Laboratory Flashcards
Side scatter detects _______ light
refracted
Forward scatter detects _______ light
diffracted
FITC emits green light at _______ nm
525
PE emits orange light at _____ nm
575
PerCP emits red light at _____ nm
677
APC emits red light at _______ nm
660
Photo Multiplier Tubes (PMT) detect
Side scatter and fluorescence
Photodiodes detect
Forward scatter
Optical filter that transmits wavelengths above a specified cut-off
long pass
Optical filter that transmits wavelengths below a specified cut-off
short pass
Optical filter that transmits wavelengths in a narrow range around a specified wavelength
band pass
FL1 channel measures light emitted from this fluorophore
FITC (530nm)
FL2 channel measures light emitted from this fluorophore
PE (585nm)
FL3 channel measures light emitted from this fluorophore
PerCP (670nm)
Channel values appear linear but are actually _______ values; and the relationship between channels is _______, not additive
log; proportional
Channels are used for binning _______ values
linear
MESF bead calibration converts channel values to ______ scale
linear
Delta MCF increases with ______ incubation temp; ______ serum volume, and ________ cell number
increased; increased; decreased
Ficoll isolation cell purity
25-80%
Negative selection by Easy Sep cell purity
91 - 98%
6 elements of competancy
- Direct observation of test performance
- Recording and Reporting
- Record review
- Direct observation of maintenance
- Test Performance
- Problem solving skills
Break even point formula
BEP= Fixed costs/(Test price - Variable costs)
Relative Risk formula
[a/(a+b)] / [c/(c+d)]
Odds ratio
ad/bc
sensitivity
a/(a+c)
specificity
d/(d+b)
SSO will use _______ conditions for hybridization and _____ conditions for washing
non-stringent/stringent
42 CFR part 493
CLIA
45 CFR part 160
HIPPA
29 CFT part 1910
OSHA
Certificate of accreditation is good how long?
2 years
CMS can perform random inspections but will give lab _____ advanced notice
2 weeks
Changes in name, ownership, location or director of lab must be reported to _____ and _______ within _______.
CMS/ASHI/30days
Off site archived records must be retrievable within _____
48 hours
Competency must be performed ________ during first year and ______ thereafter. Also must be completed with ________ changes. Competency is not required for _______ unless they perform tests.
semiannually/annually/instrument or method/director
A new director must review and sign all procedures within ______
6 months
Cell control panel used for specificity validation must have 3 controls
- Positive for antigen
- Negative for antigen
- Positive for corrective antigen
Computer systems must have assessment performed this often
1 year
Calibration performed every _______ and during certain situations (name 3)
6 months
- Change in reagents
- Major maintenance or replacement of parts
- Controls are out of limits and other trouble shooting has failed
Lab must verify and document thermal cycler temperature accuracy this often
6 months
Continuing education requirement of ARB by ASHI: Director
50 hrs/year
Continuing education requirement of ARB by ASHI: General supervisor
27 hrs/year
Continuing education requirement of ARB by ASHI: Tech
12 hrs/year
Continuing education requirement of ABHI: CHA
60 hrs/3 yrs
Continuing education requirement of ABHI: CHT
60 hrs/3yrs
Continuing education requirement of ABHI: CHS
80 hrs/3yrs
Continuing education requirement of ABHI: Director
150 hrs/3yrs
Education requirements for Tech (3)
- Medical doctor or doctoral, masters, or bachelor’s degree in science
- Associate degree in lab science or medical technology
- 60 semester hours of which 24 hours should be medical laboratory technologist school OR 24 hours of basic science course (of which 6 hrs are chem, 6 hrs are bio, and 12 are miscellaneous science courses)
Methionine is the ______ codon
start
Methionine codon
AUG (ATG)
Stop codons
UGA (TGA), UAA (TAA), UAG (TAG)
Flow cytometry: must run _______ and _______ standards each time instrument is ________, _________, ______
optical/fluorescent/turned on/maintained/adjusted or fix due to problem
Controls must be run this often
daily
Controls for quantitative assays
2 controls of different concentrations
Controls for qualitative assays
Negative and Positive control
New lots: Parallel testing with previous lot must be performed; the number of tests to run in parallel is determined by ________
technical supervisor
Cell culture: Laminar flow hood must be used for cultures incubating _______
> 18 hrs
Cell culture: Incubators must be monitor for _____, ______, ______
temperature/CO2 concentration/humidity
MLC must have these controls
- Negative control for each responder cell consisting of responder cells stimulated by autologous cells
- Positive control for responder cell proliferation consisting of known HLA Class II disparate stimulator cells from unrelated donors as well as pooled donors.
Serum used in MCL culture medium should be screened for these 3 things
- Ability to support cellular proliferation
- Lack of cytotoxic antibodies
- Sterility
SSP must have these controls
- Internal control to detect technical failure
2. Negative control to detect contamination
NGS: ________ must be monitored by rotating control samples with different barcode sequences
fidelity
All UNOS histocompatibility labs must review and verify UNET data for each patient and documentation of this review must be maintained for at least _______
3 yrs
STR: If subpopulations are analyzed, ______ of each cell subset must be obtained and documented
purity
Reports must include at minimum
- Lab name
- Director’s name
- Lab address
- CLIA/ASHI #
- Date of collection
- Date of report
- Unique patient identifier
- Method
- Result
There must be documentation of condition and disposition of ________ specimens
rejected
Parallel testing between instruments, sites, and methods must be performed this often
q 6 months.
Approximate length of Class II amplicon
270 bp
Approximate length of Class I amplicon
1000 bp
Requirements for general supervisor (5)
- lab director
- technical supervisor
- Medical doctor degree + 3 ys of HLA lab experience
- Doctoral, masters, bachelor’s degree in science + 3 yrs of HLA lab experience
- Served as General Supervisor before 2/28/92 and functioned as technologist + 6 years of lab experience, 2 of which in HLA lab
NFPA Diamond: _____ red square is for ________
Top/Fire hazard (flash point - will not burn to <73F)
NFPA Diamond: _____ yellow square is for ________
Right/Reactivity (volatility - stable through may detonate)
NFPA Diamond: _____ white square is for ________
Bottom/Specific hazard (e.g., oxidizer, acid, alkali, corrosive, radiation)
NFPA Diamond: _____ blue square is for ________
Left/Health Hazard (Normal to Deadly)
2 methods used in screening for Class II antibodies using cell based assays
- Parallel screening using B and T cell targets
2. Absorb out Class I antibodies with platelets
MLC: Responses considered non-stimulatory if stimulation index is _______
<2.5
Stimulation index formula
(A/Bx)/(A/Ax) [Response to allogeneic stimulator cell/Response to autologous stimulator cell]
MLC: Patients with ________ can have unresponsive cells making test uninterpretable
leukemia
Best statistical method for evaluation of serologic typing reagents
correlation coefficient
Optimizing _______ can prevent plateau effect in PCR
number of cycles
CSFE is a _________ dye
supravital
trypan blue is a _______ dye
vital
Thawing solutions without mixing can result in _________
solute gradients
EBV transformed B-cells for HLA testing can result in ________ due to ________.
cytotoxicity/false positive; non-HLA antibodies
Probes in SSO generally bind membrane distal portion of HLA molecules, which refers to ________
top, exposed portion of molecule (i.e., the peptide binding portion, where the polymorphisms lie)
HCFA stands for
Healthcare Finance Administration
Anti-T cell therapy (ATG) can cause _______ on antibody assessment
false Class I reactivity
__________ areas should be physically separated in oder to prevent DNA contamination
Pre-amplification
AMOS XM introduces __________ prior to complement to prevent ___________ factors
wash steps; anticomplementary
_________ is considered to be the least sensitive XM method
Standard NIH
Selection of sera for XM of allosensitized patients should be based on ________ and ___________
historic and current sensitizing events.
Medicare Part A covers reimbursement for HLA testing during __________ period
pre-transplantation
_________ annealing temperature can ________ specificity in allele-specific PCR
Increasing/increasing
Low control in lymphotoxicity assay can be the result of _______ contamination or _______ control serum
T-cell/out-dated
MLC: Stimulator cells are irradiated with _______ rads to inhibit ________ but preserve _______
3000 - 4000/DNA synthesis/Class II expression
MLC: increase reactivity of autologous control can be caused by _______, ________, and ______
viral infections, recent transfusions, leukemia relapse
One-way primary MLC is basically a ___________
cellular XM procedure
Methods for HLA antigen isolation/purification
detergent solubilization and affinity chromatography
MLC: limiting dilution analysis calculates frequency of functional lymphocyte populations based on assumption that those populations follow a _________ distribution
Poisson
Isolating of peptides from HLA molecules can be performed after isolating the HLA molecule by _______ of peptides
acid elution