MPScope: A versatile software suite for multiphoton microscopy

被引:78
作者
Nguyen, Quoc-Thang [1 ]
Tsai, Philbert S. [1 ]
Kleinfeld, David [1 ]
机构
[1] Univ Calif San Diego, Dept Phys, La Jolla, CA 92093 USA
关键词
femtosecond laser; two-photon; electrophysiology; software; scripting; calcium imaging; histology;
D O I
10.1016/j.jneumeth.2006.03.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
MPScope is a software suite to control and analyze data from custom-built multiphoton laser scanning fluorescence microscopes. The acquisition program MPScan acquires, displays and stores movies, linescans, image stacks or arbitrary regions from up to four imaging channels and up to two analog inputs, while plotting the intensity of regions of interest in real-time. Bidirectional linescans allow 256 x 256 pixel frames to be acquired at up to 10 fps with typical galvanometric scanners. A fast stack mode combines movie acquisition with continuous z-focus motion and adjustment of laser intensity for constant image brightness. Fast stacks can be automated by custom programs running in an integrated scripting environment, allowing a 1 mm(3) cortical volume to be sampled in 1 billion voxels in approximately 1 h. The analysis program MPView allows viewing of stored frames, projections, automatic detection of cells and plotting of their average intensity across frames, direct frame transfer to Matlab, AVI movie creation and file export to ImageJ. The combination of optimized code, multithreading and COM (Common Object Model) technologies enables MPScope to fully take advantage of custom-built two-photon microscopes and to simplify their realization. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:351 / 359
页数:9
相关论文
共 20 条
[1]   2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY [J].
DENK, W ;
STRICKLER, JH ;
WEBB, WW .
SCIENCE, 1990, 248 (4951) :73-76
[2]   ANATOMICAL AND FUNCTIONAL IMAGING OF NEURONS USING 2-PHOTON LASER-SCANNING MICROSCOPY [J].
DENK, W ;
DELANEY, KR ;
GELPERIN, A ;
KLEINFELD, D ;
STROWBRIDGE, BW ;
TANK, DW ;
YUSTE, R .
JOURNAL OF NEUROSCIENCE METHODS, 1994, 54 (02) :151-162
[3]   Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons [J].
Fan, GY ;
Fujisaki, H ;
Miyawaki, A ;
Tsay, RK ;
Tsien, RY ;
Ellisman, MH .
BIOPHYSICAL JOURNAL, 1999, 76 (05) :2412-2420
[4]  
Foley J. D., 1990, Computer Graphics, Principles and Practice, V2nd
[5]   Deep tissue two-photon microscopy [J].
Helmchen, F ;
Denk, W .
NATURE METHODS, 2005, 2 (12) :932-940
[6]   Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex [J].
Kleinfeld, D ;
Mitra, PP ;
Helmchen, F ;
Denk, W .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (26) :15741-15746
[7]   Two-photon imaging in living brain slices [J].
Mainen, ZF ;
Maletic-Savatic, M ;
Shi, SH ;
Hayashi, Y ;
Malinow, R ;
Svoboda, K .
METHODS, 1999, 18 (02) :231-+
[8]   A custom-made two-photon microscope and deconvolution system [J].
Majewska, A ;
Yiu, G ;
Yuste, R .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 2000, 441 (2-3) :398-408
[9]   Targeted whole-cell recordings in the mammalian brain in vivo [J].
Margrie, TW ;
Meyer, AH ;
Caputi, A ;
Monyer, H ;
Hasan, MT ;
Schaefer, AT ;
Denk, W ;
Brecht, M .
NEURON, 2003, 39 (06) :911-918
[10]   e-Phys: a suite of intracellular neurophysiology programs integrating COM (Component Object Model) technologies [J].
Nguyen, QT ;
Miledi, R .
JOURNAL OF NEUROSCIENCE METHODS, 2003, 128 (1-2) :21-31