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The development of real-time single-molecule detection and tracking technology in time and space for multiple bio-molecules in intact cells has important significance for the study on molecular behaviors in life processes. However, limited depth of field is the main drawback of conventional microscopy that prevents observation and tracking of multiple molecules in intact cells. Based on the principle of wavefront coding, the design and fabrication of a novel diffractive optical element (DOE) which combines a distorted diffraction grating with a double-helix point spread function (PSF) phase plate so that it can simultaneously perform multi-plane imaging with double-helix PSF and realize an extended depth of field is presented. Experiments have been carried out on a self-built microscopic system based on the novel DOE, showing that a depth of field can be up to 12 μm. Experimental results are in good agreement with the theoretical predictions, thus proving the feasibility of this method.
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Keywords:
- multi-molecule tracking /
- diffractive optical element /
- distorted diffraction grating /
- double helix point spread function
[1] Betzig E, Patterson G H, Sougrat R, Lindwasser O W, Olenych S, Bonifacino J S, Davidson M W, Lippincott-Schwartz J, Hess H F 2006 Science 313 1642
[2] Rust M J, Bates M, Zhuang X 2006 Nat. Methods 3 793
[3] Hess S T, Girirajan T P K, Mason M D 2006 Bio. Phys. J 91 4258
[4] Xie X S, Yu J, Yang W Y 2006 Science 312 228
[5] Chen D N, Liu L, Yu B, Niu H B 2010 Acta Phys. Sin. 59 6948 (in Chinese) [陈丹妮, 刘磊, 于斌, 牛憨笨 2010 物理学报 59 6948]
[6] Herbert S, Soares H, Zimmer C, Henriques R 2012 Microsc. Microanal. 18 1419
[7] Jones S A, Shim S H, He J, Zhuang X 2011 Nat. Methods 8 499
[8] Ober R J, Ram S, Ward E S 2004 Bio. Phys. J 86 1185
[9] Dupont A, Lamb D C 2011 Nanoscale 3 4532
[10] Yildiz A, Tomishige M, Vale R D, Selvin P R 2004 Science 303 676
[11] Arhel N, Genovesio A, Kim K A, Miko S, Perret E, Olivo-Marin J C, Shorte S, Charneau P 2006 Nat. Methods 3 817
[12] Speidel M, Jonas A, Florin E L 2003 Opt. Lett. 28 69
[13] Levi V, Ruan Q Q, Gratton E 2005 Bio. Phys. J. 88 2919
[14] Park Y, Popescu G, Badizadegan K, Dasari R R, Feld M S 2007 Opt. Lett. 32 811
[15] Kao H P, Verkman A S 1994 Bio. Phys. J. 67 1291
[16] Pavani S R, Piestun R 2008 Opt. Express 16 22048
[17] Ram S, Prabhat P, Chao J, Ward E S, Ober R J 2008 Bio. Phys. J. 95 6025
[18] Piestun R, Schechner Y Y, Shamir J 2000 J. Opt. Soc. Am. A 17 294
[19] Liu M, Chen X Y, Li H X, Song H S, Teng S Y, Cheng C F 2010 Acta Phys. Sin. 59 8490 (in Chinese) [刘曼, 陈小艺, 李海霞, 宋洪胜, 滕树云, 程传福 2010 物理学报 59 8490]
[20] Pavani S R P, Greengard A, Piestun R 2009 Appl. Phys. Lett. 95 021103
[21] Blanchard M P, H Greenaway A 1999 Appl. Opt. 38 6692
[22] Li H, Yu B, Chen D N, Niu H B 2013 Acta Phys. Sin. 62 124201 (in Chinese) [李恒, 于斌, 陈丹妮, 牛憨笨 2013 物理学报 62 124201]
[23] Wu C, Chiu D T 2013 Angew. Chem. Int. Ed. 11 3086
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[1] Betzig E, Patterson G H, Sougrat R, Lindwasser O W, Olenych S, Bonifacino J S, Davidson M W, Lippincott-Schwartz J, Hess H F 2006 Science 313 1642
[2] Rust M J, Bates M, Zhuang X 2006 Nat. Methods 3 793
[3] Hess S T, Girirajan T P K, Mason M D 2006 Bio. Phys. J 91 4258
[4] Xie X S, Yu J, Yang W Y 2006 Science 312 228
[5] Chen D N, Liu L, Yu B, Niu H B 2010 Acta Phys. Sin. 59 6948 (in Chinese) [陈丹妮, 刘磊, 于斌, 牛憨笨 2010 物理学报 59 6948]
[6] Herbert S, Soares H, Zimmer C, Henriques R 2012 Microsc. Microanal. 18 1419
[7] Jones S A, Shim S H, He J, Zhuang X 2011 Nat. Methods 8 499
[8] Ober R J, Ram S, Ward E S 2004 Bio. Phys. J 86 1185
[9] Dupont A, Lamb D C 2011 Nanoscale 3 4532
[10] Yildiz A, Tomishige M, Vale R D, Selvin P R 2004 Science 303 676
[11] Arhel N, Genovesio A, Kim K A, Miko S, Perret E, Olivo-Marin J C, Shorte S, Charneau P 2006 Nat. Methods 3 817
[12] Speidel M, Jonas A, Florin E L 2003 Opt. Lett. 28 69
[13] Levi V, Ruan Q Q, Gratton E 2005 Bio. Phys. J. 88 2919
[14] Park Y, Popescu G, Badizadegan K, Dasari R R, Feld M S 2007 Opt. Lett. 32 811
[15] Kao H P, Verkman A S 1994 Bio. Phys. J. 67 1291
[16] Pavani S R, Piestun R 2008 Opt. Express 16 22048
[17] Ram S, Prabhat P, Chao J, Ward E S, Ober R J 2008 Bio. Phys. J. 95 6025
[18] Piestun R, Schechner Y Y, Shamir J 2000 J. Opt. Soc. Am. A 17 294
[19] Liu M, Chen X Y, Li H X, Song H S, Teng S Y, Cheng C F 2010 Acta Phys. Sin. 59 8490 (in Chinese) [刘曼, 陈小艺, 李海霞, 宋洪胜, 滕树云, 程传福 2010 物理学报 59 8490]
[20] Pavani S R P, Greengard A, Piestun R 2009 Appl. Phys. Lett. 95 021103
[21] Blanchard M P, H Greenaway A 1999 Appl. Opt. 38 6692
[22] Li H, Yu B, Chen D N, Niu H B 2013 Acta Phys. Sin. 62 124201 (in Chinese) [李恒, 于斌, 陈丹妮, 牛憨笨 2013 物理学报 62 124201]
[23] Wu C, Chiu D T 2013 Angew. Chem. Int. Ed. 11 3086
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