169 / 2024-06-14 11:09:05

Multifocal Structured illumination Microscopy based on Autocorrelation with Two-step Deconvolution

Super-resolution,MSIM,SOFI,SACD

Multifocal Structured Illumination Microscopy (MSIM) significantly enhances imaging capabilities, achieving a twofold increase in resolution compared to traditional widefield imaging. This technique is particularly advantageous for three-dimensional imaging of thick samples. However, the resolution improvement of MSIM is inherently limited by its imaging principle. To address this limitation, Super-resolution Optical Fluctuation Imaging (SOFI) can be combined with MSIM, offering a fourfold increase in resolution over widefield imaging. Yet, the SOFI method requires capturing hundreds of frames to produce a single image, necessitating high anti-bleaching properties of samples and resulting in longer acquisition times. We developed a method that integrates super-resolution imaging based on autocorrelation with two-step deconvolution (SACD) with MSIM, termed SACD-MSIM. By imaging samples labeled with photoblinking InP/ZnSe/ZnS core-shell quantum dot fluorescent probes, SACD-MSIM not only achieves a fourfold resolution improvement compared to widefield imaging but also dramatically reduces the time required for image acquisition. The introduction of SACD-MSIM enables more efficient and accurate three-dimensional super-resolution imaging of thick samples while preserving the integrity of the samples. This advancement is significant for the progression of three-dimensional MSIM imaging, providing a powerful tool for detailed structural analysis in complex biological specimens. The ability to perform high-resolution imaging swiftly and reliably makes SACD-MSIM a development in the field of super-resolution microscopy.