Below is a concise list of publications that may be useful when entering the small particle flow cytometry field.
First demonstration of quantitatively calculating light scatter
resolution using light scatter modelling.
de Rond L, Coumans F A W, Welsh J A, Nieuwland R, Leeuwen T G.,
van der Pol E,
Quantification of light scattering detection efficiency and
background in flow cytometry., Cytometry Part A, doi: 10.1002/cyto.a.24243 [Website]
Review outlining considerations when measuring and reporting EV
characteristics of size, concentration, refractive index, and
epitope abundance.
Welsh J A, van der Pol E, Bettin B A, Carter D R F, Hendrix A,
Lenassi M, Langlois M, Llorente A, van de Nes A S, Nieuwland R,
Tang V, Wang L, Witwer K W, Jones J C,
Towards defining reference materials for extracellular vesicle
refractive index, epitope abundance, size, and concentration., Journal of
Extracellular Vesicles, doi:
10.1080/20013078.2020.1816641 [Website]
Position paper providing single EV flow cytometry reporting
framework and minimum experimental guidelines.
Welsh J A, van der Pol E, Arkesteijn G, Bremer M, Brisson A,
Coumans F, Dignat-George F, Duggan E, Ghiran I, Giebel B, Görgens
A, Hendrix A, Lacroix R, Lannigan J, Libregts S, Lozano-Andrés E,
Morales-Kastresana A, Robert S, de Rond L, Tertel T, Tigges J, de
Wever O, Yan X, Nieuwland R, Wauben M, Nolan J, Jones, J C.,
MIFlowCyt-EV: a framework for standardized reporting of
extracellular vesicle flow cytometry experiments, Journal of Extracellular Vesicles.
doi:
10.1080/20013078.2020.1713526 [website]
First cross-platform comparison of fluorescence, light scatter,
and concentration calibration showing its utility across
instrument settings and different flow cytometers.
Welsh J A, Jones J.C. Tang V A.,
Light scatter and fluorescence calibration allow standard
comparisons of small particle data between different
instruments,
BioRxivs, doi: 10.1101/796961. [website] [cytometry files]
Outlines the characteristics of polystyrene, silica, and EV
particle light scattering. First release of FCMPASS
software.
Welsh J A, Horak P, Wilkinson J S, Ford V, Jones J C, Smith D C,
Holloway J A, Englyst N A,
FCMPASS software aids extracellular vesicle light
scatter standardisation., Cytometry Part A, DOI: 10.1002/cyto.a.23782 [website]
First EV flow cytometry standardization study to use light
scatter calibration.
van der Pol E., Sturk A., van Leeuwen T., Nieuwland R., Coumans
F., ISTH‐SSC‐VB Working group
Standardization of extracellular vesicle measurements by flow
cytometry through vesicle diameter approximation.
J Thromb Haemost 2018; 16: 1236– 1245, DOI: 10.1111/jth.14009 [website]
Paper outlining the options and consideration when performing
light scatter calibration.
de Rond, L., Coumans, F. A., Nieuwland, R., van Leeuwen, T. G., &
van der Pol, E.
Deriving extracellular vesicle size from scatter intensities
measured by flow cytometry
Current Protocols in Cytometry, 86, e43. doi: 10.1002/cpcy.43 [website]
Paper demonstrating the use of refractive index and sizing of
EVs using the ratio of forward and side light scatter
measurements.
van der Pol E., de Rond L., Coumans F. A.W., Gool E L., Böing A.
N., Sturk A, Nieuwland R, van Leeuwen T.G.,
Absolute sizing and label-free identification of extracellular
vesicles by flow cytometry
Nanomedicine, doi: 10.1016/j.nano.2017.12.012 [website]
This is the second installment of the MISEV guidelines and is
useful for knowing the nomenclature and orthogonal assays
expected of EV studies.
Théry C., Witwer K., et al.,
Minimal information for studies of extracellular vesicles 2018
(MISEV2018): a position statement of the International Society
for Extracellular Vesicles and update of the MISEV2014
guidelines
, Journal of Extracellular Vesicles, 8:1, 1535750, doi:
10.1080/20013078.2018.1535750. [website]
Short review on EV flow cytometry pitched at a beginner
level.
Welsh J.A., Holloway J.A., Wilkinson J.S., Englyst E.A.,
Extracellular Vesicle Flow Cytometry Analysis and
Standardization.
Frontiers in cell and developmental biology. 2017, DOI:
10.3389/fcell.2017.00078 [website]
Though pitched at a cell flow cytometry audience, this is a
useful manuscript for understanding flow cytometry nomenclautre
and the importance of characterizing fluorescence
sensitivity.
Wang, L. and Hoffman, R.A. 2017.
Standardization, calibration, and control in flow cytometry.
Curr. Protoc. Cytom. 79:1.3.1-1.3.27. DOI: 10.1002/cpcy.14 [website]
Thorough review on EV flow cytometry pitched at a beginner
level.
Nolan J.P.,
Flow Cytometry of Extracellular Vesicles: Potential, Pitfalls,
and Prospects, Current Protocols in Cytometry, 2015, DOI:
10.1002/0471142956.cy1314s73 [website]
First paper demonstrating EV diameter derivation from membrane
intercalating dye and first EV paper using fluorescence
calibration.
Stoner S.A., Duggan E., Condello D., Guerrero A., Turk J.R.,
Narayanan P. K., Nolan J.P.
High sensitivity flow cytometry of membrane vesicles.
Cytometry. Part A : the journal of the International Society for
Analytical Cytology. 2015, DOI: 10.1002/cyto.a.22787 [website]
Paper demonstrating the cross-platform sensitivity and
resolution differences when analyzing EVs.
van der Pol E., Coumans F. A. W., Grootemaat A. E., Gardiner C.,
Sargent I. L., Harrison P., Sturk A., van Leeuwen T. G. ,
Nieuwland R.,
Particle size distribution of exosomes and microvesicles
determined by transmission electron microscopy, flow cytometry,
nanoparticle tracking analysis, and resistive pulse sensing.
2014, DOI: 10.1111/jth.12602 [website]
Standardization study demonstrating the utility of Q&b for cell
analysis.
Perfetto, S. P., Chattopadhyay, P. K., Wood, J. , Nguyen, R. ,
Ambrozak, D. , Hill, J. P. and Roederer, M.,
Q and B values are critical measurements required for
inter‐instrument standardization and development of multicolor
flow cytometry staining panels.
Cytometry, 85: 1037-1048. doi:10.1002/cyto.a.22579 [website]
First paper to demonstrate swarm detection and perform EV light
scatter calibration.
van der Pol E., van Gemert M. J. C., Sturk A., Nieuwland R., T. G.
Van Leeuwen
Single vs. swarm detection of microparticles and exosomes by
flow cytometry, Journal of Thrombosis and Haemostasis. 2012, DOI:
10.1111/j.1538-7836.2012.04683.x [website]
ISAC position paper outlining the minimum requirements for
reporting a flow cytometry experiemnt. While predominantly
developed for cellular analysis many of the criteria are
applicable to small particle analysis. This framework was
tailored to small particle analysis in the MIFlowCyt-EV
framework, 2020.
Lee JA, Spidlen J, Boyce K, Cai J, Crosbie N, Dalphin M, Furlong
J, Gasparetto M, Goldberg M, Goralczyk EM, Hyun B, Jansen K,
Kollmann T, Kong M, Leif R, McWeeney S, Moloshok TD, Moore W,
Nolan G, Nolan J, Nikolich-Zugich J, Parrish D, Purcell B, Qian Y,
Selvaraj B, Smith C, Tchuvatkina O, Wertheimer A, Wilkinson P,
Wilson C, Wood J, Zigon R; International Society for Advancement
of Cytometry Data Standards Task Force, Scheuermann RH, Brinkman
RR.
MIFlowCyt: the minimum information about a Flow Cytometry
Experiment.
Cytometry A. 2008 Oct;73(10):926-30. doi: 10.1002/cyto.a.20623.
[website]
Paper outlining how MESF values are assigned.
Gaigalas AK, Wang L, Schwartz A, Marti GE, Vogt RF Jr.
Quantitating Fluorescence Intensity From Fluorophore:
Assignment of MESF Values.
J Res Natl Inst Stand Technol. 2005;110(2):101–114. Published 2005
Apr 1. doi:10.6028/jres.110.010 [website]