Lecture

Illumination in Microscopy

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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 introduces the concept of illumination in microscopy, covering critical illumination, Kohler illumination, conjugated planes, and the infinity corrected compound microscope. It explains the importance of field or image forming conjugate planes, the role of aperture in illuminating conjugate planes, and the differences between critical and Kohler illumination methods. The instructor discusses the historical background of August Koehler's illumination system and its impact on image quality. The lecture also addresses the limitations of conventional compound microscopes and the advantages of infinity corrected compound microscopes in producing parallel light wavefronts. Furthermore, it explores the significance of conjugate planes in the context of infinity corrected microscopes.

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

Physics

Optics: Geometrical optics

Biology

Anatomy: Histology

Related concepts (32)

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.

Diaphragm (optics)

In optics, a diaphragm is a thin opaque structure with an opening (aperture) at its center. The role of the diaphragm is to stop the passage of light, except for the light passing through the aperture. Thus it is also called a stop (an aperture stop, if it limits the brightness of light reaching the focal plane, or a field stop or flare stop for other uses of diaphragms in lenses). The diaphragm is placed in the light path of a lens or objective, and the size of the aperture regulates the amount of light that passes through the lens.

Aperture

In optics, an aperture is a hole or an opening through which light travels. More specifically, the aperture and focal length of an optical system determine the cone angle of the bundle of rays that come to a focus in the . An optical system typically has many openings or structures that limit the ray bundles (ray bundles are also known as pencils of light). These structures may be the edge of a lens or mirror, or a ring or other fixture that holds an optical element in place, or may be a special element such as a diaphragm placed in the optical path to limit the light admitted by the system.

Bright-field microscopy

Bright-field microscopy (BF) is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) white light, and contrast in the sample is caused by attenuation of the transmitted light in dense areas of the sample. Bright-field microscopy is the simplest of a range of techniques used for illumination of samples in light microscopes, and its simplicity makes it a popular technique.

Shutter (photography)

In photography, a shutter is a device that allows light to pass for a determined period, exposing photographic film or a photosensitive to light in order to capture a permanent image of a scene. A shutter can also be used to allow pulses of light to pass outwards, as seen in a movie projector or a signal lamp. A shutter of variable speed is used to control exposure time of the film. The shutter is constructed so that it automatically closes after a certain required time interval.

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