Title:
光変換可能な光学標識を用いる光学顕微鏡法
Document Type and Number:
Japanese Patent JP4709278
Kind Code:
B2
Abstract:
A method for providing an image of a sample, the method comprising:
(a) providing activation radiation to a sample that includes phototransformable optical labels ("PTOLs"), the PTOLs being distributed in at least a portion of the sample with a density greater than an inverse of the diffraction-limited resolution volume of imaging optics, and the PTOLs being transformable from an inactivated state wherein the labels radiate little, if any, radiation at a wavelength characteristic of an activated and excited PTOL when exposed to excitation radiation, to an activated state wherein illumination by excitation radiation results in detectable fluorescence emission of radiation, to activate a first subset of the PTOLs in the sample;
(b) providing deactivation radiation to the sample to transform activated PTOLs to an unactivated state, wherein the deactivation radiation has a spatially-structured radiation field including intensity minima, such that a second subset of PTOLs located substantially at the minima of the deactivating radiation remain activated, while activated PTOLs exposed to the deactivating radiation outside the minima are substantially transformed into an unactivated form;
(c) providing excitation radiation to the sample to excite at least a portion of the activated PTOLs in the sample;
(d) controlling the intensity of the first activation radiation and controlling at least one of the intensity and the spatial structure of the deactivation radiation such that radiation emitted from activated and excited PTOLs located at the minima of the deactivation radiation in the sample is emitted substantially from at least one volume that is comparable to or less than a diffraction-limited resolution volume ("DLRV") of the imaging optics;
(e) detecting fluorescence radiation emitted from the activated and excited PTOLs with the imaging optics and determining the locations of the activated and excited PTOLs with superresolution accuracy;
(f) repeating (a) - (e), wherein the minima of the of the deactivating radiation are at different locations in the sample during different repetitions of (b); and
(g) combining the determined locations of the PTOLs from the different images to build up a superresoluiton image of the sample.
Inventors:
Harald F. Jesus
Robert E. Betjig
Robert E. Betjig
Application Number:
JP2008513618A
Publication Date:
June 22, 2011
Filing Date:
May 23, 2006
Export Citation:
Assignee:
Harald F. Jesus
Robert E. Betjig
Robert E. Betjig
International Classes:
G02B21/00; G01N21/64; G02B21/06; G02B21/36
Domestic Patent References:
| JP2004521323A | ||||
| JP2003015048A | ||||
| JP2001272343A |
Other References:
HELL S W, JAKOBS S, KASTRUP L,Imaging and writing at the nanoscale with focused visible light through saturable optical transitions,APPLIED PHYSICS A,2003年 8月 1日,vol. 77,pages 859-860
HELL S W ET AL,Concepts for nanoscale resolution in fluorescence microscopy,CURRENT OPINION IN NEUROBIOLOGY,2004年10月,vol. 14, no. 5,pages 599-609
ANDO R ET AL,Regulated Fast Nucleocytoplasmic Shuttling Observed by Reversible Protein Highlighting,SCIENCE,2004年11月19日,vol. 306, no. 5700,pages 1370-1373
深野天、宮脇敦史,理化学研究所 脳科学総合研究センター 細胞機能探索技術開発チームの研究開発,光技術コンタクト,日本,2003年11月20日,第41巻、第11号,第684頁-第688頁
HELL S W ET AL,Concepts for nanoscale resolution in fluorescence microscopy,CURRENT OPINION IN NEUROBIOLOGY,2004年10月,vol. 14, no. 5,pages 599-609
ANDO R ET AL,Regulated Fast Nucleocytoplasmic Shuttling Observed by Reversible Protein Highlighting,SCIENCE,2004年11月19日,vol. 306, no. 5700,pages 1370-1373
深野天、宮脇敦史,理化学研究所 脳科学総合研究センター 細胞機能探索技術開発チームの研究開発,光技術コンタクト,日本,2003年11月20日,第41巻、第11号,第684頁-第688頁
Attorney, Agent or Firm:
Toru Ishikawa
Hisako Ishido
Hisako Ishido