Lipids and the membranes they form are fundamental building blocks of cellular life, and their geometry and chemical properties distinguish membranes from other cellular environments. Collective processes occurring within membranes strongly impact cellular behavior and biochemistry, and understanding these processes presents unique challenges due to the often complex and myriad interactions between membrane components. Super-resolution microscopy offers a significant gain in resolution over traditional optical microscopy, enabling the localization of individual molecules even in densely labeled samples and in cellular and tissue environments. These microscopy techniques have been used to examine the organization and dynamics of plasma membrane components, providing insight into the fundamental interactions that determine membrane functions. Here, we broadly introduce the structure and organization of the mammalian plasma membrane and review recent applications of super-resolution microscopy to the study of membranes. We then highlight some inherent challenges faced when using super-resolution microscopy to study membranes, and we discuss recent technical advancements that promise further improvements to super-resolution microscopy and its application to the plasma membrane.
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http://dx.doi.org/10.1021/acs.chemrev.6b00716 | DOI Listing |
J Biomed Opt
December 2024
University of Strathclyde, Strathclyde Institute of Pharmacy and Biomedical Sciences, Glasgow, United Kingdom.
Significance: Current super-resolution imaging techniques allow for a greater understanding of cellular structures; however, they are often complex or only have the ability to image a few cells at once. This small field of view (FOV) may not represent the behavior across the entire sample, and manual selection of regions of interest (ROIs) may introduce bias. It is possible to stitch and tile many small ROIs; however, this can result in artifacts across an image.
View Article and Find Full Text PDFBrain
December 2024
Neuroimmunology Program, Fundació Clínic per la Recerca Biomèdica - Institut d'Investigacions Biomèdiques August Pi i Sunyer (FCRB-IDIBAPS), Barcelona 08036, Spain.
Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a disorder mediated by autoantibodies against the GluN1 subunit of NMDAR. It occurs with severe neuropsychiatric symptoms that often improve with immunotherapy. Clinical studies and animal models based on patients' antibody transfer or NMDAR immunization suggest that the autoantibodies play a major pathogenic role.
View Article and Find Full Text PDFLocalization microscopy enables imaging with resolutions that surpass the conventional optical diffraction limit. Notably, the Maximally INFormative LUminescence eXcitation (MINFLUX) method achieves super-resolution by shaping the excitation point spread function (PSF) to minimize the required photon flux for a given precision. Various beam shapes have recently been proposed to improve localization efficiency, yet their optimality remains an open question.
View Article and Find Full Text PDFPhilos Trans A Math Phys Eng Sci
December 2024
Institute of Optics, University of Rochester, Rochester, NY 14627, USA.
Can a quantum advantage for imaging resolution be realized with the help of quantum estimation theory? We expect so, but we show that, presently, theoretical tools are insufficiently developed to answer this question for extended objects. Still, there is much to be learned from the current state of the art. In this review, we re-examine prominent results in the literature and probe the limits of quantum metrology in addressing imaging resolution.
View Article and Find Full Text PDFAdv Sci (Weinh)
December 2024
Department of Bioengineering, Department of Electrical and Computer Engineering, Beckman Institute for Advanced Science and Technology, Cancer Center at Illinois, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA.
High-resolution optical microscopy, particularly super-resolution localization microscopy, requires precise real-time drift correction to maintain constant focus at nanoscale precision during the prolonged data acquisition. Existing methods, such as fiducial marker tracking, reflection monitoring, and bright-field image correlation, each provide certain advantages but are limited in their broad applicability. In this work, a versatile and robust drift correction technique is presented for single-molecule localization-based super-resolution microscopy.
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