Super Resolution Microscopy
Super Resolution Microscopy - Related Videos
16:52
Test Samples for Optimizing STORM Super-Resolution Microscopy
0 Views •
Cited by 48 •
2013
We describe the preparation of three test samples and how they can be used to optimize and assess the performance of STORM microscopes. Using these examples we show how to acquire raw data and then process it to acquire super-resolution images in cells of approximately 30-50 nm resolution.
01:37
Super-resolution Fluorescence Microscopy
0 Views •
2023
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
12:28
Correlative Super-resolution and Electron Microscopy to Resolve Protein Localization in Zebrafish Retina
0 Views •
Cited by 11 •
2017
This protocol describes the necessary steps to obtain subcellular protein localization results on zebrafish retina by correlating super-resolution light microscopy and scanning electron microscopy images.
07:38
Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
0 Views •
Cited by 9 •
2017
This manuscript uses the Fiji-based open-source software package VirusMapper to apply single-particle analysis to super-resolution microscopy images in order to generate precise models of nanoscale structure.
11:57
Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)
0 Views •
Cited by 10 •
2016
We present a protocol for the application of interferometric PhotoActivated Localization Microscopy (iPALM), a 3-dimensional single-molecule localization super resolution microscopy method, to the imaging of the actin cytoskeleton in adherent mammalian cells. This approach allows light-based visualization of nanoscale structural features that would otherwise remain unresolved by conventional diffraction-limited optical microscopy.
FAQs
Single-molecule localization methods control fluorescence in time by switching individual fluorophores on and off. This prevents nearby emitters from appearing simultaneously as one unresolved signal, allowing their positions to be examined separately. The resulting localization information reveals molecular organization that conventional light microscopy cannot distinguish, making it useful for studying protein distributions and cellular architecture.
Structured illumination microscopy controls fluorescence spatially rather than relying on sequentially switching individual fluorophores. Patterned illumination interacts with labeled structures to provide additional spatial information, which improves the visibility of cellular features beyond conventional optical resolution. This approach is especially relevant when researchers want to examine organized membranes, cytoskeletal networks, or organelles within biological samples.
Fluorescence control separates information that would otherwise overlap in an optical image. By regulating when or where fluorescent signals appear, these methods expose molecular arrangements hidden within conventional images. That added detail helps connect the distribution of proteins and other labeled components with the architecture of membranes, organelles, and cytoskeletal networks.
Applications include membranes, cytoskeletal networks, organelles, and protein distributions. Examining these features at nanoscale detail can show how cellular components are arranged rather than only indicating their presence in a broader region. Researchers can therefore investigate cell organization and relate molecular architecture to the functions carried out by specific cellular structures.
The methods can be applied to either fixed or living cells, so the choice depends on the biological question and the sample state being studied. Fixed-cell imaging supports examination of preserved cellular organization, whereas live-cell imaging allows structures to be investigated in living specimens. This distinction determines whether the experiment emphasizes structural detail or observations within living cells.
It is useful when disease-related changes involve molecular organization that remains unresolved with conventional light microscopy. By revealing protein distributions and cellular architecture at nanoscale detail, the methods can provide structural context for altered cell organization. This information supports research connecting molecular arrangements with disease mechanisms rather than treating cellular components as isolated signals.
Nanoscale images provide information about where proteins and other cellular components are distributed within membranes, organelles, or cytoskeletal networks. Researchers can use those spatial patterns to examine how molecular architecture relates to signaling and broader cellular function. The approach therefore links visual evidence of organization with biological questions about how cells are arranged and operate.