Live cell single molecule-guided Bayesian localization super resolution microscopy
Dear Editor,Many current super resolution (SR) microscopic techniques ”1-6” have been successfully applied to image cellular dynamics in living cells,but their applications have remained technically challenging.Live cell stimulated emission depletion (STED)/reversible saturable optical linear fluorescence transitions (RESOLFT) microscopy and structured illumination microscopy (SIM)/nonlinear SIM require sophisticated expensive optical setups and specialized expertise for accurate optical alignment.Live cell photo-activated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM)use less complicated setup;however,a scientific complementary metal-oxide-semiconductor (sCMOS) camera,whose pixel-dependent noise must be characterized and calibrated before use ”7”,is required for extremely high acquisition speed over tens of thousands of frames.Recently,wide field-based SR microscopies have been developed to improve temporal resolution using much fewer time-lapse images (hundreds to thousands) than PALM/STORM ”8,9”.One of them,Bayesian analysis of the blinking and bleaching (3B) ”8”,offers enormous potential to resolve ultrastructure and fast cellular dynamics in living cells beyond the diffraction limit.Despite its potential,3B analysis is impractical when imaging the nanoscale dynamics in large fields of view over long time periods,as the calculation is extremely time-consuming ”8” and/or the analysis consumes large amounts of web resources ”10”.Another major problem of 3B imaging is the artificial thinning and thickening of structures both in simulated image data and in experimental data ”8,10”.
stimulated emission、experimental data、diffraction limit、super resolution、web resources、major problem
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O4 ;TP3
the National Basic Research Program2013 CB910103;the National Key Research and Development Projects2016YFA0501500;the National Natural Science Foundation of China31421002,31370851,31170818,31300612,61232001,61472397,61672493 and 61502455;Project of the Chinese Academy of SciencesXDB08030202;Beijing Natural Science Foundation7131011;Youth Innovation Promotion Association CAS2012080 and 2013067;Young Elite Scientist Sponsorship Program by CAST
2017-06-08（万方平台首次上网日期，不代表论文的发表时间）
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