Journal
JOURNAL OF CELL SCIENCE
Volume 123, Issue 21, Pages 3621-3628Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.056218
Keywords
Total internal reflection fluorescence microscopy; Evanescent wave microscopy; Evanescent field microscopy; Fluorescence
Categories
Funding
- BBSRC [BB/H002308/1]
- Advantage West Midlands (AWM)
- NIH [R01 GM087977]
- BBSRC [BB/H002308/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/H002308/1] Funding Source: researchfish
Ask authors/readers for more resources
Total internal reflection fluorescence (TIRF) microscopy can be used in a wide range of cell biological applications, and is particularly well suited to analysis of the localization and dynamics of molecules and events near the plasma membrane. The TIRF excitation field decreases exponentially with distance from the cover slip on which cells are grown. This means that fluorophores close to the cover slip (e.g. within similar to 100 nm) are selectively illuminated, highlighting events that occur within this region. The advantages of using TIRF include the ability to obtain high-contrast images of fluorophores near the plasma membrane, very low background from the bulk of the cell, reduced cellular photodamage and rapid exposure times. In this Commentary, we discuss the applications of TIRF to the study of cell biology, the physical basis of TIRF, experimental setup and troubleshooting.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available