In Vivo Fluorescence Correlation and Cross-Correlation Spectroscopy

被引：1

作者：

Muetze, Joerg ^{[1
]}

Ohrt, Thomas ^{[1
]}

Petrasek, Zdenek ^{[1
]}

Schwille, Petra ^{[1
]}

机构：

[1] Tech Univ Dresden, Biophys Grp, Biotechnol Zentrum, D-01307 Dresden, Germany

来源：

SINGLE MOLECULE SPECTROSCOPY IN CHEMISTRY, PHYSICS AND BIOLOGY | 2010年 / 96卷

关键词：

RESONANCE ENERGY-TRANSFER; ASYMMETRIC CELL-DIVISION; LIVING CELLS; C-ELEGANS; 2-PHOTON EXCITATION; SINGLE MOLECULES; PROTEIN DYNAMICS; PAR PROTEINS; DIFFUSION; FLUCTUATIONS;

D O I：

10.1007/978-3-642-02597-6_7

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

In this manuscript, we describe the application of Fluorescence Correlation Spectroscopy (MS), Fluorescence Cross-Correlation Spectroscopy (FCCS), and scanning FCS (sFCS) to two in vivo systems. In the first part, we describe the application of two-photon standard and scanning FCS in Caenorhabditis elegans embryos. The differentiation of a single fertilized egg into a complex organism in C. elegans is regulated by a number of protein-dependent processes. The oocyte divides asymmetrically into two daughter cells of different developmental fate. Two of the involved proteins, PAR-2 and NMY-2, are studied. The second investigated system is the mechanism of RNA interference in human cells. An EGFP based cell line that allows to study the dynamics and localization of the RNA-induced silencing complex (RISC) with FCS in vivo is created, which has so far been inaccessible with other experimental methods. Furthermore, Fluorescence Cross-Correlation Spectroscopy is employed to highlight the asymmetric incorporation of labeled siRNAs into RISC.

引用

页码：139 / 154

页数：16

共 38 条

[1] Fluorescence cross-correlation spectroscopy in living cells [J].

Bacia, K ;

Kim, SA ;

Schwille, P .

NATURE METHODS, 2006, 3 (02) :83-89

[2] Scanning two-photon fluctuation correlation spectroscopy: Particle counting measurements for detection of molecular aggregation [J].

Berland, KM ;

So, PTC ;

Chen, Y ;

Mantulin, WW ;

Gratton, E .

BIOPHYSICAL JOURNAL, 1996, 71 (01) :410-420

[3] C-elegans PAR proteins function by mobilizing and stabilizing asymmetrically localized protein complexes [J].

Cheeks, RJ ;

Canman, JC ;

Gabriel, WN ;

Meyer, N ;

Strome, S ;

Goldstein, B .

CURRENT BIOLOGY, 2004, 14 (10) :851-862

[4] Molecular brightness characterization of EGFP in vivo by fluorescence fluctuation spectroscopy [J].

Chen, Y ;

Müller, JD ;

Ruan, QQ ;

Gratton, E .

BIOPHYSICAL JOURNAL, 2002, 82 (01) :133-144

[5]

Chendrimada TP, 2007, NATURE, V447, P823, DOI 10.1038/nature05841

[6] Acto-myosin reorganization and PAR polarity in C-elegans [J].

Cowan, Carrie R. ;

Hyman, Anthony A. .

DEVELOPMENT, 2007, 134 (06) :1035-1043

[7] Asymmetric cell division in C-elegans:: Cortical polarity and spindle positioning [J].

Cowan, CR ;

Hyman, AA .

ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY, 2004, 20 :427-453

[8] Single-pair fluorescence resonance energy transfer on freely diffusing molecules: Observation of Forster distance dependence and subpopulations [J].

Deniz, AA ;

Dahan, M ;

Grunwell, JR ;

Ha, TJ ;

Faulhaber, AE ;

Chemla, DS ;

Weiss, S ;

Schultz, PG .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (07) :3670-3675

[9] 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

[10] Measuring fast dynamics in solutions and cells with a laser scanning microscope [J].

Digman, MA ;

Brown, CM ;

Sengupta, P ;

Wiseman, PW ;

Horwitz, AR ;

Gratton, E .

BIOPHYSICAL JOURNAL, 2005, 89 (02) :1317-1327

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