title: Live imaging using adaptive optics with fluorescent protein guide-stars.
abstract: Spatially and temporally dependent optical aberrations induced by the inhomogeneous refractive index of live samples limit the resolution of live dynamic imaging. We introduce an adaptive optical microscope with a direct wavefront sensing method using a Shack-Hartmann wavefront sensor and fluorescent protein guide-stars for live imaging. The results of imaging Drosophila embryos demonstrate its ability to correct aberrations and achieve near diffraction limited images of medial sections of large Drosophila embryos. GFP-polo labeled centrosomes can be observed clearly after correction but cannot be observed before correction. Four dimensional time lapse images are achieved with the correction of dynamic aberrations. These studies also demonstrate that the GFP-tagged centrosome proteins, Polo and Cnn, serve as excellent biological guide-stars for adaptive optics based microscopy.
authors: W.M. Keck Center for Adaptive Optical Microscopy, Jack Baskin School of Engineering, University of California, Santa Cruz, California 95064, USA. taoxd@soe.ucsc.edu
authors: Xiaodong
authors: X
authors: Tao
authors: Justin
authors: J
authors: Crest
authors: Shaila
authors: S
authors: Kotadia
authors: Oscar
authors: O
authors: Azucena
authors: Diana C
authors: DC
authors: Chen
authors: William
authors: W
authors: Sullivan
authors: Joel
authors: J
authors: Kubby
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000196:embryology; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence; D055095:Optics and Photonics / Q000379:methods*; D013997:Time Factors; D058067:Wavelet Analysis
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins