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Lecture

Optical Filters: Understanding Properties and Selection

In course

MICRO-561: Fundamentals of biomicroscopy

Introduction to the basic operational principles of the most commonly used optical microscopes such as widefield and fluorescence.
Introduction to essential microscopy components such as lenses, objectives, filters, cameras and light sources. ...

Description

This lecture covers the importance of understanding the properties of optical filters for fluorescence microscopy, including the characterization of transmission/reflection spectra and selecting suitable filter combinations. Examples are provided on choosing filter sets based on absorption and emission spectra, as well as the arrangement of filters in a fluorescence filter cube. The lecture also discusses multi-color imaging using different dyes and the use of filter sets to examine each dye separately.

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Instructors (2)

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

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)

Official source

Ontological neighbourhood

Chemistry

Physical chemistry: Spectroscopy

Related concepts (32)

Optical filter

An optical filter is a device that selectively transmits light of different wavelengths, usually implemented as a glass plane or plastic device in the optical path, which are either dyed in the bulk or have interference coatings. The optical properties of filters are completely described by their frequency response, which specifies how the magnitude and phase of each frequency component of an incoming signal is modified by the filter. Filters mostly belong to one of two categories.

Emission spectrum

The emission spectrum of a chemical element or chemical compound is the spectrum of frequencies of electromagnetic radiation emitted due to an electron making a transition from a high energy state to a lower energy state. The photon energy of the emitted photon is equal to the energy difference between the two states. There are many possible electron transitions for each atom, and each transition has a specific energy difference. This collection of different transitions, leading to different radiated wavelengths, make up an emission spectrum.

Absorption spectroscopy

Absorption spectroscopy refers to spectroscopic techniques that measure the absorption of electromagnetic radiation, as a function of frequency or wavelength, due to its interaction with a sample. The sample absorbs energy, i.e., photons, from the radiating field. The intensity of the absorption varies as a function of frequency, and this variation is the absorption spectrum. Absorption spectroscopy is performed across the electromagnetic spectrum.

Fluorophore

A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. Fluorophores typically contain several combined aromatic groups, or planar or cyclic molecules with several π bonds. Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for staining of certain structures, as a substrate of enzymes, or as a probe or indicator (when its fluorescence is affected by environmental aspects such as polarity or ions).

Fluorescence microscope

A fluorescence microscope is an optical microscope that uses fluorescence instead of, or in addition to, scattering, reflection, and attenuation or absorption, to study the properties of organic or inorganic substances. "Fluorescence microscope" refers to any microscope that uses fluorescence to generate an image, whether it is a simple set up like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image.

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