Abstract
The localization of point sources in optical microscopy enables nm-precision imaging of single-molecules and biological dynamics. We report a new method of localization microscopy using twin Airy beams that yields precise 3D localization with the key advantages of extended depth range, higher optical throughput, and potential for imaging higher emitter densities than are possible using other techniques. A precision of better than 30nm was achieved over a depth range in excess of 7 μm using a 60×, 1.4 NA objective. An illustrative application to extended-depth-range blood-flow imaging in a live zebrafish is also demonstrated.
Highlights
The localization of point sources in optical microscopy enables nm-precision imaging of singlemolecules and biological dynamics
We report a new method of localization microscopy using twin Airy beams that yields precise 3D localization with the key advantages of extended depth range, higher optical throughput, and potential for imaging higher emitter densities than are possible using other techniques
The precise localization of point emitters is of fundamental interest in biomedical research including superresolution (SR) microscopy [1,2], single-molecule tracking (SMT) in living cells [3,4], traction-force microscopy (TFM) [5,6], and in vivo flow measurement [7,8,9]
Summary
Yongzhuang Zhou , Paul Zammit, Vytautas Zickus, Jonathan M. We report a new method of localization microscopy using twin Airy beams that yields precise 3D localization with the key advantages of extended depth range, higher optical throughput, and potential for imaging higher emitter densities than are possible using other techniques. Airy beams yield accelerating lateral translation [14,15,16], which has potential for encoding 3D particle locations [9,12,17,18] Their “diffraction-free” propagation gives rise to an extended depth range and this property has been utilized for light sheet microscopy to increase the field of view [19]. Jia et al reported a modified version of an Airy beam for 3D particle localization microscopy, namely the self-bending PSF (SB-PSF) [12] This approach suffers from a reduced optical throughput, due to the truncation of the pupil. The TA-PSF exhibits a quasiuniform sensitivity to defocus over the whole depth range: it does not exhibit the reduced sensitivity of the Airy-beam PSF near the focus, nor the sign ambiguity
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