Detection of single molecules of phycoerythrin in hydrodynamically focused flows by laser-induced fluorescence

Nguyen, Dinh C.; Keller, Richard A.; Jett, James H.; Martin, John

doi:10.1021/ac00144a032

Cited by 218 publications

(105 citation statements)

References 12 publications

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“…Although SMD of molecules with numerous (-30) chromophores has been possible for some time (10)(11)(12) To achieve further gains in SMD, we have reduced the optical excitation volume even further-indeed, to subwavelength proportions in all three dimensions. This is accomplished using illumination mode near-field scanning optical microscopy (NSOM) (13) (15).…”

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confidence: 99%

Single Molecules Observed by Near-Field Scanning Optical Microscopy

Betzig

Chichester

1993

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1,310

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Individual carbocyanine dye molecules in a sub-monolayer spread have been imaged with near-field scanning optical microscopy. Molecules can be repeatedly detected and spatially localized (to -X/50 where X is the wavelength of light) with a sensitivity of at least 0.005 molecules/(Hz)"12 and the orientation of each molecular dipole can be determined. This information is exploited to map the electric field distribution in the near-field aperture with molecular spatial resolution. Fig. iD) is consistent with the emission expected from a single diIC12(3) molecule under the known excitation conditions (21). Fourth, each structure in Fig. 1 has a well-defined dipole orientation as discussed below. Finally,

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confidence: 99%

Single Molecules Observed by Near-Field Scanning Optical Microscopy

Betzig

Chichester

1993

Science

1,310

830

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“…Laser-induced fluorescence has been a very sensitive analytical technique 1 , and one-molecule detection [2][3][4] has been achieved. It has progressed so well that the greatest concern for a highly sensitive determination is not the increase in the signal but the decrease in laser light scattering.…”

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Single Molecule Detection by Laser Two-Photon Excited Fluorescence in a Capillary Flowing Cell

et al. 1998

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The two-photon excited fluorescence of fluorescein and Rhodamine B in a flowing sample cell was observed using a self mode-locked Ti-sapphire laser (180 fs pulse). The detection limit of fluorescein was less than 2 molecules in the probe volume. The single molecule detection of Rhodamine was successfully performed in a flowing cell, and the number of photon bursts agreed approximately with that expected.

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“…PE conjugates are also widely used as fluorescence markers for cell sorting (60), so an understanding of their biosynthesis could have biotechnological implications. The fluorescence quantum yield for the CpeY/CpeZ-generated CpeA was 0.72 and for the CpeSgenerated CpeB-PEB was 0.89 (Table 1); most PE subunits (which may have two or three bilins attached) have quantum yields ranging between 0.84 and 0.98 (61), which makes them excellent fluorescent markers.…”

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confidence: 99%