Lecture

Total Internal Reflection Fluorescence Microscopy

Graph Chat

Chat with Graph Search

Ask any question about EPFL courses, lectures, exercises, research, news, etc. or try the example questions below.

DISCLAIMER: The Graph Chatbot is not programmed to provide explicit or categorical answers to your questions. Rather, it transforms your questions into API requests that are distributed across the various IT services officially administered by EPFL. Its purpose is solely to collect and recommend relevant references to content that you can explore to help you answer your questions.

In course

MICRO-562: Experimental biomicroscopy

By combining hands-on and theoretical training, this course introduces commonly used optical microscopy techniques. The lab training takes place in BIOP Core Facility and provides access to use microscopes and image biological samples. Theoretical part cov ...

Description

This lecture covers the principles and applications of Total Internal Reflection Fluorescence Microscopy (TIRFM). It explains how TIRFM uses evanescent waves to selectively excite fluorophores near the coverslip surface, providing high-contrast images. The lecture discusses the limitations of conventional fluorescence microscopy and the conditions required for Total Internal Reflection (TIR). It also explores the concept of evanescent waves in TIR, the penetration depth of the evanescent wave, and various fluorescence microscopy techniques. Furthermore, it delves into the details of Fluorescence Recovery After Photobleaching (FRAP) method, its applications, kinetic analysis, and equipment. The lecture concludes with examples of FRAP experiments and the measurement of diffusion coefficients using FRAP.

Instructors (2)

Arne Seitz

04/1996-06/2000 Scientific co-worker at Philipps-University of Marburg, Dep. of Physical Chemistry 07/2000-10/2002 Post Doc at Max-Planck Unit for Structural Molecular Biology in Hamburg 11/2002-11/2005 Post Doc at European Molecular Biolohy Laboratory (EMBL), Cell Biology and Cell Biophysics Programme 11/2005-03/2009 Staff Scientist at Advanced Light Microscopy Facility, EMBL 04/2009- Head of Bioimaging and Optics platforme (BIOP), Ecole Polytechnique Fédérale de Lausanne (EPFL)

Hatice Altug

2020-current Full Professor at the Institute of Bioengineering, EPFL, Switzerland2013-2020 Associate Professor (with tenure) at the Institute of Bioengineering, EPFL, Switzerland 2013 Associate Professor (with tenure) at Electrical and Computer Engineering Department of Boston University, USA 2007-2013 Assistant Professor (tenure-track) at Electrical and Computer Engineering Department of Boston University, USA 2007 Post-doctoral Fellow at Center for Engineering in Medicine of Harvard Medical School, USA 2000-2007 PhD. in Applied Physics at Stanford University, USA 1996-2000 B.S. in Physics at Bilkent University, Turkey

Official source

Related concepts (26)

Total internal reflection fluorescence microscope

A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide. The technique is based on the principle that when excitation light is totally internally reflected in a transparent solid coverglass at its interface with a liquid medium, an electromagnetic field, also known as an evanescent wave, is generated at the solid-liquid interface with the same frequency as the excitation light.

Total internal reflection

In physics, total internal reflection (TIR) is the phenomenon in which waves arriving at the interface (boundary) from one medium to another (e.g., from water to air) are not refracted into the second ("external") medium, but completely reflected back into the first ("internal") medium. It occurs when the second medium has a higher wave speed (i.e., lower refractive index) than the first, and the waves are incident at a sufficiently oblique angle on the interface.

Confocal microscopy

Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object.

Specular reflection

Specular reflection, or regular reflection, is the mirror-like reflection of waves, such as light, from a surface. The law of reflection states that a reflected ray of light emerges from the reflecting surface at the same angle to the surface normal as the incident ray, but on the opposing side of the surface normal in the plane formed by the incident and reflected rays. This behavior was first described by Hero of Alexandria (AD c. 10–70). Later, Alhazen gave a complete statement of the law of reflection.

Reflection (physics)

Reflection is the change in direction of a wavefront at an interface between two different media so that the wavefront returns into the medium from which it originated. Common examples include the reflection of light, sound and water waves. The law of reflection says that for specular reflection (for example at a mirror) the angle at which the wave is incident on the surface equals the angle at which it is reflected. In acoustics, reflection causes echoes and is used in sonar. In geology, it is important in the study of seismic waves.

Related lectures (49)

Electromagnetic Wave Propagation: Reflection, Refraction, Absorption

Explores electromagnetic wave propagation in matter, focusing on absorption, reflection, and refraction phenomena.

Single Molecule FRET

Introduces Single Molecule FRET, explaining energy transfer between molecules and its applications in studying proteins and nucleic acids.

Optical Microscopy: Concepts and Applications

Explores optical microscopy principles, techniques, resolution limits, and Nobel Prize-winning advancements in microscopy.

Optical Waves: Reflection, Refraction, and Transmission

Explores optical wave behavior, including reflection, refraction, transmission, and intensity effects at interfaces.

Interaction of E-M Waves with Matter: Transmission, Absorption, Reflection