High-speed 2D and 3D fluorescence microscopy of cardiac myocytes

被引:54
作者
Kumar, Sunil [1 ]
Wilding, Dean [1 ]
Sikkel, Markus B. [2 ]
Lyon, Alexander R. [2 ]
MacLeod, Ken T. [2 ]
Dunsby, Chris [1 ,3 ]
机构
[1] Univ London Imperial Coll Sci Technol & Med, Dept Phys, Photon Grp, Blackett Lab, London SW7 2AZ, England
[2] Univ London Imperial Coll Sci Technol & Med, Natl Heart & Lung Inst, London SW3 6LY, England
[3] Univ London Imperial Coll Sci Technol & Med, Dept Med, London W12 0NN, England
基金
英国工程与自然科学研究理事会;
关键词
PLANE ILLUMINATION MICROSCOPY; CA2+ SPARKS; VENTRICULAR MYOCYTES; EXCITATION; WAVES; HEART;
D O I
10.1364/OE.19.013839
中图分类号
O43 [光学];
学科分类号
070207 ; 0803 ;
摘要
Oblique plane microscopy (OPM) is a light sheet microscopy technique that uses a single high numerical aperture microscope objective to both illuminate a tilted plane within the specimen and to obtain an image of the tilted illuminated plane. In this paper, we present a new OPM configuration that enables both the illumination and detection focal planes to be swept simultaneously and remotely through the sample volume, enabling high speed volumetric imaging. We demonstrate the high speed imaging capabilities of the system by imaging calcium dynamics in cardiac myocytes in 2D at 926 frames per second and in 3D at 21 volumes per second. In the future, higher frame rate CCD cameras will enable volumetric imaging at much greater rates, leading to new capabilities to study dynamic events in cells at high speeds in two and three dimensions. (C) 2011 Optical Society of America
引用
收藏
页码:13839 / 13847
页数:9
相关论文
共 23 条
[1]   An optical technique for remote focusing in microscopy [J].
Botcherby, E. J. ;
Juskaitis, R. ;
Booth, M. J. ;
Wilson, T. .
OPTICS COMMUNICATIONS, 2008, 281 (04) :880-887
[2]   Aberration-free optical refocusing in high numerical aperture microscopy [J].
Botcherby, Edward J. ;
Juskaitis, Rimas ;
Booth, Martin J. ;
Wilson, Tony .
OPTICS LETTERS, 2007, 32 (14) :2007-2009
[3]   Real-time slit scanning microscopy in the meridional plane [J].
Botcherby, Edward J. ;
Booth, Martin J. ;
Juskaitis, Rimas ;
Wilson, Tony .
OPTICS LETTERS, 2009, 34 (10) :1504-1506
[4]   Quarky Calcium Release in the Heart [J].
Brochet, Didier X. P. ;
Xie, Wenjun ;
Yang, Dongmei ;
Cheng, Heping ;
Lederer, W. Jonathan .
CIRCULATION RESEARCH, 2011, 108 (02) :210-218
[5]  
Cheng H, 1996, AM J PHYSIOL-CELL PH, V270, pC148
[6]   Excitation-contraction coupling in heart: New insights from Ca2+ sparks [J].
Cheng, H ;
Lederer, MR ;
Xiao, RP ;
Gomez, AM ;
Zhou, YY ;
Ziman, B ;
Spurgeon, H ;
Lakatta, EG ;
Lederer, WJ .
CELL CALCIUM, 1996, 20 (02) :129-140
[7]   Optically sectioned imaging by oblique plane microscopy [J].
Dunsby, C. .
OPTICS EXPRESS, 2008, 16 (25) :20306-20316
[8]   Fast three-dimensional fluorescence imaging of activity in neural populations by objective-coupled planar illumination microscopy [J].
Holekamp, Terrence F. ;
Turaga, Diwakar ;
Holy, Timothy E. .
NEURON, 2008, 57 (05) :661-672
[9]   Optical sectioning deep inside live embryos by selective plane illumination microscopy [J].
Huisken, J ;
Swoger, J ;
Del Bene, F ;
Wittbrodt, J ;
Stelzer, EHK .
SCIENCE, 2004, 305 (5686) :1007-1009
[10]   Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM) [J].
Huisken, Jan ;
Stainier, Didier Y. R. .
OPTICS LETTERS, 2007, 32 (17) :2608-2610