Liquid Samples - ANSWER Blood, spleen, bone marrow, fresh marine water
Adherent Samples - ANSWER Cell lines, some tumors
Solid Tissue Samples - ANSWER Tumors, organs, plants
Cons of enzymatic disruption methods - ANSWER Can strip surface epitopes
How to prevent coagulation - ANSWER ACD, EDTA, heparin, or physically disrupt cells us-
ing a method such as vortexing
How long can whole blood remain at room temperature? - ANSWER 24 hours, then re-
frigerate for up to 72 hours
Cells that are too large for flow cytometry - ANSWER Cardiomyocytes and Megakaryo-
cytes
Factors to consider when fixing to preserve the epitope - ANSWER Fixative type, fixation
time, temperature, fluorochrome compatibility, surface marker compatibility
Alcohol fixation cannot be used on these fluorochromes - ANSWER PE and GFP
Aldehydes - ANSWER formaldehyde, increases background, destroys epitopes, 4% per-
meabilizes
1
, Alcohol - ANSWER Ethanol and methanol permeabilizes, alters morphology, nuclear, cell
cycle
Harsh Detergents - ANSWER Triton X-100, NP-40
Gentle Detergents - ANSWER Saponin, Lysolecithin
Centrifugation Gradients - ANSWER Seperate samples based on density
Percoll
Ficoll
Do voltage plates push or pull droplets? - ANSWER Push. They are called "deflection"
plates.
Probe Types - ANSWER Antibodies, viability/DNA dyes, physiological, tracking dyes, fluo-
rescent proteins
Fluorochrome Selection - ANSWER Antigen density, protein, co-expression, optimal com-
bination, photostability, F/P ratio, quenching, signal to noise
Types of Assay Controls - ANSWER Fluorescence minus one, autofluorescence, biological
systems control, background measurement controls
Assay Optimization - ANSWER Appropriate use of limited sample, frequency of target, cell
concentration, kinetics, scalability, blocking, statistical design
Fluorochrome Issues - ANSWER Antigen density, protein co-expression, optimal combina-
tion, photostability, F/P ratio, spectral overlap, compensation, quenching
2
Adherent Samples - ANSWER Cell lines, some tumors
Solid Tissue Samples - ANSWER Tumors, organs, plants
Cons of enzymatic disruption methods - ANSWER Can strip surface epitopes
How to prevent coagulation - ANSWER ACD, EDTA, heparin, or physically disrupt cells us-
ing a method such as vortexing
How long can whole blood remain at room temperature? - ANSWER 24 hours, then re-
frigerate for up to 72 hours
Cells that are too large for flow cytometry - ANSWER Cardiomyocytes and Megakaryo-
cytes
Factors to consider when fixing to preserve the epitope - ANSWER Fixative type, fixation
time, temperature, fluorochrome compatibility, surface marker compatibility
Alcohol fixation cannot be used on these fluorochromes - ANSWER PE and GFP
Aldehydes - ANSWER formaldehyde, increases background, destroys epitopes, 4% per-
meabilizes
1
, Alcohol - ANSWER Ethanol and methanol permeabilizes, alters morphology, nuclear, cell
cycle
Harsh Detergents - ANSWER Triton X-100, NP-40
Gentle Detergents - ANSWER Saponin, Lysolecithin
Centrifugation Gradients - ANSWER Seperate samples based on density
Percoll
Ficoll
Do voltage plates push or pull droplets? - ANSWER Push. They are called "deflection"
plates.
Probe Types - ANSWER Antibodies, viability/DNA dyes, physiological, tracking dyes, fluo-
rescent proteins
Fluorochrome Selection - ANSWER Antigen density, protein, co-expression, optimal com-
bination, photostability, F/P ratio, quenching, signal to noise
Types of Assay Controls - ANSWER Fluorescence minus one, autofluorescence, biological
systems control, background measurement controls
Assay Optimization - ANSWER Appropriate use of limited sample, frequency of target, cell
concentration, kinetics, scalability, blocking, statistical design
Fluorochrome Issues - ANSWER Antigen density, protein co-expression, optimal combina-
tion, photostability, F/P ratio, spectral overlap, compensation, quenching
2
