Resources

Fluorescence Microscopy & Imaging Center

This section* is designed to help you gain a good conceptual understanding of the many techniques that are supported in the NIEHS Fluorescence Microscopy and Imaging Center. Below is a list of Web links that you can explore to learn more about many of these techniques. It is suggested that you familiarize yourself with the first three in this list before delving into the more advanced topics. If you are already a FMIC user, you might be able to find more technical information on the confocal file server.

Fluorescence

• Wikipedia, the free encyclopedia: Fluorescence(http://en.wikipedia.org/wiki/Fluorescence)
• Molecular Expressions Microscopy Primer: Introduction to Fluorescence(http://micro.magnet.fsu.edu/primer/lightandcolor/fluorointroduction.html)
• Invitrogen (Molecular Probes): Introduction to Fluorescence(http://probes.invitrogen.com/resources/education/tutorials/1Introduction/player.html)
• J. Paul Robinson’s lecture slides (Purdue University): Fluorescence and Fluorescence Probes(http://www.cyto.purdue.edu/flowcyt/educate/confocal/524Lec3/intro.htm)

Conventional fluorescence microscopy

• Botany Online: Fluorescence Microscopy(http://www.biologie.uni-hamburg.de/b-online/e03/03b.htm)
• C&L Instruments, Inc.: Tutorial – Fluorescence Microscope Optics(http://www.fluorescence.com/tutorial/fm-optic.htm)
• Howstuffworks: How Light Microscopes Work(http://www.howstuffworks.com/light-microscope4.htm)
• Confocal microscopy(http://www.niehs.nih.gov/research/atniehs/core/fmic/mcroscopy.cfm)

Multi-photon (2-photon) excitation microscopy

• Laboratory for Optical and Computational Instrumentation(http://www.loci.wisc.edu/multiphoton/mp.html)
• DRBIO Web Research Group(http://www.drbio.cornell.edu/MPE/mpe1.html)
• Olympus Microscopy Resource Center(http://www.olympusmicro.com/primer/java/multiphoton/jablonski/index.html)
• Olympus Microscopy Resource Center: Specialized Techniques(http://www.olympusmicro.com/primer/java/multiphoton/excitationbleaching/index.html)
• Wiley InterScience: Multi-Photon Imaging(http://www.mrw.interscience.wiley.com/cp/cpcy/articles/cy0209/frame.html)

DIC (Differential Interference Contrast, also called Nomarski) microscopy

• Nikon Microscopy: Phase Contrast and DIC Comparison Image Gallery(http://www.microscopyu.com/galleries/dicphasecontrast/index.html)
• Nikon Microscopy U: Comparison of Phase Contrast and DIC Microscopy(http://www.microscopyu.com/tutorials/java/phasedicmorph/index.html)
• Molecular Expressions Microscopy Primer(http://microscope.fsu.edu/primer/techniques/dic/dicintro.html)

Quantitative colocalization image analysis

• Apple: Colocalization in fluorescence microscopy(http://homepage.mac.com/colocalizerpro/colocalization_b.html)
• Olympus: Colocalization of Fluorophores in Confocal Microscopy(http://www.olympusfluoview.com/applications/colocalization.html)

3D deconvolution of microscope images

• Scientific Volume Imaging B.V. The Knowledge Database(http://support.svi.nl/wiki/DoingDeconvolution)

Localized photolysis and photoactivation

• Brittanica Nobel Prizes: Photolysis(http://www.britannica.com/nobel/micro/465_63.html)
• Ion Channels: Flash photolysis of caged compounds: new tools for cellular physiology(http://www.ionchannels.org/showabstract.php?pmid=2469211)
• National Institute for Medical Research: Caged Compounds(http://www.nimr.mrc.ac.uk/physbiochem/corrie/caged/)
• Invitrogen(http://probes.invitrogen.com/handbook/sections/0503.html)

FRAP (Fluorescence Recovery After Photobleaching)

• BBSRC BioImaging Facility for F.R.A.P. Studies(http://srs.dl.ac.uk/VUV/home-page/BIOIMAGING/Principle%20of%20FRAP.htm)
• Ion Channel Literature(http://www.ionchannels.org/showabstract.php?pmid=2469211)
• Davidson College: Fluorescence Recovery After Photobleaching (FRAP)(http://www.bio.davidson.edu/Courses/Molbio/FRAPx/FRAP.html)

FRET (Förster Resonance Energy Transfer)

• KECK Center for Cellular Imaging(http://www.kcci.virginia.edu/FRET-FLIM/index.php)
• Invitrogen: Technical Focus: Fluorescence Resonance Energy Transfer (FRET)(http://probes.invitrogen.com/handbook/boxes/0422.html)
• University of Delaware: Department of Biological Sciences(http://www.udel.edu/bio/research/facilities/microscopy/tutorials/fret/)
• Molecular Expressions: Fluorescence Resonance Energy Transfer (FRET) Microscopy(http://micro.magnet.fsu.edu/primer/techniques/fluorescence/fret/fretintro.html)
• InterScience: Principles of Resonance Energy Transfer(http://www.mrw.interscience.wiley.com/cp/cpcy/articles/cy0112/frame.html)

FLIM (Fluorescence Lifetime Imaging)

• Trinity College: Fluorescence Lifetime Imaging(http://www.tcd.ie/Biochemistry/confocal/applications.php)
• Olympus: Fluorescence Lifetime Imaging Microscopy (FLIM)(http://www.olympusfluoview.com/applications/flimintro.html)
• High Performance Photon Counting(http://www.becker-hickl.de/applications.htm#Environment)

TIRF (Total Internal Reflection-induced Fluorescence)

• Northwestern University Cell Imaging Facility(http://www.feinberg.northwestern.edu/cif/tirf.htm)
• Nikon Microscopy U: TIRF Microscopy: Introduction and Applications(http://www.microscopyu.com/articles/fluorescence/tirf/tirfintro.html)
• Molecular Expressions(http://micro.magnet.fsu.edu/primer/techniques/fluorescence/tirf/tirfintro.html)

FAIM (Fluorescence Anisotropy Imaging)

• Fluorescence Anisotropy(http://www.urmc.rochester.edu/smd/Rad/foster/research/fluorescenceAnisotropy/index.html)

* The inclusion of Web sites by vendors does not indicate any endorsement of those vendors' products. Rather, these are just the best Web sites the center has found, after a brief Web search, to explain the concepts. Please contact the center if you have found better Web sites.

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