Dual-color fluorescence cross-correlation spectroscopy on a planar optofluidic chip.

Fluorescence cross-correlation spectroscopy (FCCS) is a highly sensitive fluorescence technique with distinct advantages in many bioanalytical applications involving interaction and binding of multiple components. Due to the use of multiple beams, bulk optical FCCS setups require delicate and complex alignment procedures. We demonstrate the first implementation of dual-color FCCS on a planar, integrated optofluidic chip based on liquid-core waveguides that can guide liquid and light simultaneously. In this configuration, the excitation beams are delivered in predefined locations and automatically aligned within the excitation waveguides. We implement two canonical applications of FCCS in the optofluidic lab-on-chip environment: particle colocalization and binding/dissociation dynamics. Colocalization is demonstrated in the detection and discrimination of single-color and double-color fluorescently labeled nanobeads. FCCS in combination with fluorescence resonance energy transfer (FRET) is used to detect the denaturation process of double-stranded DNA at nanomolar concentration.

Ultralow power trapping and fluorescence detection of single particles on an optofluidic chip.

The development of on-chip methods to manipulate particles is receiving rapidly increasing attention. All-optical traps offer numerous advantages, but are plagued by large required power levels on the order of hundreds of milliwatts and the inability to act exclusively on individual particles. Here, we demonstrate a fully integrated electro-optical trap for single particles with optical excitation power levels that are five orders of magnitude lower than in conventional optical force traps. The trap is based on spatio-temporal light modulation that is implemented using networks of antiresonant reflecting optical waveguides. We demonstrate the combination of on-chip trapping and fluorescence detection of single microorganisms by studying the photobleaching dynamics of stained DNA in E. coli bacteria. The favorable size scaling facilitates the trapping of single nanoparticles on integrated optofluidic chips.

Optofluidic particle concentration by a long-range dual-beam trap.

Ultrahigh sensitivity detection of particles in solution implies the ability to detect at very low concentrations. At the single-particle level, this is achieved through fluorescence detection, reaching down to single fluorophores. Sensitivity may also be improved by concentrating many particles into a compact cluster, thus "integrating" the signal of many particles. We show how both ways can be combined on an optofluidic chip in a fully planar geometry utilizing counterpropagating liquid-core waveguide modes to form a loss-based optical trap. This all-optical concentrator can increase the concentration of particles by more than 2 orders of magnitude and also provides a convenient, nondispersive means of transport for particle ensembles.

Loss-based optical trap for on-chip particle analysis.

Optical traps have become widespread tools for studying biological objects on the micro and nanoscale. However, conventional laser tweezers and traps rely on bulk optics and are not compatible with current trends in optofluidic miniaturization. Here, we report a new type of particle trap that relies on propagation loss in confined modes in liquid-core optical waveguides to trap particles. Using silica beads and E. coli bacteria, we demonstrate unique key capabilities of this trap. These include single particle trapping with micron-scale accuracy at arbitrary positions over waveguide lengths of several millimeters, definition of multiple independent particle traps in a single waveguide, and combination of optical trapping with single particle fluorescence analysis. The exclusive use of a two-dimensional network of planar waveguides strongly reduces experimental complexity and defines a new paradigm for on-chip particle control and analysis.

Ultrasensitive Qbeta phage analysis using fluorescence correlation spectroscopy on an optofluidic chip.

We demonstrate detection and analysis of the Qbeta bacteriophage on the single virus level using an integrated optofluidic biosensor. Individual Qbeta phages with masses on the order of attograms were sensed and analyzed on a silicon chip in their natural liquid environment without the need for virus immobilization. The diffusion coefficient of the viruses was extracted from the fluorescence signal by means of fluorescence correlation spectroscopy (FCS) and found to be 15.90+/-1.50 microm(2)/s in excellent agreement with previously published values. The aggregation and disintegration of the phage were also observed. Virus flow velocities determined by FCS were in the 60-300 microm/s range. This study suggests considerable potential for an inexpensive and portable sensor capable of discrimination between viruses of different sizes.

Microphotonic control of single molecule fluorescence correlation spectroscopy using planar optofluidics.

We demonstrate the implementation of fluorescence correlation spectroscopy (FCS) on a chip. Full planar integration is achieved by lithographic definition of sub-picoliter excitation volumes using intersecting solid and liquid-core optical waveguides. Concentration dependent measurements on dye molecules with single molecule resolution are demonstrated. Theoretical modeling of the FCS autocorrelation function in microstructured geometries shows that the FCS behavior can be controlled over a wide range by tailoring the micro-photonic environment. The ability to perform correlation spectroscopy using silicon photonics without the need for free-space microscopy permits implementation of numerous diagnostic applications on compact planar optofluidic devices.