[1]Bertram SU,Vicky KATSOULIDOU,and Wolfgang BECKER.时间相关单光子计数记录快速荧光寿命图像[J].深圳大学学报理工版,2008,25(3):238-243.
 Bertram SU,Vicky KATSOULIDOU,and Wolfgang BECKER.Recording fast time-series of fluorescence lifetime images by TCSPC FLIM[J].Journal of Shenzhen University Science and Engineering,2008,25(3):238-243.
点击复制

时间相关单光子计数记录快速荧光寿命图像()
分享到:

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第25卷
期数:
2008年3期
页码:
238-243
栏目:
生物医学工程
出版日期:
2008-07-31

文章信息/Info

Title:
Recording fast time-series of fluorescence lifetime images by TCSPC FLIM
文章编号:
1000-2618(2008)03-0238-06
作者:
Bertram SUVicky KATSOULIDOUand Wolfgang BECKER
Becker & Hickl公司,Nahmitzer Damm 30,柏林D-12277,德国
Author(s):
Bertram SUVicky KATSOULIDOUand Wolfgang BECKER
Becker & Hickl GmbH,Nahmitzer Damm 30,Berlin D-12277,Germany
关键词:
荧光寿命成像技术时间相关单光子计数叶绿素瞬态测量
Keywords:
fluorescence lifetime imaging microscopytime-correlated single photon countingmeasurement of chlorophyll transients
分类号:
TP 271+.5;O 443;Q-3
文献标志码:
A
摘要:
基于时间相关单光子计数(TCSPC)的荧光寿命成像(FLIM)的获取时间取决于成像的图像尺寸、样品寿命的精度及样品计数率.对于高密度荧光团样品,如染色组织或植物细胞,当前可用的样品计数率与TCSPC荧光寿命成像技术最高的计数率接近.该文描述了在高计数率下TCSPC性能,并估计计数损失及堆积效应.结果表明,整个体系的寿命误差比预料的要小.因此,TCSPC FLIM可在获取时间低于1 s的情况下记录寿命图像.为增加FLIM时间序列记录,利用存储交换技术,在采集下一个光子期间读取正在记录的数据,使用两个平行TCSPC模块,可以每秒两幅图像记录荧光寿命时间序列.该技术可应用于活体植物组织叶绿素的瞬态测量.
Abstract:
The acquisition time of TCSPC FLIM depends on the number of pixels of the image,on the required lifetime accuracy,and on the count rate available from the sample.For samples with high fluorophore concentrations,such as stained tissue or plant cells the available count rates may come close to the maximum counting capability of the currently used TCSPC FLIM techniques.We describe the behaviour of TCSPC at high count rates and estimate the size of counting loss and pile-up effects.It turns out that systematic lifetime errors are smaller than previously believed.TCSPC FLIM can therefore be used to record lifetime images within acquisition times of less than 1 s.To increase the recording of FLIM time series we employed a memory swapping technique that reads out the data of one recording step during the acquisition of the next one.With parallel operation of two TCSPC modules we were able to record FLIM time series at a rate of 2 images per second.We demonstrate the technique for the measurement of chlorophyll transients in live plant tissue.

参考文献/References:

[1]Becker W.高级时间相关单光子计数技术.[M].柏林:Springer出版社,2005(英文版).
[2]Becker W,Bergmann A,Hink M A,等.利用时间相关单光子计数进行荧光寿命成像[J].显微研究技术2004,63:58-66 (英文版).
[3]Becker W,Bergmann A,Biskup C.利用时间相关单光子计数进行多光谱寿命成像[J].显微研究技术.2007,70:403-409 (英文版).
[4]Ballew R M,Demas J N.用于评估单指数衰减的一种快速寿命确定方法的误差分析[J].分析化学,1989,61(1):30-33(英文版).
[5]Govindjee.Kautsly之后的63年:叶绿素α荧光[J].澳大利亚植物生理学报,1995,22:131-160 (英文版).
[6]Biskup C,Kelbauskas L,Zimmer T,等.利用基于荧光寿命共振能量转移成像技术对PSD-95与钾离子通道相互作用进行可视化研究[J].生物医学光学学报,2004,9(4):735-759(英文版).
[7]Davey A M,Walvick R P,LIU Yue-xin,等.肥大细胞中与IgE受体交叉链接的膜有序状态和分子动力学[J].生物物理学杂志,2007,92(1):343-355.
[8]Kllner M,Wolfrum J.荧光寿命测量需要多少个光子[J].物理化学快报,1992,200(1/2):199-204.
[9]Becker W,Bergmann A,Biscotti G,等.利用时间相关单光子计数快速获取荧光寿命成像显微数据[C]// SPIE会议论文集,2004,5323:27-35(英文版).
[10]Knig K,Riemann I.利用光学层析对人类皮肤组织实现基于亚细胞和皮秒时间分辨水平的高分辨成像[J].生物医学光学学报,2003,8(3):432-439 (英文版).
[11]Becker W.Bh TCSPC使用手册[M/OL].第3版.柏林:Becker & Hickl 公司,2008.http://www.becker-hickl.com.
[12]Becker W,Hickl H,Zander C,等.利用时间相关单光子计数在微毛细管中对单分子的时间分辨探测和分类[J].科学仪器综述,1999,70(3):1835-1841(英文版).
[13]Becker & Hickl GmbH公司.DCS-120 荧光寿命成像共焦扫描系统使用手册[M].柏林:Becker & Hickl GmbH公司.http:// www.becker-hickl.com(英文版).
[14]Gerritsen H C,Asselbergs M A H,Agronskaia A V,等.扫描显微的荧光寿命成像:获取速率、光子效率和时间分辨[J].显微技术学报,2002,206(3):218224 (英文版).


[1]Becker W.Advanced Time-correlated Single-photon Counting Techniques[M].Berlin:Springer,2005.
[2]Becker W,Bergmann A,Hink M A,et al.Fluorescence lifetime imaging by time-correlated single photon counting[J].Micrsoft Research and Technique,2004,63(1):58-66.
[3]Becker W,Bergmann A,Biskup C.Multispectral fluorescence lifetime imaging by TCSPC[J].Micrsoft Research and Technique,2007,70(5):403-409.
[4]Ballew R M,Demas J N.An error analysis of the rapid lifetime determination method for the evaluation of single exponential decays[J].Analytical Chemistry,1989,61(1):30-33.
[5]Govindjee.Sixty-three years since Kautsky:Chlorophyll α fluorescence[J].Australian Journal of Plant Physiology,1995,22:131-160.
[6]Biskup C,Kelbauskas L,Zimmer T,et al.Interaction of PSD-95 with potassium channels visualized by fluorescence lifetime-based resonance energy transfer imaging[J].Journal of Biomedical Optics,2004,9(4):735-759.
[7]Davey A M,Walvick R P,LIU Yue-xin,et al.Membrane order and molecular dynamics associated with IgE receptor cross-linking in mast cells[J].Biophysical Journal,2007,92(1):343-355.
[8]Kllner M,Wolfrum J.How many photons are necessary for fluorescence-lifetime measurements[J].Chemical Physics Letters,1992,200(1/2):199-204.

[9]Becker W,Bergmann A,Biscotti G,et al.High-speed FLIM data acquisition by time-correlated single photon counting[C]// Proc SPIE,2004,5323:27-35.
[10]Knig K,Riemann I. High resolution tomography of human skin with subcellular resolution and picosecond time resolution[J].Journal of Biomedical Optics,2003,8(3):432-439.
[11]Becker W.The bh TCSPC handbook.Becker & Hickl GmbH[M/OL].3rd edition. http://www.becker-hickl.com,2008.
[12]Becker W,Hickl H,Zander C,et al.Time-resolved detection and identification of single analyte molecules in microcapillaries by time-correlated single photon counting[J].Review of Scientific Instruments,1999,70(3):1835-1841.
[13]Becker & Hickl GmbH.DCS-120 confocal scanning FLIM systems,user handbook[M/OL].Berlin:Becker & Hickl GmbH.http://www.becker-hickl.com.
[14]Gerritsen H C,Asselbergs M A H,Agronskaia A V,et al.Fluorescence lifetime imaging in scanning microscopes:acquisition speed,photon economy and lifetime resolution[J].Journal of Microscopy,2002,206(3):218224.

相似文献/References:

[1]Wolfgang BECKER.多维时间相关单光子计数在生物医学中的应用[J].深圳大学学报理工版,2008,25(3):221.
 Wolfgang BECKER.Multi-dimensional time-correlated single photon counting:a fast and efficient optical recording technique for biomedical spectroscopy[J].Journal of Shenzhen University Science and Engineering,2008,25(3):221.
[2]程颖,ABDULLA S.,CHORVAT D. J. R.,等.基于多维TCSPC技术的心肌细胞Ca2+动力学研究[J].深圳大学学报理工版,2008,25(4):369.
 CHENG Ying,ABDULLA S.,CHORVAT D. J. R.,et al.Investigation of intracellular calcium dynamics during cardiac cell contraction by multi-dimensional TCSPC[J].Journal of Shenzhen University Science and Engineering,2008,25(3):369.

备注/Memo

备注/Memo:
收稿日期:2008-05-29
作者简介:Bertram SU(1976-),男(德国人),德国Becker & Hickl公司博士.E-mail:bsu@becker-hickl.com
*本文为2008年第3届国际生物医学光学方法高级研讨会宣读论文
更新日期/Last Update: 2008-08-07