Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye. There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy.
A microscope is a laboratory instrument used to examine objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using a microscope. Microscopic means being invisible to the eye unless aided by a 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.
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 within an object. This technique is used extensively in the scientific and industrial communities and typical applications are in life sciences, semiconductor inspection and materials science.
William Esco Moerner, also known as W. E. Moerner, is an American physical chemist and chemical physicist with current work in the biophysics and imaging of single molecules. He is credited with achieving the first optical detection and spectroscopy of a single molecule in condensed phases, along with his postdoc, Lothar Kador. Optical study of single molecules has subsequently become a widely used single-molecule experiment in chemistry, physics and biology. In 2014, he was awarded the Nobel Prize in Chemistry.
Richard Henderson is a British molecular biologist and biophysicist and pioneer in the field of electron microscopy of biological molecules. Henderson shared the Nobel Prize in Chemistry in 2017 with Jacques Dubochet and Joachim Frank."Thanks to his work, we can look at individual atoms of living nature, thanks to cryo-electron microscopes we can see details without destroying samples, and for this he won the Nobel Prize in Chemistry."
Stefan Walter Hell is a Romanian-German physicist and one of the directors of the Max Planck Institute for Multidisciplinary Sciences in Göttingen, and of the Max Planck Institute for Medical Research in Heidelberg, both of which are in Germany. He received the Nobel Prize in Chemistry in 2014 "for the development of super-resolved fluorescence microscopy", together with Eric Betzig and William Moerner.
Janelia Research Campus is a scientific research campus of the Howard Hughes Medical Institute that opened in October 2006. The campus is located in Loudoun County, Virginia, near the town of Ashburn. It is known for its scientific research and modern architecture. The current executive director of the laboratory is Ronald Vale, who is also a vice-president of HHMI. He succeeded Gerald M. Rubin in 2020. The campus was known as "Janelia Farm Research Campus" until 2014.
Christoph Cremer is a German physicist and emeritus at the Ruprecht-Karls-University Heidelberg, former honorary professor at the University of Mainz and was a former group leader at Institute of Molecular Biology (IMB) at the Johannes Gutenberg University of Mainz, Germany, who has successfully overcome the conventional limit of resolution that applies to light based investigations by a range of different methods. In the meantime, according to his own statement, Christoph Cremer is a member of the Max Planck Institute for Chemistry and the Max Planck Institute for Polymer Research.
Jennifer Lippincott-Schwartz is a Senior Group Leader at Howard Hughes Medical Institute's Janelia Research Campus and a founding member of the Neuronal Cell Biology Program at Janelia. Previously, she was the Chief of the Section on Organelle Biology in the Cell Biology and Metabolism Program, in the Division of Intramural Research in the Eunice Kennedy Shriver National Institute of Child Health and Human Development at the National Institutes of Health from 1993 to 2016. Lippincott-Schwartz received her PhD from Johns Hopkins University, and performed post-doctoral training with Richard Klausner at the NICHD, NIH in Bethesda, Maryland.
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 or on the far-field. Among techniques that rely on the latter are those that improve the resolution only modestly beyond the diffraction-limit, such as confocal microscopy with closed pinhole or aided by computational methods such as deconvolution or detector-based pixel reassignment, the 4Pi microscope, and structured-illumination microscopy technologies such as SIM and SMI.
Xiaowei Zhuang is a Chinese-American biophysicist who is the David B. Arnold Jr. Professor of Science, Professor of Chemistry and Chemical Biology, and Professor of Physics at Harvard University, and an Investigator at the Howard Hughes Medical Institute. She is best known for her work in the development of Stochastic Optical Reconstruction Microscopy (STORM), a super-resolution fluorescence microscopy method, and the discoveries of novel cellular structures using STORM. She received a 2019 Breakthrough Prize in Life Sciences for developing super-resolution imaging techniques that get past the diffraction limits of traditional light microscopes, allowing scientists to visualize small structures within living cells. She was elected a Member of the American Philosophical Society in 2019 and was awarded a Vilcek Foundation Prize in Biomedical Science in 2020.
Photo-activated localization microscopy and stochastic optical reconstruction microscopy (STORM) are widefield fluorescence microscopy imaging methods that allow obtaining images with a resolution beyond the diffraction limit. The methods were proposed in 2006 in the wake of a general emergence of optical super-resolution microscopy methods, and were featured as Methods of the Year for 2008 by the Nature Methods journal. The development of PALM as a targeted biophysical imaging method was largely prompted by the discovery of new species and the engineering of mutants of fluorescent proteins displaying a controllable photochromism, such as photo-activatible GFP. However, the concomitant development of STORM, sharing the same fundamental principle, originally made use of paired cyanine dyes. One molecule of the pair, when excited near its absorption maximum, serves to reactivate the other molecule to the fluorescent state.
Lattice light-sheet microscopy is a modified version of light sheet fluorescence microscopy that increases image acquisition speed while decreasing damage to cells caused by phototoxicity. This is achieved by using a structured light sheet to excite fluorescence in successive planes of a specimen, generating a time series of 3D images which can provide information about dynamic biological processes.
Expansion microscopy (ExM) is a sample preparation tool for biological samples that allows investigators to identify small structures by expanding them using a polymer system. The premise is to introduce a polymer network into cellular or tissue samples, and then physically expand that polymer network using chemical reactions to increase the size of the biological structures. Among other benefits, ExM allows those small structures to be imaged with a wider range of microscopy techniques. It was first proposed in a 2015 article by Fei Chen, Paul W. Tillberg, and Edward Boyden. Current research allows for the expansion of samples up to 16x larger than their initial size. This technique has been found useful in various laboratory settings, such as analyzing biological molecules. ExM allows researchers to use standard equipment in identifying small structures, but requires following of procedures in order to ensure clear results.
Michael Wesley Davidson was an American research scientist and microscopist. He used microscopes to create images of crystallized substances like DNA and hormones, and he contributed to Nobel Prize-honored research about the inner workings of cells. He is credited by 2014 Nobel Laureate Eric Betzig with teaching Betzig and fellow researcher Harald Hess about fluorescent proteins and providing the samples that led to the development of photoactivated localization microscopy (PALM), a super-resolution microscopy technique.
Ibrahim I. Cissé is an Nigerien-American biophysicist. He is currently director of the Max Planck Institute of Immunobiology and Epigenetics. Previously, Cissé was at the California Institute of Technology as Professor of Physics and the Massachusetts Institute of Technology as Professor of Physics and Biology. He has won several awards for his work, including in 2021 the MacArthur Fellowship.
Na Ji is an American biophysicist and the Luis Alvarez Memorial Chair in Experimental Physics at UC Berkeley, where her work focuses on optical microscopy techniques for in vivo imaging and biophotonics. She has a joint appointment as faculty scientist at Lawrence Berkeley National Laboratory.
Cryomicroscopy is a technique in which a microscope is equipped in such a fashion that the object intended to be inspected can be cooled to below room temperature. Technically, cryomicroscopy implies compatibility between a cryostat and a microscope. Most cryostats make use of a cryogenic fluid such as liquid helium or liquid nitrogen. There exists two common motivations for performing a cryomicroscopy. One is to improve upon the process of performing a standard microscopy. Cryogenic electron microscopy, for example, enables the studying of proteins with limited radiation damage. In this case, the protein structure may not change with temperature, but the cryogenic environment enables the improvement of the electron microscopy process. Another motivation for performing a cryomicroscopy is to apply the microscopy to a low-temperature phenomenon. A scanning tunnelling microscopy under a cryogenic environment, for example, allows for the studying of superconductivity, which does not exist at room temperature.
Harald Frederick Hess is an American physicist and Senior Group Leader at Howard Hughes Medical Institute's Janelia Research Campus, known for his work in scanning probe microscopy, light microscopy and electron microscopy.
