Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function Export

@article{citeulike:4036416, abstract = {10.1073/pnas.0900245106 We demonstrate single-molecule fluorescence imaging beyond the optical diffraction limit in 3 dimensions with a wide-field microscope that exhibits a double-helix point spread function (DH-PSF). The DH-PSF design features high and uniform Fisher information and has 2 dominant lobes in the image plane whose angular orientation rotates with the axial () position of the emitter. Single fluorescent molecules in a thick polymer sample are localized in single 500-ms acquisitions with 10- to 20-nm precision over a large depth of field (2 μm) by finding the center of the 2 DH-PSF lobes. By using a photoactivatable fluorophore, repeated imaging of sparse subsets with a DH-PSF microscope provides superresolution imaging of high concentrations of molecules in all 3 dimensions. The combination of optical PSF design and digital postprocessing with photoactivatable fluorophores opens up avenues for improving 3D imaging resolution beyond the Rayleigh diffraction limit.}, author = {Pavani, Sri R. and Thompson, Michael A. and Biteen, Julie S. and Lord, Samuel J. and Liu, Na and Twieg, Robert J. and Piestun, Rafael and Moerner, W. E.}, citeulike-article-id = {4036416}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0900245106}, citeulike-linkout-1 = {http://www.pnas.org/content/0900245106v1//.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/0900245106v1//.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19211795}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19211795}, day = {11}, doi = {10.1073/pnas.0900245106}, journal = {Proceedings of the National Academy of Sciences}, keywords = {3d, dcdhf, microscopy, optics, super-resolution}, month = {February}, posted-at = {2009-02-11 19:09:57}, priority = {2}, title = {Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function}, url = {http://dx.doi.org/10.1073/pnas.0900245106}, year = {2009} }

TY - JOUR ID - citeulike:4036416 L3 - citeulike-article-id:4036416 N2 - 10.1073/pnas.0900245106 We demonstrate single-molecule fluorescence imaging beyond the optical diffraction limit in 3 dimensions with a wide-field microscope that exhibits a double-helix point spread function (DH-PSF). The DH-PSF design features high and uniform Fisher information and has 2 dominant lobes in the image plane whose angular orientation rotates with the axial () position of the emitter. Single fluorescent molecules in a thick polymer sample are localized in single 500-ms acquisitions with 10- to 20-nm precision over a large depth of field (2 μm) by finding the center of the 2 DH-PSF lobes. By using a photoactivatable fluorophore, repeated imaging of sparse subsets with a DH-PSF microscope provides superresolution imaging of high concentrations of molecules in all 3 dimensions. The combination of optical PSF design and digital postprocessing with photoactivatable fluorophores opens up avenues for improving 3D imaging resolution beyond the Rayleigh diffraction limit. JF - Proceedings of the National Academy of Sciences TI - Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function KW - 3d KW - dcdhf KW - microscopy KW - optics KW - super-resolution AU - Pavani, Sri AU - Thompson, Michael AU - Biteen, Julie AU - Lord, Samuel AU - Liu, Na AU - Twieg, Robert AU - Piestun, Rafael AU - Moerner, WE PY - 2009/02/11/ UR - http://dx.doi.org/10.1073/pnas.0900245106 ER -