Mediaspace scheduled maintenance: Aug 25, 2026 07:00 - 12:00 AM. During this time, videos will be temporarily unavailable. Check status updates.

Lecture

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

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.

This video is available exclusively on Mediaspace for a restricted audience. Please log in to MediaSpace to access it if you have the necessary permissions.

Watch on Mediaspace

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 (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.

Phase-contrast microscopy

NOTOC Phase-contrast microscopy (PCM) is an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations. When light waves travel through a medium other than a vacuum, interaction with the medium causes the wave amplitude and phase to change in a manner dependent on properties of the medium.

Super-resolution microscopy

Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that use the Pendry Superlens and near field scanning optical microscopy) or on the far-field.

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.

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.

Related lectures (53)

Transmission Electron Microscopy: Advanced Techniques MOOC: Transmission Electron Microscopy for Materials Sciences

Explores advanced techniques in transmission electron microscopy, focusing on phase contrast imaging and camera settings.

Microscopy Image Analysis

Explores light microscopy principles, fluorescent proteins, optical slicing, and modulation transfer functions in cellular imaging.

Micro and Nanofabrication: Inspection and Metrology MOOC: Micro and Nanofabrication (MEMS)

Explores optical microscopy variations for inspection and dimension measurement in Micro and Nanofabrication processes.

Dark-field Microscopy: Techniques and Applications

Explores dark-field microscopy, highlighting its improved contrast, high sensitivity, and potential for high-resolution imaging in various scientific fields.