Structured Illumination Microscopy
Structured Illumination Microscopy - Related Videos
11:15
A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
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Cited by 114 •
2016
This article provides an in depth guide for the assembly and operation of a structured illumination microscope operating with total internal reflection fluorescence illumination (TIRF-SIM) to image dynamic biological processes with optical super-resolution in multiple colors.
14:11
Imaging Dendritic Spines of Rat Primary Hippocampal Neurons using Structured Illumination Microscopy
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Cited by 10 •
2014
This article describes a working protocol to image dendritic spines from hippocampal neurons in vitro using Structured Illumination Microscopy (SIM). Super-resolution microscopy using SIM provides image resolution significantly beyond the light diffraction limit in all three spatial dimensions, allowing the imaging of individual dendritic spines with improved detail.
10:07
Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers
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2014
High-resolution intravital imaging with enhanced contrast up to 120 µm depth in lymph nodes of adult mice is achieved by spatially modulating the excitation pattern of a multi-focal two-photon microscope. In 100 µm depth we measured resolutions of 487 nm (lateral) and 551 nm (axial), thus circumventing scattering and diffraction limits.
12:44
Super-resolution Imaging of the Cytokinetic Z Ring in Live Bacteria Using Fast 3D-Structured Illumination Microscopy (f3D-SIM)
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Cited by 22 •
2014
Spatiotemporal information about dynamic proteins inside live cells is crucial for understanding biology. A type of super-resolution microscopy called fast 3D-structured illumination microscopy (f3D-SIM) reveals unique information about the cytokinetic Z ring in bacteria: both its bead-like appearance and the rapid dynamics of FtsZ within the ring.
09:09
Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data
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Cited by 17 •
2015
To investigate flow velocities and directionality of filamentous-actin at the T cell immunological synapse, live-cell super-resolution imaging is combined with total internal reflection fluorescence and quantified with spatio-temporal image correlation spectroscopy.
FAQs
The projected stripes create interference patterns that encode spatial information from the specimen. A computational reconstruction then combines this information to produce an image with improved resolution compared with a conventional light microscope. The key principle is that illumination and image processing work together, allowing cellular features beyond the usual diffraction limit to be visualized.
Conventional light microscopy is limited by diffraction, which restricts how closely spaced cellular features can appear as separate structures. SIM adds known patterned illumination to generate interference patterns containing additional spatial information. Combining those data computationally improves the reconstructed image, making organelles, cytoskeletal features, and molecular distributions easier to examine.
A relatively low illumination dose helps SIM examine living cells while limiting the amount of light delivered during observation. This feature is especially relevant when researchers need to preserve temporal information across rapid sequences. It supports studies of dynamic biology, including trafficking, cell division, and signaling, rather than restricting imaging to fixed specimens.
A typical workflow begins by illuminating the specimen with a known striped or otherwise patterned light field. The resulting interference information is recorded as image data, which are then computationally combined to reconstruct an improved-resolution image. This sequence links controlled illumination with image processing and can be applied to fixed or living cells.
The technique can visualize organelles, cytoskeletal features, and molecular distributions in biological specimens. Its use is not limited to fixed preparations; it can also support imaging of living cells. That flexibility allows researchers to study both cellular organization and changes in that organization over time, depending on the biological question.
Biologists may choose SIM when they need improved spatial resolution while also following events over time. Its relatively low illumination dose and ability to capture rapid sequences support investigations of trafficking, cell division, and signaling. The resulting images help connect the location of cellular structures with dynamic processes while retaining temporal information.