The role of photon statistics in fluorescence anisotropy imaging

被引：54

作者：

Lidke, KA ^{[1
]}

Rieger, B ^{[1
]}

Lidke, DS ^{[1
]}

Jovin, TM ^{[1
]}

机构：

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

来源：

IEEE TRANSACTIONS ON IMAGE PROCESSING | 2005年 / 14卷 / 09期

关键词：

confocal microscopy; energy migration fluorescence resonance energy transfer (emFRET); enhanced green fluorescent protein (eGFP); scale selective filtering;

D O I：

10.1109/TIP.2005.852458

中图分类号：

TP18 [人工智能理论];

学科分类号：

081104 ; 0812 ; 0835 ; 1405 ;

摘要：

Anisotropy imaging can be used to image resonance energy transfer between pairs of identical fluorophores and, thus, constitutes a powerful tool for monitoring protein homo-association in living single cells. The requirement for only a single fluorophore significantly simplifies biological preparation and interpretation. We use quantitative methods for the acquisition and image processing of anisotropy data that return the expected error of the anisotropy per pixel based on photon statistics. The analysis methods include calibration procedures and allow for a balance in spatial, anisotropy, and temporal resolution. They are featured here with anisotropy images of fluorescent calibration beads and enhanced green fluorescent protein complexes in live cells.

引用

页码：1237 / 1245

页数：9

共 28 条

[1]

AXELROD D, 1989, METHOD CELL BIOL, V30, P333

[2] Depolarization by high aperture focusing [J].

Bahlmann, K ;

Hell, SW .

APPLIED PHYSICS LETTERS, 2000, 77 (05) :612-614

[3]

BAKKER P, 1999, P C COMP VIS PATT RE, V2, P535

[4] Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM) [J].

Clayton, AHA ;

Hanley, QS ;

Arndt-Jovin, DJ ;

Subramaniam, V ;

Jovin, TM .

BIOPHYSICAL JOURNAL, 2002, 83 (03) :1631-1649

[5] Homo-FRET microscopy in living cells to measure monomer-dimer transition of GFP-tagged proteins [J].

Gautier, I ;

Tramier, M ;

Durieux, C ;

Coppey, J ;

Pansu, RB ;

Nicolas, JC ;

Kemnitz, K ;

Coppey-Moisan, M .

BIOPHYSICAL JOURNAL, 2001, 80 (06) :3000-3008

[6] Ensemble and single particle photophysical proper-ties (Two-Photon excitation, anisotropy, FRET, lifetime, spectral conversion) of commercial quantum dots in solution and in live cells [J].

Grecco, HE ;

Lidke, KA ;

Heintzmann, R ;

Lidke, DS ;

Spagnuolo, C ;

Martinez, OE ;

Jares-Erijman, EA ;

Jovin, TM .

MICROSCOPY RESEARCH AND TECHNIQUE, 2004, 65 (4-5) :169-179

[7] 3-PHOTON INDUCED FLUORESCENCE OF 2,5-DIPHENYLOXAZOLE WITH A FEMTOSECOND TI-SAPPHIRE LASER [J].

GRYCZYNSKI, I ;

MALAK, H ;

LAKOWICZ, JR .

CHEMICAL PHYSICS LETTERS, 1995, 245 (01) :30-35

[8]

Hendriks C.L.L., 1999, DIPIMAGE SCI IMAGE P

[9] Measurement of the Detective Quantum Efficiency (DQE) of digital X-ray imaging devices according to the standard IEC 62220-1 [J].

Illers, H ;

Buhr, E ;

Bergmann, D ;

Hoeschen, C .

MEDICAL IMAGING 2004: PHYSICS OF MEDICAL IMAGING, PTS 1 AND 2, 2004, 5368 :177-187

[10] FRET imaging [J].

Jares-Erijman, EA ;

Jovin, TM .

NATURE BIOTECHNOLOGY, 2003, 21 (11) :1387-1395

← 1 2 3 →