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Lecture

Microscopy: Fluorescence & Resolution

In course

MICRO-310(b): Signals and systems I (for SV)

Présentation des concepts et des outils de base pour l'analyse et la caractérisation des signaux, la conception de systèmes de traitement et la modélisation linéaire de systèmes pour les étudiants en sciences de la vie. Application de ces techniques au tra ...

Description

This lecture covers the principles of microscopy, focusing on fluorescence imaging and resolution. It discusses the use of fluorophores and genetic methods for specific molecular probes, as well as the components of a fluorescence microscope. The lecture delves into the physics of light propagation, diffraction patterns, and the concept of impulse response in microscopy. It also explores the frequency response and resolution criteria in microscopy, including the Rayleigh criterion. The presentation concludes with a mention of the Nobel Prize in Chemistry awarded for achievements in super-resolved fluorescence microscopy.

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Instructor

Michaël Unser

Official source

Related concepts (32)

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.

Diffraction

Diffraction is the interference or bending of waves around the corners of an obstacle or through an aperture into the region of geometrical shadow of the obstacle/aperture. The diffracting object or aperture effectively becomes a secondary source of the propagating wave. Italian scientist Francesco Maria Grimaldi coined the word diffraction and was the first to record accurate observations of the phenomenon in 1660.

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.

Fraunhofer diffraction

In optics, the Fraunhofer diffraction equation is used to model the diffraction of waves when plane waves are incident on a diffracting object, and the diffraction pattern is viewed at a sufficiently long distance (a distance satisfying Fraunhofer condition) from the object (in the far-field region), and also when it is viewed at the focal plane of an imaging lens. In contrast, the diffraction pattern created near the diffracting object and (in the near field region) is given by the Fresnel diffraction equation.

Fresnel diffraction

In optics, the Fresnel diffraction equation for near-field diffraction is an approximation of the Kirchhoff–Fresnel diffraction that can be applied to the propagation of waves in the near field. It is used to calculate the diffraction pattern created by waves passing through an aperture or around an object, when viewed from relatively close to the object. In contrast the diffraction pattern in the far field region is given by the Fraunhofer diffraction equation. The near field can be specified by the Fresnel number, F, of the optical arrangement.

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