Lecture

Microscopy Resolution: Understanding Bright-Field and Dark-Field Techniques

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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 fundamental concepts of microscopy resolution, focusing on the difference between ideal and real image formation, the impact of aperture on resolution, and the role of numerical aperture in enhancing resolution. It also delves into the principles of bright-field microscopy, emphasizing the importance of contrast and resolution trade-offs. Furthermore, it explores dark-field microscopy as a technique to enhance contrast for transparent specimens, detailing the setup and advantages of this method. The lecture concludes with a discussion on Rheinberg illumination, a specialized dark-field variant that uses colored stops to create striking color contrast in microscopy.

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

Related concepts (31)

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.

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.

F-number

An f-number is a measure of the light-gathering ability of any optical system like a camera lens or even the human eye. It is calculated by dividing the system's focal length by the diameter of the entrance pupil. The f-number is also known as the focal ratio, f-ratio, or f-stop, and it is key in determining the depth of field, rate of light scattering, and exposure of a photograph. The f-number is dimensionless that is usually expressed using a lower-case hooked f with the format N, where N is the f-number.

Optical microscope

The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast. The object is placed on a stage and may be directly viewed through one or two eyepieces on the microscope.

Synthetic-aperture radar

Synthetic-aperture radar (SAR) is a form of radar that is used to create two-dimensional images or three-dimensional reconstructions of objects, such as landscapes. SAR uses the motion of the radar antenna over a target region to provide finer spatial resolution than conventional stationary beam-scanning radars. SAR is typically mounted on a moving platform, such as an aircraft or spacecraft, and has its origins in an advanced form of side looking airborne radar (SLAR).

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