Lecture

Phase-Contrast Microscopy: Principles and Applications

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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 phase-contrast microscopy, explaining how positive and negative phase contrast work to enhance the visibility of different objects based on their refractive index. It also delves into the implementation and alignment of phase-contrast microscopy, highlighting the importance of correct adjustment. Additionally, the lecture explores polarization microscopy, contrasting different contrast techniques in microscopy and the use of polarized light to reveal molecular orders in various samples. The discussion extends to the generation and manipulation of linearly polarized light, the role of polarizers and analyzers, and the configuration of a polarized light microscope in biology.

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

Ontological neighbourhood

Biology

Anatomy: Histology

Physics

Optics: Wave polarization

Related concepts (30)

Polarization (physics)

Polarization (also polarisation) is a property of transverse waves which specifies the geometrical orientation of the oscillations. In a transverse wave, the direction of the oscillation is perpendicular to the direction of motion of the wave. A simple example of a polarized transverse wave is vibrations traveling along a taut string (see image); for example, in a musical instrument like a guitar string. Depending on how the string is plucked, the vibrations can be in a vertical direction, horizontal direction, or at any angle perpendicular to the string.

Microscopy

Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection, or refraction of electromagnetic radiation/electron beams interacting with the specimen, and the collection of the scattered radiation or another signal in order to create an image.

Phase-contrast imaging

Phase-contrast imaging is a method of that has a range of different applications. It measures differences in the refractive index of different materials to differentiate between structures under analysis. In conventional light microscopy, phase contrast can be employed to distinguish between structures of similar transparency, and to examine crystals on the basis of their double refraction. This has uses in biological, medical and geological science.

Phase-contrast X-ray imaging

Phase-contrast X-ray imaging or phase-sensitive X-ray imaging is a general term for different technical methods that use information concerning changes in the phase of an X-ray beam that passes through an object in order to create its images. Standard X-ray imaging techniques like radiography or computed tomography (CT) rely on a decrease of the X-ray beam's intensity (attenuation) when traversing the sample, which can be measured directly with the assistance of an X-ray detector.

Quantitative phase-contrast microscopy

FORCETOC Quantitative phase contrast microscopy or quantitative phase imaging are the collective names for a group of microscopy methods that quantify the phase shift that occurs when light waves pass through a more optically dense object. Translucent objects, like a living human cell, absorb and scatter small amounts of light. This makes translucent objects much easier to observe in ordinary light microscopes. Such objects do, however, induce a phase shift that can be observed using a phase contrast microscope.

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Polarized Light Microscopy: Techniques and Applications

Explores polarized light microscopy techniques, including observing spherulites and birefringence in specimens, and analyzing various materials.

Optical Microscopy: Concepts and Applications

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