Enhancing multi-spot structured illumination microscopy with fluorescence difference

• Main Authors: Edward N. Ward, Frida H. Torkelsen, Robert Pal
• Format: Article
• Language: English
• Published: The Royal Society 2018-01-01
• Series: Royal Society Open Science
• Subjects: super-resolution microscopy; structured illumination; msim; difference microscopy
• Online Access: https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.171336

Structured illumination microscopy is a super-resolution technique used extensively in biological research. However, this technique is limited in the maximum possible resolution increase. Here we report the results of simulations of a novel enhanced multi-spot structured illumination technique. This method combines the super-resolution technique of difference microscopy with structured illumination deconvolution. Initial results give at minimum a 1.4-fold increase in resolution over conventional structured illumination in a low-noise environment. This new technique also has the potential to be expanded to further enhance axial resolution with three-dimensional difference microscopy. The requirement for precise pattern determination in this technique also led to the development of a new pattern estimation algorithm which proved more efficient and reliable than other methods tested.