scholarly journals Adaptive optics improves multiphoton super-resolution imaging

2017 ◽  
Vol 14 (9) ◽  
pp. 869-872 ◽  
Author(s):  
Wei Zheng ◽  
Yicong Wu ◽  
Peter Winter ◽  
Robert Fischer ◽  
Damian Dalle Nogare ◽  
...  
2012 ◽  
Vol 20 (5) ◽  
pp. 4957 ◽  
Author(s):  
Ignacio Izeddin ◽  
Mohamed El Beheiry ◽  
Jordi Andilla ◽  
Daniel Ciepielewski ◽  
Xavier Darzacq ◽  
...  

2021 ◽  
Author(s):  
Sanghyeon Park ◽  
Yonghyeon Jo ◽  
Minsu Kang ◽  
Jin Hee Hong ◽  
Sangyoon Ko ◽  
...  

Specimen-induced aberration has been one of the major factors limiting the imaging depth in single-molecule localization microscopy (SMLM). In this study, we measured the wavefront of intrinsic reflectance signal at the fluorescence emission wavelength to construct a time-gated reflection matrix and find complex tissue aberration without resorting to fluorescence detection. Physically correcting the identified aberration via wavefront shaping with a liquid-crystal spatial light modulator (SLM) enables super-resolution imaging even when the aberration is too severe for initiating localization processes. We demonstrate the correction of complex tissue aberration, the root-mean-square (RMS) wavefront distortion of which is more than twice the 1 rad limit presented in previous studies; this leads to the recovery of single molecules by 77 times increased localization number. We visualised dendritic spines in mouse brain tissues and early myelination processes in a whole zebrafish at up to 102 μm depth with 28-39 nm localization precision. The proposed approach can expand the application range of SMLM to thick samples that cause the loss of localization points owing to severe aberration.


2019 ◽  
Author(s):  
Piotr Zdankowski ◽  
Maciej Trusiak ◽  
David McGloin ◽  
Jason R. Swedlow

AbstractIn stimulated emission depletion (STED) nanoscopy, the major origin of decreased signal-to-noise ratio within images can be attributed to sample photobleaching and strong optical aberrations. This is due to STED utilising both a high power depletion laser (increasing risk of photodamage), while the depletion beam is very sensitive to sample-induced aberrations. Here we demonstrate a custom-built 3D STED microscope with automated aberration correction that is capable of 3D super-resolution imaging through thick, highly aberrating, tissue. We introduce and investigate image denoising by block-matching and collaborative filtering (BM3D) to numerically enhance fine object details otherwise mixed with noise. Numerical denoising provides an increase in the final effective resolution of the STED imaging of 31% using the well-established Fourier ring correlation metric. Experimental validation of the proposed method is achieved through super-resolved 3D imaging of axons in differentiated induced pluripotent stem cells growing under a 80µm thick layer of tissue with lateral and axial resolution of 256nm and 300nm, respectively.


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