Structured Illumination Microscopy
A simple overview

01

What is Structured Illumination Microscopy (SIM)?

Structured Illumination Microscopy is a technique that can double the resolution of images when compared with standard confocal microscopy. It is liked because it is easier to use than some alternative super-resolution techniques such as dSTORM.

The enhanced resolution and the high speed nature of SIM, makes it suitable for studies of subcellular structures in depth as well as time-series imaging in living cells. The detection configuration of the Nanoimager allows this powerful technique to be applied for dual-color simultaneous super-resolution imaging.

Structured Illumination Microscopy

Image credit - Surgical Photonics & Engineering Laboratory, Harvard Medical School, Boston (scale bar: 10µm)

02

What are the advantages of Structured Illumination Microscopy?

SIM microscopy works well with conventional fluorophores so imaging existing samples doesn't require a complete reinvention of fluorophore protocols. Therefore, getting a higher resolution than widefield microscopy only means using a more advanced microscope not reinventing sample preparation.

SIM also has greater imaging speeds than localization microscopy. It is thus more compatible with live-cell super-resolution applications. It is worth checking out the exact microscope methodology as some will bleach samples faster than others depending on the how frequently each part of the sample is illuminated by the lasers.

Optical sectioning (equivalent to confocal imaging) and super-resolution structured illumination microscopy are easily performed on thicker samples such as tissues.

SIM microscopy

Image credit - Prof. D. Jakimowicz lab, the University of Wroclaw, Poland (scale bar: 10µm)

03

What are the limitations of Structured Illumination Microscopy?

If you are after the ultimate sample resolutions (20nm to 50nm), then structured illumination microscopy is beaten by other techniques including dSTORM and PALM microscopy. In addition it is easy for artefacts to be generated from poor sample preparation so it is important to seek the best advice before starting.

Structured Illumination Microscope

Image credit - Dr. Cristiane Beninca, Light Microscopy Facility, MRC-MBU University of Cambridge (scale bar: 10µm)

04

Why should the Nanoimager be considered for SIM Microscopy?

The Nanoimager is capable of 4 color imaging, 2 colors simultaneously, and has an advanced z-stack set up to allow samples to be imaged at different depths. Live cell imaging is supported by time lapse capability and a design that supports microfluidics setups. With no post processing, images are ready once the acquisition is complete.

The Nanoimager is a very versatile microscope, capable of SIM microscopy, dSTORM, PALM, single-particle tracking and smFRET among other techniques. Its unique design means it has unrivalled stability and never needs aligning. Its robustness, coupled with it small size means that it can be employed in any lab location. There's no requirement for a dark room or an optical table.

As a SIM microscope, the Nanoimager capabilities have been employed in applications as diverse as mitochondria, tissue imaging, following cell division and cytoskeleton research.



Read more about the Nanoimager's super-resolution microscopy techniques and illumination modes.

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