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TIRF Microscope (Shared Equipment)
Read the Standard Operating Procedure (SOP) for our TIRF Scope
TIRF (Total Internal Reflection Fluorescence) Microscopy is a very sensitive technique to visualize single
molecules near the surface of a coverglass. Incident laser light is adjusted until reaching the "critical angle" at
which no light is refracted (all incident light is reflected). Despite no direct
illumniation of the sample with the laser light, a so-called "evanescent wave" exists 100 - 200 nm above the
coverslip and provides an excitation source for the sample of interest (with almost no background fluorescence).
We have a Nikon Laser TIRF setup that uses two lasers (488 nm and
543 nm) to detect labeled molecules. The TIRF lasers are mounted
on a Nikon TE2000U Microscope that also allows phase-contrast and epifluorescence techniques to be combined with
TIRF technology.
Our TIRF Scope is a Shared Resource. To gain access to the TIRF Scope:
1. Read the Nikon Introduction to TIRF Microscopy.
Make sure that TIRF is right for your application! Our setup requires samples on coverglass (due to the working distance of the TIRF lenses), and
the excitation working distance is very close (100 - 200 nm) above the surface of the coverslip.
2. Print and read the Standard Operating Procedure for our TIRF scope. Bring this SOP to TIRF
training.
3. Then, contact a graduate student member of the Biomembrane Lab to schedule a time for training/operating the TIRF scope:
Dan Estes - danestes@umich.edu
Jeff Uram - uramj@umich.edu
Sheereen Majd - sheereen@umich.edu
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