A Fluorescence Detection and Selection Device for Dna-Molecules Bound to Microspheres

Holm, Johan; Elderstig, H.; Kristensen, Ove; Rigler, Rudolf

doi:10.1080/07328319708002916

Cited by 10 publications

(7 citation statements)

References 5 publications

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“…To obtain a transparent detection window within the capillary, the outer polyacrylamide coating is removed by a flame. All microstructures used in the experiments are provided either by the IMC, Kista, Sweden or M. Stuke, Laser processing, MPI for Biophysical Chemistry, Am Fassberg, D-37077 Göttingen, Germany. ,− The microstructures are filled by capillary forces or a slight vacuum (200 mbar) was applied.…”

Section: Methodsmentioning

confidence: 99%

Two-Beam Cross-Correlation: A Method To Characterize Transport Phenomena in Micrometer-Sized Structures

et al. 1999

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To determine flow properties, namely, the velocity and angle of the flow in microstructured channels, an experimental realization based on fluorescence correlation spectroscopy is described. For this purpose, two micrometer-sized spatially separated volume elements have been created. The cross-correlation signal from these has been recorded and evaluated mathematically. In addition to previous results, two-beam cross-correlation allows for fast and easy determination of even small (down to 200 μm/s) flow velocities, as well as simultaneous measurement of diffusion properties of single dye molecules within a rather short detection time of 5-100 s and an error rate of less than 20%. The spatial flow resolution is around 1-2 μm, limited by the diameter of the volume element. Furthermore, vectorial flow data can be obtained and evaluated. A discussion of the theoretical background and an experimental verification of the theoretical results is performed. The feasibility of fast and easy data processing is shown if the flow time is the only desired information. Possible applications of this precise and simple method are the determination of transportation effects within artificial microstructures for CE and HPLC, fast chemical kinetics, and high-throughput screening.

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Section: Methodsmentioning

confidence: 99%

Two-Beam Cross-Correlation: A Method To Characterize Transport Phenomena in Micrometer-Sized Structures

et al. 1999

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“…The fabrication method requires only one mask layer and few processing steps and makes it possible to produce channels close to the surface of the wafer which facilitates liquid handling in many chemical analysis systems, including attachments of various fluid-transfer fittings. 38,39…”

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confidence: 99%

Electroosmosis- and Pressure-Driven Chromatography in Chips Using Continuous Beds

et al. 1999

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The application range of microchips can be extended to any mode of chromatography by filling the narrow channels with continuous polymer beds, exemplified by electrochromatography and ion-exchange chromatography. “Wall effects” are eliminated by anchoring the bed to the wall of the channel, an arrangement which has the additional advantage that no frits to support the bed are required. The design of the equipment is based on a quartz chip with all auxiliary pieces (for example, electrode vessels and fluid transfer fittings) placed in a rack, which permits a flexibility of great importance for automation. The same resolution and van Deemter plots were obtained in experiments performed in fused-silica capillaries and in chips for both low-molecular-weight (alkyl phenones, antidepressants) and high-molecular-weight substances (proteins). A sample of uracil, phenol, and benzyl alcohol was separated by electrochromatography in less than 20 s.

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“…In this case, electroosmotic flow would be of advantage, because the flow direction can be changed almost instantaneously. 24 A flow velocity of several millimeters per second is desirable for high-throughput screening. 25 The hydrodynamic flow is, for such experiments, a simple way to induce high flow velocity and avoid high voltages at the same time.…”

Section: Resultsmentioning

confidence: 99%

“…This step is time-consuming and leads to irregularities in the laminar flow. In this case, electroosmotic flow would be of advantage, because the flow direction can be changed almost instantaneously …”

Section: Resultsmentioning

confidence: 99%

Hydrodynamic Flow Profiling in Microchannel Structures by Single Molecule Fluorescence Correlation Spectroscopy

et al. 2000

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In this paper we demonstrate high spatial resolution hydrodynamic flow profiling in silicon wafer based microchannels using single molecule fluorescence correlation spectroscopy (FCS). We have used confocal fluorescence microscopy to detect single tetramethylrhodamine (TMR-4-dUTP) biomolecules traversing a approximately 1 fL volume element defined by an argon laser beam focus. By elevating a (approximately 10(-10) M) reservoir of diluted analyte, a continuous hydrodynamic flow through the microstructure could be accomplished. The microchannel was then scanned with a diffraction-limited focus in approximately 1-microm steps in both the vertical and the horizontal directions to determine the flow profile across a 50 x 50 microm2 channel. The flow profile measured was parabolic in both dimensions, thereby showing a Poiseuille laminar flow profile. Future microstructures can hereby be nondestructively investigated with the use of high spatial resolution confocal correlation microscopy.

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A Fluorescence Detection and Selection Device for Dna-Molecules Bound to Microspheres

Cited by 10 publications

References 5 publications

Two-Beam Cross-Correlation: A Method To Characterize Transport Phenomena in Micrometer-Sized Structures

Two-Beam Cross-Correlation: A Method To Characterize Transport Phenomena in Micrometer-Sized Structures

Electroosmosis- and Pressure-Driven Chromatography in Chips Using Continuous Beds

Hydrodynamic Flow Profiling in Microchannel Structures by Single Molecule Fluorescence Correlation Spectroscopy

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