Fluorescence and the World it has Opened
Reading Assignments:
The Molecular Probes Handbook Fluorescence Fundamentals
Fluorescence microscopy – Lichtman & Conchello.pdf (or http://www.nature.com/nmeth/journal/v2/n12/full/nmeth817.html)
Jablonski Diagram
Reading assignment: Jablonski Diagram.pdf
(or https://chem.libretexts.org/Core/Physical_and_Theoretical_Chemistry/Spectroscopy/Electronic_Spectroscopy/Jablonski_diagram)
Ground state
Excitation
Excited state
- Collisional quenching
- Fluorescence resonance energy transfer
- Fluorescence excited-state lifetime
Inter system crossing to triplet state
- Photobleaching
- Photo activation
Emission
Molar extinction coefficient (e) or EC
Quantum Yield (QY)
Fluorescence intensity per dye molecule is proportional to EC X QY
Stokes Shift
Excitation Spectrum
Emission Spectrum
Fluorescence Excitation Sources
Reading Assignments:
Light Sources.pdf
(or http://zeiss-campus.magnet.fsu.edu/articles/lightsources/lightsourcefundamentals.html)
Semrock – Optical Filters.pdf
(or http://www.semrock.com/1optical-filters-for-laser-based-fluorescence-microscopes.aspx)
Table 1: Lasers for popular fluorophores. “DPSS” = diode-pumped solid-state laser. “OPS” = optically pumped semiconductor laser.
“Doubled” = frequency doubled via a nonlinear optical crystal.
Fluorescence Filters
Reading assignment:
Optical Filters Handbook – Reichman.pdf (or http://www.aic-imagecentral.com/products/pdfs/hdbk4.pdf)
Fluorescence Detectors
http://en.wikipedia.org/wiki/Photomultiplier
Hamamatsu PMT handbook.pdf
(or http://sales.hamamatsu.com/assets/applications/ETD/pmt_handbook/pmt_handbook_complete.pdf)
Hamamatsu Cameras.pdf (or http://www.emez.ethz.ch/education/pastevents/pastev/Cameras_rf.pdf)
Fluorescent Materials in Biology
Reading assignment:
The Fluorescent Toolbox.pdf (“The Fluorescent Toolbox” PMID: 16614209)
Fluorescent dyes
Fluorescent Proteins
Quantum Dots
Fluorescent Antibodies