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      From Animaculum to single molecules: 300 years of the light microscope


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          Although not laying claim to being the inventor of the light microscope, Antonj van Leeuwenhoek (1632–1723) was arguably the first person to bring this new technological wonder of the age properly to the attention of natural scientists interested in the study of living things (people we might now term ‘biologists’). He was a Dutch draper with no formal scientific training. From using magnifying glasses to observe threads in cloth, he went on to develop over 500 simple single lens microscopes (Baker & Leeuwenhoek 1739 Phil. Trans. 41, 503–519. ( doi:10.1098/rstl.1739.0085)) which he used to observe many different biological samples. He communicated his finding to the Royal Society in a series of letters (Leeuwenhoek 1800 The select works of Antony Van Leeuwenhoek, containing his microscopical discoveries in many of the works of nature , vol. 1) including the one republished in this edition of Open Biology. Our review here begins with the work of van Leeuwenhoek before summarizing the key developments over the last ca 300 years, which has seen the light microscope evolve from a simple single lens device of van Leeuwenhoek's day into an instrument capable of observing the dynamics of single biological molecules inside living cells, and to tracking every cell nucleus in the development of whole embryos and plants.

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          Most cited references37

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          Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy.

          Recent advances in far-field fluorescence microscopy have led to substantial improvements in image resolution, achieving a near-molecular resolution of 20 to 30 nanometers in the two lateral dimensions. Three-dimensional (3D) nanoscale-resolution imaging, however, remains a challenge. We demonstrated 3D stochastic optical reconstruction microscopy (STORM) by using optical astigmatism to determine both axial and lateral positions of individual fluorophores with nanometer accuracy. Iterative, stochastic activation of photoswitchable probes enables high-precision 3D localization of each probe, and thus the construction of a 3D image, without scanning the sample. Using this approach, we achieved an image resolution of 20 to 30 nanometers in the lateral dimensions and 50 to 60 nanometers in the axial dimension. This development allowed us to resolve the 3D morphology of nanoscopic cellular structures.
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            Negative refraction makes a perfect lens

            With a conventional lens sharpness of the image is always limited by the wavelength of light. An unconventional alternative to a lens, a slab of negative refractive index material, has the power to focus all Fourier components of a 2D image, even those that do not propagate in a radiative manner. Such "superlenses" can be realized in the microwave band with current technology. Our simulations show that a version of the lens operating at the frequency of visible light can be realized in the form of a thin slab of silver. This optical version resolves objects only a few nanometers across.
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              Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy.

              A long-standing goal of biology is to map the behavior of all cells during vertebrate embryogenesis. We developed digital scanned laser light sheet fluorescence microscopy and recorded nuclei localization and movement in entire wild-type and mutant zebrafish embryos over the first 24 hours of development. Multiview in vivo imaging at 1.5 billion voxels per minute provides "digital embryos," that is, comprehensive databases of cell positions, divisions, and migratory tracks. Our analysis of global cell division patterns reveals a maternally defined initial morphodynamic symmetry break, which identifies the embryonic body axis. We further derive a model of germ layer formation and show that the mesendoderm forms from one-third of the embryo's cells in a single event. Our digital embryos, with 55 million nucleus entries, are provided as a resource.

                Author and article information

                Open Biol
                Open Biol
                Open Biology
                The Royal Society
                April 2015
                29 April 2015
                29 April 2015
                : 5
                : 4
                : 150019
                Biological Physical Sciences Institute (BPSI), Departments of Physics and Biology, University of York , York YO10 5DD, UK
                Author notes

                An invited review to commemorate 350 years of scientific publishing at the Royal Society.

                Author information

                © 2015 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

                : 30 January 2015
                : 1 April 2015
                Review Article
                Custom metadata
                April 2015

                Life sciences
                optical microscopy,superresolution,fluorescence
                Life sciences
                optical microscopy, superresolution, fluorescence


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