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Flow cytometry Exam Comprehensive Questions (Frequently Tested) and Complete Solutions Graded A+

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Flow cytometry Exam Comprehensive Questions (Frequently Tested) and Complete Solutions Graded A+

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Flow cytometry Exam
Comprehensive Questions
(Frequently Tested) and Complete
Solutions Graded A+
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Hydrodynamic Focusing - Answer: - when two streams of fluids
with different flow rates are running side-by-side and in the same
direction into a flow cell, then a laminar flow is created.

- The central stream (sample stream) is focused and surrounded
by the secondary slower stream (sheath fluid).

,- In a flow cytometer, the sheath fluid pressure is constant while
the sample fluid is adjusted.

- Hydrodynamic focusing properly aligns your cells, one by one, at
the junction where the analysis by lasers begins.



Generation of Differential Pressures: syringe pump - Answer: -
samples are loaded into holding lines that are then rapidly driven
through a simple mixer into a delay line.

- The mixed sample is then driven by drive syringe at a controlled
rate into the flow cell.

- This process requires high-speed valves to switch the delivery
from waste to the flow cytometer just after mixing. (This is a
critical step for hydrodynamic systems, as the high fluidic rates
that occur during mixing are incompatible with laminar flow.)



Generation of Differential Pressures: pressure based - Answer: -
In this method of sample delivery, a pressure differential is
maintained across the reservoirs containing the sample and
sheath fluid

- Once the pressure of the sample has increased above that of the
system, the sample is driven into the system.

- In these systems, the rate of sample delivery depends upon the
pressure gradient as well as the pressure drop that occurs along
the length of the capillary tubing.

, - are subject to pressure variations over time



Optical Filter: Long Pass - Answer: transmits light above a certain
wavelength



Forward Scatter - Answer: - Light that is scattered in the forward
direction after interacting with a particle, typically up to 20o offset
from the laser beam's axis, is collected by a photomultiplier tube
(PMT) or photodiode

- can give an estimation of a particle's size with larger particles
refracting more light than smaller particles, but this can depend
on several factors:

- the sample

- wavelenght of the laser

- collection angle



Side Scatter - Answer: - Light measured at a 90o angle to the
excitation line

- can provide information about the relative complexity (for
example, granularity and internal structures) of a cell or particle;
however as with forward scatter this can depend on various
factors

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