Lecture

Microscopy Image Analysis

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In course

BIO-221: Cell and developmental biology for engineers

Students will learn essentials of cell and developmental biology with an engineering mind set, with an emphasis on animal systems and quantitative approaches. ...

Description

This lecture covers the principles of light microscopy, focusing on the use of fluorescent proteins for cellular imaging. It delves into the concepts of GFP/RFP mutations, applications of fluorescent proteins, optical slicing methods, and modulation transfer functions. The instructor discusses the challenges in imaging thick specimens, the benefits of timelapse microscopy, and the techniques for signal and sampling in microscopy. Moreover, the lecture explores the advancements in spinning disk confocal microscopy and the resolution limits in light microscopy.

Instructors (2)

Pierre Gönczy

Pierre Gönczy obtained his PhD from The Rockefeller University (New York, USA) in 1995 and joined the laboratory of Tony Hyman at the EMBL (Heidelberg, Germany) as a postdoctoral fellow in 1996. He started his own laboratory at ISREC in 2000. In 2005, he became Associate Professor at the EPFL School of Life Sciences, and was promoted Full Professor in 2009.

Andrew Charles Oates

After an undergraduate degree in Biochemistry at the University of Adelaide with Honours in Robert Saint’s lab, Andrew Oates received his Ph.D. at the Ludwig Institute for Cancer Research and the University of Melbourne in the lab of Andrew Wilks. His postdoctoral time was at Princeton University and the University of Chicago in the lab of Robert Ho, where his studies on the segmentation clock in zebrafish began in 1998. In 2003 he moved to Germany and started his group at the Max Planck Institute for Molecular Cell Biology and Genetics in Dresden. In 2012 he accepted a position at University College London as Professor of vertebrate developmental genetics and moved his group to the MRC-National Institute for Medical Research at Mill Hill in London. From April 2015, he became a member of the Francis Crick Institute in London. In September 2016, he joined École polytechnique fédéral de Lausanne (EPFL) in Switzerland as a Professor, where he is the head of the Timing, Oscillation, Patterns Laboratory. From April 2018 he served as Director of the Institute of Bioengineering, and from January 2021 became the Dean of the School of Life Sciences. The Timing, Oscillation, Patterns Laboratory is composed of biologists, engineers, and physicists using molecular genetics, quantitative imaging, and theoretical analysis to study a population of coupled genetic oscillators in the vertebrate embryo termed the segmentation clock. This system drives the rhythmic, sequential, and precise formation of embryonic body segments, exhibiting rich spatial and temporal phenomena spanning from molecular to tissue scales.

Official source

Ontological neighbourhood

Biology

Anatomy: Histology

Related concepts (32)

Green fluorescent protein

The green fluorescent protein (GFP) is a protein that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP. However, GFPs have been found in other organisms including corals, sea anemones, zoanithids, copepods and lancelets. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm.

Confocal microscopy

Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object.

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.

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.

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.

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