SCYM (ASCP) Quizlet (2025/2026) ACTUAL EXAM
COMPREHENSIVE QUESTIONS AND VERIFIED
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Most modern flow cytometers tightly position the
sample for optical analysis via hydrodynamic focusing.
Hydrodynamic Focusing Here, a carrier fluid called the sheath fluid is used to
position the sample of cells into a single file for
optical interrogation.
The central stream (sample stream) is focused and
surrounded by the secondary slower stream (sheath
Hydordynamic focusing fluid). The shape and size of the flow cell is crucial to
and sheath fluids hydrodynamic focusing, and traditionally the cell is
nozzle shaped. ... In a flow cytometer, the sheath fluid
pressure is constant while the sample fluid is adjusted
The difference between the sample pressure and the
Sample Pressure and the sheath pressure is the differential pressure. This
Sheath Pressure controls the width of the core stream and the total
number of cells passing the laser intercept.
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Differential pressure based flow cytometers currently
dominate the market. These systems have two
differential pressure based pressure regulators. The first is at a constant pressure
flow cytometers that sets how fast the fluids runs at. The second is
regulated by the investigator (like on this LSR-II
control panel).
Low differential pressure allows the cells to move past
the interrogation point one at a time. .... One kind
Generation of differential
involves generating pressure using a pump and
pressure (syringe pump,
regulator system ... Differential pressure based fluidic
pressure based)
system. ... peristaltic and/or syringe pumps to deliver
the sample into the instrument.
In syringe-pump-driven microfluidic systems, pressure
fluctuations are observed in an elastic microchannel.
Characterization of The syringe pump is driven by an electrical stepper
syringe-pump-driven motor, from which mechanical oscillations are
induced pressure expected to generate flow-rate fluctuations and in
turn leads to the pressure fluctuations in the channel
flow.
Filters are pieces of glass coated on both sides that
allow light of a certain collection, or band, of
Optical Filters wavelengths to pass through while absorbing or
interfering with photons of other wavelengths. These
come in bandpass, longpass, and shortpass flavors
A filter that allows light between a set wavelength to
pass through and reflects light above and below the
set wavelength. For example, a bandpass filter with a
Band Pass Optical Filter
wavelength of 550/40nm would allow light between
530nm and 570nm to pass through, but reflect light
below 530nm and above 570nm.
Longpass Filter wavelength above 650nM
Shortpass Filter wavelength below 488nM
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Dichroic mirrors can block light by phased reflection
allowing certain light to pass through and interfering
with other wavelengths. For example, a 500LP
dichroic mirror would transmit light above 500 nm and
reflect the light below 500 nm in a different direction.
dichroics mirrors
A 525SP dichroic mirror would transmit all light below
525 nm and reflect all light above 525 nm in a
different direction. These dichroic mirrors are critical
in the directing and capturing of light by the
detectors.
filter that reduces or modifies the intensity of all
neutral density filter wavelengths, or colors, of light equally, giving no
changes in hue of color rendition
Polarization of scatter and fluorescence signals in
flow cytometry. ... depending on the light source(s),
polarization filter
the optical layout, and the types of mirrors and filters
used.
The light source can be a laser, an arc lamp or even
an LED. Today, the majority of instruments use a laser.
Lasers illuminate the stream with coherent, focused
light source light of specific wavelength (energy) and power. This
illumination facilitates the generation of fluorescence
signals from cells labeled with fluorophores and light
scatter signals from redirected laser light.
Arc lamps need optical filters to select the
appropriate wavelength. They do not give the
arc lamp laser sensitivity needed to observe weak fluorescence but
offer a cheaper alternative for observing strong
fluorescences, for example, in DNA analysis.
Air-cooled argon-ion laser producing blue light at
488 nm. This wavelength is convenient for the
argon laser excitation of fluorescein, the first immunofluorescent
label to be used. Other air-cooled lasers in general
use include He-Ne (633 nm) and He-CD (325 nm).
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Solid state lasers producing light at 355, 405, 488,
530, 594, 635 and 780 nm are available. Most solid
solid state lasers
state lasers produce between 10 and 25 mW. There is
at least one diode laser giving 200 mW at 488 nm.
As the lasers interact with particles and cells at the
observation point or the interrogation point, scattered
lenses and fluorescence light is generated. In order to
measure this light, the cytometer needs to collect as
much of it as possible.
The optical collection system of a cytometer must
accomplish two goals. First, it must gather as much
What is the job of the light as possible from the interrogation point. Second,
lenses? it must collimate that light so that all rays propagate
parallel to each other and can travel through the
collection path without diverging.
Dichroic filters (sometimes called beam splitters) are
used in the flow cytometer at an angle often of 45°.
Short wavelength pass (SWP) filters transmit light
below a given wavelength and reflect light of longer
wavelengths. Long wavelength pass (LWP) filters work
Dichroic Filters in the reverse fashion. Their important parameters are
the wavelength for 50% transmission (the cut off for
LWP or the cut-on wavelength for SWP), the peak
transmission and the slope at the cut-on or cut-off
wavelength. Their properties depend on the angle at
which they are used.
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