Mapping microscope object polarized emission to the back focal plane pattern.
Mapping microscope object polarized emission to the back focal plane pattern.
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DOI：
10.1117/1.3155520
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发表时间：
2009-05
影响因子：
3.5
通讯作者：
Ajtai K
中科院分区：
文献类型：
--
作者：
Burghardt TP;Ajtai K
The back focal plane (BFP) intensity pattern from a high-aperture objective separately maps far- and near-field emission from dipoles near a bare glass or metal-film-coated glass/aqueous interface. Total internal reflection (TIR) excitation of a fluorescent sample gave a BFP pattern interpreted in terms of fluorescent dipole orientation and distance from the interface. Theoretical consideration of this system led to identification of emission characteristics that remove a dipole orientation degeneracy in conventional microscope fluorescence polarization measurements. BFP pattern inspection removes the degeneracy. Alternatively, a BFP mask blocking a small fraction of emitted light in a standard imaging microscope prevents uniform collection of the BFP intensity and also eliminates the degeneracy. The BFP pattern from a single photoactivated photoactivatable green fluorescent protein (PAGFP) tagged myosin in a muscle fiber was observed despite the large background light from the highly concentrated myosin tagged with unphotoactivated PAGFP. This was accomplished by imaging the pattern from a nontelecentric plane, where most of the background intensity’s pattern was translated laterally from the single-molecule object’s pattern. TIR/BFP pattern imaging requires a simple alteration of the fluorescence microscope and is consistent with single-molecule imaging in a fluorophore dense three-dimensional object like a muscle fiber.
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DOI：
10.1073/pnas.0507134103
发表时间：
2006-04-25
影响因子：
11.1
作者：
Toprak, E;Enderlein, J;Selvin, PR
通讯作者：
Selvin, PR
DOI：
10.1364/josab.4.000337
发表时间：
1987-03-01
影响因子：
1.9
作者：
HELLEN, EH;AXELROD, D
通讯作者：
AXELROD, D
影响因子：
3.3
作者：
Tikunov, BA;Sweeney, HL;Rome, LC
通讯作者：
Rome, LC
影响因子：
3.4
作者：
Borejdo, J.;Gryczynski, Z.;Gryczynski, I.
通讯作者：
Gryczynski, I.
影响因子：
2.7
作者：
Fulbright, R M;Axelrod, D
通讯作者：
Axelrod, D