Don't miss these leading Invited Review Papers
Cryo-EM - the first thirty years
J. Dubochet
Thirty years ago, in December 1981, The Journal of Microscopy published a very short paper entitled ‘Vitrification of pure water for electron microscopy’. It turned out to be important for the development of cryo-electron microscopy and it contributed to reverse, from foe to friend, the status of water in electron microscopists’ minds. This change has brought obvious gains. The future will tell how many more are still to come.
Resolution and optical sectioning in the confocal microscope
T.Wilson
In this short review, we present a self-contained discussion of the image formation properties of the fluorescent confocal microscope. The optical sectioning or depth discrimination property is discussed in detail and new analytic formulae are presented, which relate the optical sectioning strength to the wavelength, numerical aperture and pinhole aperture size in a particularly simple fashion.
A simple introduction to multiphoton microscopy
A. Ustione, D.W. Piston
Multiphoton microscopy is a powerful technique based on complex quantum mechanical effects. Thanks to the development of turnkey mode-locked laser systems, multiphoton microscopy is now available for everyone to use without extreme complexity. In this short introduction, we describe qualitatively the important concepts underlying the most commonly used type of multiphoton microscopy (two-photon excitation). We elucidate how those properties lead to the powerful results that have been achieved using this technique. As with any technique, two-photon excitation microscopy has limitations that we describe, and we provide examples of particular classes of experiments where two-photon excitation microscopy is advantageous over other approaches. Finally, we briefly describe other useful multiphoton microscopy approaches, such as three-photon excitation and second harmonic generation imaging.
Quantitative micro_CT
K.Schladitz
In this paper, the field of quantitative microcomputed tomography arising from the combination of microcomputed tomography and quantitative 3D image analysis, is summarized with focus on materials science applications. Opportunities and limitations as well as typical application scenarios are discussed. Selected examples provide an insight into commonly used as well as recent methods from mathematical morphology and stochastic geometry.
Highlights of the optical highlighter fluorescent proteins
G. H. Patterson
The development of super-resolution microscopy techniques using molecular localization, such as photoactivated localization microscopy, fluorescence photoactivated localization microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy with independent running acquisition and many others, has heightened interest in molecules that will be grouped here into a category referred to as ‘optical highlighter’ fluorescent proteins. This review will survey many of the advances in development of fluorescent proteins for optically highlighting sub-populations of fluorescently labelled molecules.
Cryo-electron tomography: methodology, developments and biological applications
D. Vanhecke, S. Asano, Z. Kockovski, R. Fernandez-Busnadiego, N. Schrod, W. Baumeister, V. Lučić
Cryo-electron tomography allows three-dimensional visualization of frozen-hydrated, vitrified biological material at molecular resolution. Here, we summarize the most important sample preparation methods and technical aspects relevant for cryo-electron tomography, as well as its recent biological applications from isolated macromolecular complexes to entire cells and tissues.
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