Multifocus structured illumination microscopy for fast volumetric super-resolution imaging

Abstract

Abstract:We here report for the first time the synergistic implementation of structured illumination microscopy (SIM) and multifocus microscopy (MFM). This imaging modality is designed to alleviate the problem of insufficient volumetric acquisition speed in superresolution biological imaging. SIM is a wide-field super-resolution technique that allows imaging with visible light beyond the classical diffraction limit. Employing multifocus diffractive optics we obtain simultaneous wide-field 3D imaging capability in the SIM acquisition sequence, improving volumetric acquisition speed by an order of magnitude. Imaging performance is demonstrated on biological specimens.

In Situ Encapsulation of Nile Red or Doxorubicin during RAFT‐Mediated Emulsion Polymerization via Polymerization‐Induced Self‐Assembly for Biomedical Applications

“…The successful loading of the NR into the hydrophobic core of the PDMAPMA-b-PMMA NR NPs was further verified using structured illumination microscopy (SIM), Figure 2a. [28] Grey circles were observed in the corresponding micrograph, indicating that the hydrophobic interior of the NPs is loaded with NR. The sizes of the NPs were found to be ≈200 nm, Figure 2a.…”

Correlative cryo-structured illumination fluorescence microscopy and soft X-ray tomography elucidates reovirus intracellular release pathway

“…(b) it can image relatively thick samples, reducing the need for sample sectioning prior to imaging; (c) it uses relatively low light doses (10-100 W/cm 2 laser power) and short image acquisition times (20-100ms per single exposure), minimising sample heating and reducing the chance of ice crystal formation which would otherwise lead to sample damage and image quality degradation 35 and, (d) it has extremely good out of focus light suppression, leading to high contrast images even in samples with thicknesses of 10µm or more 36 .…”

“…Samples have to be kept below the glass transition temperature of circa 130°K (-140°C), whilst mounted close to an objective that is kept at room temperature and while at the same time being illuminated with high intensity light for excitation purposes. Given these challenges, 3D-structure illumination microscopy 34,35 is the natural choice for a super resolution method for cryogenically preserved biological samples as it can deliver fast 3D super-resolution imaging with relatively low exposures to laser light and wide fields of illumination.…”

High speed multi-plane super-resolution structured illumination microscopy of living cells using an image-splitting prism

“…Chromatic aberrations can be corrected using additional elements [12]. Recently, this approach was combined with a commercial SR-SIM microscope [13] and provided the first multi-plane SR-SIM images in a prototype system. Although a great demonstration of the future of SR-SIM imaging, these tests ended up being limited by the slow speed of the commercial SR-SIM platform, which was not originally conceived for video-rate super-resolution imaging.…”