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Related Experiment Video

Updated: Jun 16, 2026

Single Plane Illumination Module and Micro-capillary Approach for a Wide-field Microscope

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Published on: August 15, 2014

Strategic and practical guidelines for successful structured illumination microscopy.

Justin Demmerle¹, Cassandravictoria Innocent¹, Alison J North²

¹Micron Advanced Bioimaging Unit, Department of Biochemistry, University of Oxford, Oxford, UK.

Nature Protocols

|April 14, 2017

Summary

This summary is machine-generated.

Structured Illumination Microscopy (SIM) provides high-resolution 3D imaging but can suffer from artifacts. This protocol details artifact correction and calibration for high-quality SIM data, saving time and resources.

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Last Updated: Jun 16, 2026

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Area of Science:

Microscopy
Biotechnology
Optical Imaging

Background:

Structured Illumination Microscopy (SIM) offers subdiffraction resolution for multicolor 3D imaging of biological specimens.
SIM's reliance on post-processing makes it susceptible to artifacts that can degrade image quality and interpretation.
Artifacts in SIM can lead to reduced resolution, compromised data, and increased experimental costs.

Purpose of the Study:

To present a protocol for generating high-quality SIM data by addressing common artifacts.
To provide researchers with methods for artifact correction and system calibration in SIM experiments.
To enable optimal SIM imaging environments for diverse specimens and research goals.

Main Methods:

Development of a protocol for preparing calibration bead slides specifically for SIM.
Detailed procedures for acquiring calibration data and documenting common SIM artifacts.
Inclusion of corrective measures for identified artifacts and a checklist for experimental design.

Main Results:

The protocol facilitates the generation of high-quality SIM data by accounting for and correcting artifacts.
Calibration sample preparation and system calibration can be completed within 1-2 days.
The protocol is applicable to various commercially available or custom SIM platforms.

Conclusions:

This protocol empowers researchers, from students to professionals, to optimize their SIM imaging.
It ensures reliable and interpretable SIM data by mitigating common artifacts.
The standardized approach enhances the efficiency and accuracy of super-resolution microscopy.