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1
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2
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- Lectures: The class consists primarily of lectures and lecture
discussions with mini tutorials as necessary.
- Practicals: There are no practical components to the 631 course. We will
however, look at some instruments and instrument components to gain some
perspectives.
- Quizes: There is a midterm quiz and an end of term quiz. They are worth
60% of the grade.
- Seminar: Each student must
present a seminar at the conclusion of the course. This seminar must be
discussed with the course director prior to preparation. This is worth
30% of the final grade.
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3
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- Flow Cytometry and Sorting, 2nd ed. (M.R. Melamed, T. Lindmo, M.L.
Mendelsohn, eds.), Wiley-Liss, New York, 1990 - referred to here as MLM
- Flow Cytometry: Instrumentation and Data Analysis (M.A. Van Dilla, P.N.
Dean, O.D. Laerum, M.R. Melamed, eds.), Academic Press, London, 1985 –
referred to as VDLM
- Practical Flow Cytometry 3nd edition (1994),H. Shapiro: Alan R. Liss,
New York - referred to as PFC
- Introduction to Flow Cytometry. J. Watson, Cambridge Press, 1991
referred to as IFC
- Methods in Cell Biology: v.40,41, 63, 64 Darzynkiewicz, Robinson &
Crissman, Academic Press, 1994, 2000 MCB
- Data Analysis in Flow Cytometry:A Dynamic Approach-Book on CDROM M. Ormerod referred to as DAFC
- Flow Cytometry: First Principles. (2nd Ed) Alice Longobardi
Givan, Wiley-Liss, 2001 referred to as AFCFP
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4
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- Flow Cytometry and Sorting, 2nd ed. (M.R. Melamed, T. Lindmo, M.L.
Mendelsohn, eds.), Wiley-Liss, New York, 1990 - referred to here as MLM
- Flow Cytometry: Instrumentation and Data Analysis (M.A. Van Dilla, P.N.
Dean, O.D. Laerum, M.R. Melamed, eds.), Academic Press, London, 1985 - VDLM
- Practical Flow Cytometry 4th edition (2003),H. Shapiro: Alan R. Liss,
New York - PFC
- Introduction to Flow Cytometry. J. Watson, Cambridge Press, 1991 IFC
- Methods in Cell Biology: v.40,41, 62, 63 Darzynkiewicz, Robinson &
Crissman, Academic Press, 1994,
(Vol 62,63, 2000) MCB
- Data Analysis in Flow Cytometry:A Dynamic Approach-Book on CDROM M. Ormerod DAFC
- Flow Cytometry: First Principles. 2nd Edition Alice
Longobardi Givan, Wiley-Liss, 2000 AFCFP
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- For help with protocols there are several choices including the MCB
references on the previous slide (Methods in Cell Biology)
- The Handbook of Flow Cytometry Methods
- Current Protocols in Cytometry
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6
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7
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- Powerpoint presentations references as J.Paul Robinson (JPR); Robert
Murphry (RFM), Carleton Stewart (CS)
- Web sources of these presentation are:
- http://www.cyto.purdue.edu/flowcyt/educate/pptslide.htm
- http://www.cyto.purdue.edu/flowcyt/educate1.htm
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9
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10
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11
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12
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- Detection Systems
- Photomultiplier Tubes (PMTs)
- Historically 1-2
- Current Instruments 3-9
- Diodes
- Illumination Systems
- Lasers (350-363, 405, 420, 457, 488, 514, 532, 600, 633 nm)
- Argon ion, Krypton ion, HeNe, HeCd, Yag
- Arc Lamps
- Mercury, Mercury-Xenon (most lines)
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14
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15
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- Coons et al 1941 - developed the fluorescence antibody technique - they labeled antipneumococcal
antibodies with anthracine allowing them to detect both the organism and
the antibody in tissue using UV excited blue fluorescence
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- (1944) Oswald T. Avery
(1887-1955) - demonstrated that DNA was the carrier of genetic
information
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- Developed a flow cytometer for detection of bacteria in aerosols
- Published paper in 1947 (work was done during WWII and was classified).
- Goal was rapid identification of airborne bacteria and spores used in
biological warfare
- Instrument: Sheath of filtered air flowing through a dark-field flow
illuminated chamber. Light source
was a Ford headlamp, PMT detector (very early use of PMT)
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- Friedman (1950) - combined acid fuchsin, acridine yellow and berberine
for uterine cancer detection
using fluorescence microscopy
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25
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26
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27
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28
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- Introduction to course
- Reading and Support Materials
- History
- Technical Highlights
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Notes
