Applicability of an EM-CCD for spatially resolved TIR-ICS

被引：10

作者：

Boening, Daniel ^{[1
,2
]}

Groemer, Teja W. ^{[1
,3
]}

Klingauf, Jurgen ^{[1
,2
]}

机构：

[1] Max Planck Inst Biophys Chem, D-37077 Gottingen, Germany

[2] Inst Med Phys & Biophys, Dept Cellular Biophys, D-48149 Munster, Germany

[3] Univ Erlangen Nurnberg, Dept Psychiat & Psychotherapy, D-91054 Erlangen, Germany

来源：

OPTICS EXPRESS | 2010年 / 18卷 / 13期

关键词：

FLUORESCENCE CORRELATION SPECTROSCOPY; TOTAL INTERNAL-REFLECTION; COUPLED-DEVICE CAMERA; CORRELATION MICROSCOPY; DIFFUSION; DYNAMICS; PROTEIN; CELLS;

D O I：

10.1364/OE.18.013516

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

In this work we systematically explored performance of an EM-CCD as a detector for spatially resolved total internal reflection image correlation spectroscopy (TIR-ICS) with respect to adjustable parameters. We show that variations in the observation volume (pixel binning) can be well described by a simple structural term omega. To test the sensitivity of camera-based TIR-ICS we measured diffusion coefficients and particle numbers (PN) of fluorescent probes of different sizes (Fluorospheres, GFP and labeled antibodies) at varying viscosities, concentrations, and sampling rates. TIR-ICS allowed distinguishing between different probe concentrations with differences in PN of 5% and differences of 6% in D by acquiring only 15 independent measurement runs. (C) 2010 Optical Society of America

引用

页码：13516 / 13528

页数：13

共 25 条

[1]

Bacia Kirsten, 2007, V398, P73, DOI 10.1007/978-1-59745-513-8_7

[2] Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements [J].

Dertinger, Thomas ;

Pacheco, Victor ;

von der Hocht, Iris ;

Hartmann, Rudolf ;

Gregor, Ingo ;

Enderlein, Joerg .

CHEMPHYSCHEM, 2007, 8 (03) :433-443

[3] Prionics or The kinetic basis of prion diseases [J].

Eigen, M .

BIOPHYSICAL CHEMISTRY, 1996, 63 (01) :A1-A18

[4] Total internal reflection fluorescence correlation spectroscopy (TIR-FCS) with low background and high count-rate per molecule [J].

Hassler, K ;

Leutenegger, M ;

Rigler, P ;

Rao, R ;

Rigler, R ;

Gösch, M ;

Lasser, T .

OPTICS EXPRESS, 2005, 13 (19) :7415-7423

[5] High count rates with total internal reflection fluorescence correlation spectroscopy [J].

Hassler, K ;

Anhut, T ;

Rigler, R ;

Gösch, M ;

Lasser, T .

BIOPHYSICAL JOURNAL, 2005, 88 (01) :L1-L3

[6] Fluorescence correlation spectroscopy: Novel variations of an established technique [J].

Haustein, Elke ;

Schwille, Petra .

ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 2007, 36 :151-169

[7] Spatially resolved total internal reflection fluorescence correlation microscopy using an electron multiplying charge-coupled device camera [J].

Kannan, Balakrishnan ;

Guo, Lin ;

Sudhaharan, Thankiah ;

Ahmed, Sohail ;

Maruyama, Ichiro ;

Wohland, Thorsten .

ANALYTICAL CHEMISTRY, 2007, 79 (12) :4463-4470

[8] Electron multiplying charge-coupled device camera based fluorescence correlation spectroscopy [J].

Kannan, Balakrishnan ;

Har, Jia Yi ;

Liu, Ping ;

Maruyama, Ichiro ;

Ding, Jeak Ling ;

Wohland, Thorsten .

ANALYTICAL CHEMISTRY, 2006, 78 (10) :3444-3451

[9] Fluorescence correlation spectroscopy in living cells [J].

Kim, Sally A. ;

Heinze, Katrin G. ;

Schwille, Petra .

NATURE METHODS, 2007, 4 (11) :963-973

[10] Imaging clathrin dynamics in Drosophila melanogaster hemocytes reveals a role for actin in vesicle fission [J].

Kochubey, Olexiy ;

Majumdar, Arnitabha ;

Klingauf, Jurgen .

TRAFFIC, 2006, 7 (12) :1614-1627

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