Format

Send to

Choose Destination
Opt Express. 2012 Feb 27;20(5):4957-67. doi: 10.1364/OE.20.004957.

PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking.

Author information

1
Laboratoire Kastler Brossel, CNRS UMR 8552, Département de Physique et Institut de Biologie de l’Ecole Normale Supérieure, Université Pierre et Marie Curie-Paris6, 46 rue d’Ulm 75230 Paris cedex 05, France.

Abstract

We present a novel approach for three-dimensional localization of single molecules using adaptive optics. A 52-actuator deformable mirror is used to both correct aberrations and induce two-dimensional astigmatism in the point-spread-function. The dependence of the z-localization precision on the degree of astigmatism is discussed. We achieve a z-localization precision of 40 nm for fluorescent proteins and 20 nm for fluorescent dyes, over an axial depth of ~800 nm. We illustrate the capabilities of our approach for three-dimensional high-resolution microscopy with super-resolution images of actin filaments in fixed cells and single-molecule tracking of quantum-dot labeled transmembrane proteins in live HeLa cells.

PMID:
22418300
[Indexed for MEDLINE]

Supplemental Content

Loading ...
Support Center