Journal ArticleDOI
Superresolution live imaging of plant cells using structured illumination microscopy
TLDR
A validated method for time-lapse SIM that focuses on cortical microtubules of different plant cell types by using one of the existing commercially available SIM platforms is described, which may be widely applied to the imaging of plant cells.Abstract:
This protocol describes a detailed method for superresolution imaging of plant tissues by structured illumination microscopy (SIM). Details include microscope calibration, tissue preparation, image acquisition and evaluation of SIM images. Although superresolution (SR) approaches have been routinely used for fixed or living material from other organisms, the use of time-lapse structured illumination microscopy (SIM) imaging in plant cells still remains under-developed. Here we describe a validated method for time-lapse SIM that focuses on cortical microtubules of different plant cell types. By using one of the existing commercially available SIM platforms, we provide a user-friendly and easy-to-follow protocol that may be widely applied to the imaging of plant cells. This protocol includes steps describing calibration of the microscope and channel alignment, generation of an experimental point spread function (PSF), preparation of appropriate observation chambers with available plant material, image acquisition, reconstruction and validation. This protocol can be carried out within two to three working days.read more
Citations
More filters
Methods and Applications
Ajit Varki,Richard D Cummings,Jeffrey D. Esko,Hudson Freeze,Pamela Stanley,Carolyn R. Bertozzi,Gerald W. Hart,Marilynn E. Etzler +7 more
TL;DR: The aim of the research presented in this thesis is to create new methods for design for manufacturing, by using several approaches of KE, and find the beneficial and less beneficial aspects of these methods in comparison to each other and earlier research.
Journal ArticleDOI
Super-resolution microscopy demystified
Lothar Schermelleh,Alexia Ferrand,Thomas R Huser,Christian Eggeling,Markus Sauer,Oliver Biehlmaier,Drummen Gpc. +6 more
TL;DR: An overview of current super-resolution microscopy techniques is given and guidance on how best to use them to foster biological discovery is provided.
Journal ArticleDOI
Strategic and practical guidelines for successful structured illumination microscopy
Justin Demmerle,Cassandravictoria Innocent,Alison J. North,Graeme Ball,Marcel Müller,Ezequiel Miron,Atsushi Matsuda,Atsushi Matsuda,Ian M. Dobbie,Yolanda Markaki,Lothar Schermelleh +10 more
TL;DR: This protocol allows users to generate high-quality SIM data while accounting and correcting for common artifacts, and allows researchers from students to imaging professionals to create an optimal SIM imaging environment regardless of specimen type or structure of interest.
Journal ArticleDOI
Imaging cellular structures in super-resolution with SIM, STED and Localisation Microscopy: A practical comparison.
Eva Wegel,Antonia Gohler,B C Lagerholm,Alan Wainman,Stephan Uphoff,Rainer Kaufmann,Rainer Kaufmann,Ian M. Dobbie +7 more
TL;DR: In this article, the authors compare the individual strengths and weaknesses of SIM, STED, and SMLM by imaging a variety of different subcellular structures in fixed cells.
Journal ArticleDOI
Unconventional methods of imaging: computational microscopy and compact implementations.
Euan McLeod,Aydogan Ozcan +1 more
TL;DR: A review of lens-based computational imaging, in particular, light-field microscopy, structured illumination, synthetic aperture, Fourier ptychography, and compressive imaging, and how these and other microscopy modalities have been implemented in compact and field-portable devices, often based around smartphones.
References
More filters
Journal ArticleDOI
Fiji: an open-source platform for biological-image analysis
Johannes Schindelin,Ignacio Arganda-Carreras,Erwin Frise,Verena Kaynig,Mark Longair,Tobias Pietzsch,Stephan Preibisch,Curtis Rueden,Stephan Saalfeld,Benjamin Schmid,Jean-Yves Tinevez,Daniel J. White,Volker Hartenstein,Kevin W. Eliceiri,Pavel Tomancak,Albert Cardona +15 more
TL;DR: Fiji is a distribution of the popular open-source software ImageJ focused on biological-image analysis that facilitates the transformation of new algorithms into ImageJ plugins that can be shared with end users through an integrated update system.
Journal ArticleDOI
Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy.
TL;DR: Lateral resolution that exceeds the classical diffraction limit by a factor of two is achieved by using spatially structured illumination in a wide‐field fluorescence microscope with strikingly increased clarity compared to both conventional and confocal microscopes.
Journal ArticleDOI
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
TL;DR: Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples, and a recently proposed method in which the nonlinearity arises from saturation of the excited state is experimentally demonstrated.
Book
Methods and applications
TL;DR: A survey of sampling principles can be found in this paper, where the authors present sample designs for some common sampling problems such as bias and non-sampling errors in survey results.
Journal ArticleDOI
A guide to super-resolution fluorescence microscopy
TL;DR: These new super-resolution technologies are either based on tailored illumination, nonlinear fluorophore responses, or the precise localization of single molecules and have created unprecedented new possibilities to investigate the structure and function of cells.