Lighting up the cell surface with evanescent wave microscopy

Area of Science:

• Cell Biology
• Microscopy Techniques
• Biophysics

Background:

• Evanescent wave microscopy, also known as total internal reflection fluorescence microscopy (TIR-FM), is crucial for studying cellular events at the plasma membrane.
• It allows direct observation of dynamic processes like synaptic vesicle fusion and single-molecule tracking during signal transduction.
• The technique's popularity has grown due to advancements like green-fluorescent protein (GFP) and improved instrumentation.

Purpose of the Study:

• This review focuses on recent technical developments in TIR-FM.
• It also highlights the latest applications of this powerful microscopy technique.

Main Methods:

• The review discusses advancements in evanescent wave microscopy instrumentation.
• It covers the use of fluorescent markers, particularly green-fluorescent protein (GFP).
• Applications reviewed include observing membrane dynamics and molecular events near the cell surface.

Main Results:

• TIR-FM enables high-resolution visualization of cellular processes occurring in the immediate vicinity of the plasma membrane.
• The integration of GFP has significantly enhanced the capabilities and accessibility of TIR-FM.
• New technical developments have expanded the scope of biological questions addressable by TIR-FM.

Conclusions:

• Evanescent wave microscopy is a vital tool for understanding membrane-proximal cellular functions.
• Continued technical innovation is driving new applications and insights in cell biology.
• TIR-FM, especially with GFP, offers unprecedented views into dynamic cellular events.