PALM/STORM and SPT Improve resolution and go 3D
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The quality of the Point Spread Function (PSF) is particularly important in single molecule localization microscopy (SMLM) methods such as PALM/STORM and SPT. By correcting both optical setup and sample-induced aberrations, adaptive optics optimizes the shape of the PSF, increasing the number of detected photons. This directly enhances the localization precision (resolution) for these methods. In addition to that, by applying a controllable amount of perfect astigmatism, adaptive optics enables the ultimate 3D imaging capability with excellent temporal stability. This pure astigmatism superimposed onto the corrected PSF delivers unprecedented accuracy for single molecule localization in all three dimensions. Control over aberrations allows the user to obtain the same PSF over the whole biological sample and enables unparalleled single particle tracking precision. For an easy implementation of adaptive optics in the microscope setup, Imagine Optic developed a plug and play solution: MicAO 3DSR.
Selected publications: O. Glushonkov et al. “Optimized protocol for combined PALM-dSTORM imaging.” Sci. Rep. 2018, 8, 8749. G. Clouvel et al. “Dual-color 3D PALM/dSTORM imaging of centrosomal proteins using MicAO 3DSR.” Proc. SPIE 2013, 8590, 85 900Z+. I. Izeddin et al. “PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking.” Opt. Express 2012, 20, 4957–4967.