LEC 14 Flashcards
def? natural fluorescence found within cells
autofluorescene
def? Any particle which generates signal after passing through the laser beam
event
what is forward scatter proportional to
cell size
what is side scatter proportional to
internal complexity like granularity, vacuoles, or inclusions
T or F: neutrophils will have a higher side scatter than lymphs and monos
T
def? plot that displays a single measurement parameter
histogram
def? plot that display 2 measurement parameters
dot plots
def? ID of cells in a population through the staining and ID of specific markers using monoclonal AB conjugated with fluorochromes
immunophenotyping
def? dye which allows ID of dead cells through a reduction in cell mmb integrity
viability dye
T or F: dead cells can be used in flowcytometry
F
what kind of cells can flowcytometery ID
subgroups
what kind of lymph has granules
large granule lymphs
on a dot plot with SSC vs. FSC: top left will have ____ granules and _____ size
increased, small
on a dot plot with SSC vs. FSC: bottom left will have ____ granules and _____ size
decreased, small
on a dot plot with SSC vs. FSC: top right will have ____ granules and _____ size
increased, big
on a dot plot with SSC vs. FSC: bottom right will have ____ granules and _____ size
decreased, big
what does it mean if a flowcytometer has more lasers
the more lasers the more advanced
routine blood diagnostics and blood count is still done by what today
Coulter counters
def? measuring the physical and chemical properties of cells flowing in a stream of fluid
flow cytometry
t or F: flow cytometry can measure other things besides cells
T
what does flow cytometery use to obtain info about a cell
- forward light scattered
- side light scattered
- fluorescence
what can flow cytometery tell you about leukemias
- morphology
- immunophenotype
- cytogenentics
what component of FC? sheath fluid tank and flow cell
fluidics
what component of FC? lasers and filters
optics
what component of FC? FSC, SSC, and FL detectors
detectors
what component of FC? ADC system and data processing system
electronics
def? allows cells to be interrogated one by one by the laser beam
hydrodynamic focusing
when will cells produce the max amount of optical signal
when the cell is in the center of the laser beam
what kind of culture must cells from solid tissue be in to use in FC
single cell suspension
how are whole blood and BM aspirates prepared for FC
must be anti-coagulated
how are tissue samples and BM biopsies prepared for FC
culture media
t or F: at least 85% of cells need to be alive for FC
T
T or F: cells are alive on a PBS
F
what are the 2 main objectives of data analysis in FC
- obtain statistics
- ID abnormal populations based on light scatter and staining patterns
what does CD stand for
cluster of differentiation
what kind of cell do CD molecules ID
leukocytes
what is used to detect CD markers
monoclonal ab and fluoresces
what are the cell markers for t helper cells
CD3+ CD4+
what are the cell markers for t cytotoxic cells
CD3+CD8+
what are the cell markers for b cells
CD19+CD20+CD3-
what are the cell markers for NK cells
CD16+CD56+CD3-
what are the cell markers for HSCs
CD20-CD25-CD34+
what is another name for scatter plots
bi-variate plots
what kind of marker is CD 45
pan-leukocyte marker
what is the order of strongest expression of CD 45 in WBCs from most to least
- lymphs
- monocytes
- granulocytes
t. or F: erythroid precursors are CD45 neg
T
the more shift to the ____ the more intense fluorescence signal
right
def? electronic boundary an operator uses to delineate cell clusters
gate
the dot plot with CD45/SS helps with detection of what
blast
what condition do CD55 and CD 59 belong to
PNH
t or f: if a sample is more than 48 hrs old it should not be analyzed using FC
T
what anti-coagulant is used on BM aspirates
heparin
