Flow sorting a brief overview Ian Titley Haematology Oncology Leukaemia Research Fund Institute of C

1
Flow sorting a brief overview  Ian
TitleyHaematology OncologyLeukaemia Research
FundInstitute of Cancer ResearchLondon
2
Flow sorting - a form of fluorescence microscopy
whereby single cells in liquid suspension can be
identified and physically separated from each
other according to unique characteristics
3
History of cell sorter 1st prototype at Los
Alamos National Laboratory (LANL) in 1965
developed by Mack J Fulwyler by joining Coulter
volume sensing (invented by Wallace and Joseph
Coulter) with the newly invented ink-jet printer
technology. Biologist Leonard Herzenberg at
Stanford University was the first to recognise
the utility for this technology to biological
applications. With colleagues from the Genetics
Dept Instrument Research group and using
Fulwylers plans developed a machine to sort
fluorescently labelled cells. The 1969 instrument
had a mercury arc lamp as light source and in
1972 an argon ion laser. Funding from the
National Institutes of Health (NIH) allowed
Herzenberg and the Stanford engineers to interest
the medical products company Becton Dickinson
(BD) to convert their prototypes into the first
commercial instruments, the FACS (Fluorescence
Activated Cell Sorter) in 1975. Smithsonian
Institute Archives Video History
Collection http//www.si.edu/archives/ihd/videocat
alog/9554.htm
4
Electrostatic  Mechanical  Lab-on-chip
5
http//www.cardiff.ac.uk/medicine/haematology/cyto
netuk/introduction_to_fcm/cell_sorting.htm
6
BD VantageSE DiVa option
Beckman Coulter Epics Altra Hypersort
DakoCytomation MoFlo
BD FACSAria
7
(No Transcript)
8
Sorting into a 96 well plate (Taken from BD
FACSAria brochure)
9
(No Transcript)
10
Electrostatic Pros Cons Fast
(?x104cellssec-1) Expensive to buy and
run Multiparameter Needs trained operator High
purity Aerosol biohazard 4-way
11
Electrostatic  Mechanical  Lab-on-chip
12
http//www.cardiff.ac.uk/medicine/haematology/cyto
netuk/introduction_to_fcm/cell_sorting.htm
13
Partec PPCS

Union Biometrica COPAS
BD FACSCalibur
14
Mechanical Pros Cons Cheaper Slow
(?x102cellssec-1) No aerosol 1(2) way
sorting Easier to use Dilute sorted
sample Multiparameter
15
Electrostatic  Mechanical  Lab-on-chip
16
Gawad, S., Heuschkel, M., Leung-Ki, Y.,
Iuzzolino, R., Schild, L., Lerch, Ph. Renaud,
Ph.Fabrication of a microfluidic cell ananlyzer
in a microchannel using impedance spectroscopy in
IEEE-EMBS Conference on Microtechnologies in
Medicine Biology 297-301 (Lyon, France, 2000).
17
(No Transcript)
18
(No Transcript)
19
(No Transcript)
20
Useful texts
Flow cytometry a practical approach MG Ormerod
IRL Press Practical flow cytometry HM Shapiro
WileyLiss Current protocols in flow cytometry Ed
JP Robinson et al Wiley
21
Some cell sorting web resources
Terry Hoy et al Cardiff University
http//www.cardiff.ac.uk/medicine/haematology/cyto
netuk/introduction_to_fcm/cell_sorting.htm
Royal Microscopical Society
http//www.rms.org.uk/cyto.shtml
Derek Davies et al London Research Institute
http//science.cancerresearchuk.org/sci/facs/fac_l
abinfo/flow_sorting/?version1
Cytometry E-mail archive Purdue USA
http//www.cyto.purdue.edu/hmarchiv/index.htm