTC 200HDL 40
5:1
Which cell lines aren’t included in the Myeloid part of M:E calculationWhich cell line is included in Erythroid ratio
Lymphs and MonosNormoblasts
Write fraction, 1 pt solute to 1 part diluent
1/22 is total volume
pt:pt is
pt analyze + total parts (including diluent)
1:1 dilution=1:21:5
1 pt serum+0 pt diluent1:2 1 pt serum + 1 pt diluent1:5 1 pt serum + 4 pt diluent
1:2, 1:4, 1:8 etc is a 1:5, 1:25, 1:125 is a 1:10, 1:100 is a
two fold, x2five fold, x5ten fold x 10
5% w/w sln equals how much solute to total solutionHow much diluent is added
5g solute per 100g total solution95g diluent added to 5g making 100g sln
Make 100g of 5% w/w sln of NaCl
5g/100g is 5% w/w,5/100x 100g wanted=need 5g NaCl to 95g diluent to make a total of 100g sln
10% w/v sln equals how much solute to total sln
10g of solute to 100mL of total sln
Make 1,000 ml of 10% w/v of NaOH
10% w/v = 10g/100mlx 1000mL= 100g of NaOH to add to 900ml diluent to get 1000mL total
2% v/v equals how much solute to sln
2ml/100ml
Make 50mL of a 2% v/v of HCl
2% v/v = 2ml/100mlx50ml=1ml of HCl added to 49ml diluent equals total of 50ml
Molarity/M
Have g/L, covert to mole/L by get gmwg/L x mole/L= M
40g NaOH/L, what is molarity?
1 mole NaOH is 40 GMW,40g/L x 1 mole/40 =1mol/L=1M
Nomality (N)
N=Equivalent/L1 Equivalent=gmw/valenceValence is # of H
Osmolality=mOsmol/kg of H2ONormal range…calculation…
Normal range 275-295 mOsm/kg1.86(NA) + GLU/18 + BUN/2.8
Osmolality of NA 150Glu 180BUN 14
294 mOsmol/kg H2)
Osmolar gap
measured-calculated
Concentrated SlnsC1V1=C2V21 is, 2 is
1 is known2 is unknown
PT has 350 PLT ct, how many mL of it needed to get 7mL of with concentration of 250
350x?mL=250x7=5mL
Ratio of SD of a distribution to its arithmetic mean.Used to express precision and reproducibility of an assay
Coefficient of Variation, CV
Coefficient of Variation, CV used to express…and…Acceptable CV is…
Used to express precision and reproducibility of an assayLess than 5% for most analytical systems
/X is
mean
CV formulaMean is 200, SD is 5, calculate CV
SD/Mean x 1005/200 x 100 =2.5%
Gives an estimated degree of uncertainty. Probability of estimated range…expressed as..
Confidence intervalpercentage
Mean +- 1 SD=Mean +- 2 SD=Mean +- 3 SD=
1SD 68%2SD 95%3SD 99%
Given mean (/X) is 9.8, 1 SD is 0.2, calculate 95% confidence limits for control run with mean +- 2SD
1SD= 9.8 +- 0.22SD=9.8 +- 0.4, 9.4-10.2
pH formula
pH=pKa +log (base)(acid) (salt)(acid) (HCO3)(H2CO3)
Diff between cations and anions…cations..anions…calculation
anion gapcations: Na, K anions: Cl, CO2Na+K - Cl+CO2
Law that states concentration of sub is directly proportional to amt of radiant energy absorbed or inversely proportional to the logarithm of the transmitted radiant energy2 formulas:proportional to absorbedversusindirectly proportional to transmitted…
Beers LawA=abcA=2-log%T
Formula for known/unknown absorbance…
Concentration of unknown=AbsUn/AbsStd x ConcenStd
Abs of 6.0g std is 0.40, abs of unknown is 0.350, concentration of unknown?
0.350/0.40 x 6.05.25g
Chain of custody things needed…
PT consent, security seal, to addresee onlystore in locked area
Examples of chain of custody
alcohol levels for DUIDOT drug testingpaternity, DNARapeMedical examiner
Digital version of patient medical history/date
EMRelectronic medical record
Series of computer network that interconnects computers in a limited area like hospital, using network media
LANlocal area network
A computer network that interconnects computers in a broad area such as international, using private or public network trainsports
WANwide area network
Comprehenstive integrated info system to manage medical, admin, financial, and legal aspects of a hospital/services
HISHospital info system
Computer software providing services to software apps beyond those available to the operating systemSoftware that lies between an operating system and the applications running on it. Essentially functioning as hidden translation layer, enables communication and data management for distributed applications.
Middleware
Computerized process for healthcare providers to electronically enter and order medical procedures/lab tests based on predetermined logarithm
CPOEcomputerized provider order entry
CLIA stands for
clinical laboratory improvement amendments
CDC stands for
centers for disease control
CMS stands for
centers for medicare/medicaid services
Program that establishes quality standards for all labs…Published by…and…
CLIAplublished by CDC and CMS
Organization that categorizes and grades testing based on the complexityof testing
FDA
Two criteria to be waived test
cleared by FDA for home usesimple/accurate as to pose no reasonable risk if performed incorrectly
Examples of waived
blood glucose/cholesterolurine prego, dipstick/tablet urinalysisovulationnonautomated ESRspun HCTHmg copper sulfateHmg by single analyze instruments
Who determines if a test meets criteria for being a waived test
HHSDept of Health and Human Services
Complexity?-manual procedures w/limited steps-microscopy of urine sediment-automated heme w/out diff-primary culture set up-urine culture/colony ct kits-gram
moderate
To be categorized as PPM, provider performed microscopy, must meet criteria
-physician, supervised midlevel practioner, dentist-moderately complex-main instrument is a microscope-control materials not available-specimen is labile, delay would affect results-need limited handling/processing
Examples of PPM tests
-direct wet mt preps, qualitative semen-KOH preps-pinworm, fern-postcoital direct, qualitative-urine sediment-nasal smears wbc, fecal wbc
3 criteria for high complexicity
-any modified FDA cleared test-lab developed test/LDT-cytology
Complexity?-serogrouping/typing-manual procecures w/multiple steps/reagent prep-concentration/diff staining-antimicrobial testing-gel immunochemical -electrophoresis, flow-pap smear
high
These two can conduct an inspection of any accredited lab that’s issued a certificate of accreditation…How often CLIA requires…
CMS, dept of HHS2yrs
External quality control to verify accuracy of lab test methods
PTproficiency testing
Proficiency testing is required by… under…Usually done …times per yearNot required for…
CMS, under CLIA3 times per yearnot required for waived tests
Approves the PT program that must be enrolled in
HHS
Must maintain copy of PT related forms for minimum of….from the date of PT event
2 yrs
Satisfactory performance on PT testing is…for each analyzeexcept with immunoheme, some are..
80%100%
Unsatisfactory PT
same analyte in 2 consecutiveOR2 out of 3 testing events
Alternate/Split sample testing can be done for PT if…
approved PT isn’t availableSPLIT SAMPLES w/other or ref lab, established in house method, clinical validation by chart review, participation in ungraded/educational PT program
Examples of pre analytical testing
lab req, id, collection, storage, transportation, specimen adequacy, aliquoting, centrifugation
Oral requests can be made if…and within…
followed by written authorizationwithin 30 days
Standard that requires the use of at least 2 pt ids…3 examples of ids
Joint CommisionNational Patient Safety Standard-name, bday, med rec
How long keep records of test reqs/authorizations (can be patients chart/medical record)…Who may request records…
2 yrsHHS
Examples of analytical testing
reagents, calibration, QC, tempsrecording resultsverify normal rangecompetency of personell
Process of testing and adjusting an instrument/kit to provide a known relationship between the measurement response and the value of the substance being measured by the test
calibration
Records of patient testing kept for at least…BB records kept for…Blood/products…Pathology kept for..
-PT 2yrs-BB 5yrs-Blood/products: 5yrs or 6mths after exp which ever is later-path 10yrs
How long must a copy of each procedure w/dates and initial use/discontinuance be kept
2 yrs after discontinuence
Post analytic examples
verifying calculationsreporting resultsID/notify panic filingdata retrieval
Most errors occur in what two phases…
pre and post
percentage of error:pre…analytical..post…
pre 46%analytical 8%post 47%
Term used to describe monitoring/evaluating all aspects of testing and all phases.Uses the QC data and conducted over a period of time
QA
Term that is one component of QA, involves mostly the analytical phase, ensures accuracy, and performed daily
QC
standard that where concentration is determined by dissolving weighted amt of std in solvent and diluting to a stated volume/mass.Accuracy based on purity of std/solvent not analytical method.
primary calibration std
Standard where concentration is determined by an analytical method of stated reliability. Prepared from pure substances of known composition.
Secondary std
Process that sets the instrument to a specific parameter, also called standards
calibration
Substances that are patient like used to validate run after calibration and tests the accuracy and precision of the methodCan use sera/BF thats stable for at least…
controlsa year
Control reqs
should be pt likerun at lease 2 levelsstablenever used to calibrate
Term that is the agreement between the measured quanitity and its true valueHow close the value is to the true value
accuracy
Material prepared/purchased w/stated or expected value
assay control
Material whose value is not known except when assayed w/in lab by repeated testing
unassayed control
Aggrement among replicate measurements. How close the single values are to one another. The repeatability or reproducibility
precision
The more precise the method, the…the SD among replicate results
smaller
Actual middle value
median
value that occurs the greatest frequency
mode
average of all values, included in control material, represented by /x
mean
dispersion of control values about the mean.repeatability, reproducibility, precision
SD
If data have a normal distrubution, the mean/mode/median are…
about the same
Difference between highest and lowest values
range
average squared distance of data points from the mean
variance
square root of the variance
SD
SD expressed as percentage of the mean
CVcoefficient of variation
CV formula….acceptable CV is less than…% for most analytical systems
SD/mean x 100
1SD…% will fall to the right and left of the mean, so…% of all data fall within 1SD
34.1% fall to right and left68.2% of all data fall within 1SD
2SD…% of all data will fall between 2 SD
95.5% of all data fall within 2SD
3SD…% of all data will fall within 3SD
99.6% of all data fall within 3SD
The lab normally runs within …or…% confidence limit to be considered accurate and precise
2SD, 95%
A chart with a normal distrubution curve on its side. Used to determine if a run is acceptable/quality of run.Values within…on each side are accepted as being accurate and preciseAccurate test results fall w/in acceptable limits of +/-…SDs….% of the time
Levey Jennings chart2SD
Signal of errors…consecutive results fall on…side of mean…consecutive results fall outside……result falls outside….
5 consecutive results fall on 1 side of mean2 consecutive results fall outside 2SD1 result falls outside 3SD
6 steps in finding source of signal errors
- clerical2. reagent3. procedural/maintainence4. retest ctrl5. recalibrate instrument/new ctrl6. call tech
Error thats unpredicable, no obvious pattern and almost always preventable.Large deviation from mean.
Random error
3 Random error causes…Remedy…
operator error (pipetting etc)electronic fluctuation in instrumentinterfering substancesRemedy: repeat test with fresh reagents, good working pipettes
Error that happens over a period of time, data points are not evenly distributed about the mean.This can be described as…or….
Systemetic errorshift or trends