Invented by Mark M. Wang, Eugene Tu, James P. O’Connell, Kristie L. Lykstad, William F. Butler, Biora Therapeutics Inc
The Biora Therapeutics Inc invention works as follows
Apparatus, and methods, are provided to interact light with particles in a unique and highly beneficial manner, including biological matter, such as cells. Optophoresis involves exposing particles to different optical forces. This includes optical gradient forces and moving optical gradients. This technology is a useful way to probe the inner workings a cell without using any dyes or labels. “In one aspect, it is possible to separate particles by illuminating a sorting area with a moving gradient and then allowing the particles to flow laminarly between the input and output of a first restricted path. The second constrained pathway also includes an output.
Background for Method of separation of particles
Separation and characterization of particle has many applications from industrial to biological to environmental. In the field of biotechnology and medicine, separation of cells is a common application. Sorting technologies have traditionally focused on physical characteristics such as particle size and density or affinity interactions such as receptor-ligand interaction or reactions with immune targets.
The electromagnetic response properties of the materials were used for particle sorting. Dielectrophoretic Separators, for example, use non-uniform DC and AC electric fields to separate particles. See, e.g., U.S. Pat. No. 5,814,200, Pethig et al., entitled ?Apparatus for Separating By Dielectrophoresis?. Dielectrophoresis has been used to sort cells. In Becker et. al. (with Gascoyne), PNAS USA Vol. 92, pp. In the article entitled “Separation Of Human Breast Cancer Cells From Blood By Differential Dialectic Affinity”, published in Cell Biology (January 1995), the authors stated that the dielectric properties were sufficiently different between the diseased and normal cells to allow separation. The system balanced the hydrodynamic and dielectrophoretic force acting on cells in a dielectric column containing microelectrodes. Separation systems of greater sophistication have been developed. See, e.g., Cheng, et al., U.S. Pat. No. No. 6,071,394, “Channelless Separation of Bioparticles On a Bioelectronic chip by Dielectrophoresis” Others have tried to separate particles using electrostatic forces. See, e.g., Judy et al., U.S. Pat. No. No. 4,440,638, entitled ‘Surface Field Effect Device for Manipulation of Charged Species’, and Washizu -?Electrostatic manipulation of biological objects?, Journal of Electrostatics Vol. 25, No. 1, June 1990, pp. 109-103.
Light is used to trap and sort particles. Arthur Ashkin, at Bell Laboratories was one of the first workers to use a laser. He used it for manipulating transparent?m-sized latex beads. Ashkin’s U.S. Pat. No. No. 3,808,550 entitled “Apparatuses For Trapping and AcceleratingNeutral Particles” The patent disclosed systems to trap or contain particles by radiation pressure. Lasers that generate coherent optical radiation are the most preferred sources of optical pressure. Ashkin Bell Labs researchers, Steven Chu included, were awarded the Nobel Prize for their work using optical radiation to trap particles. Chu, S. “Laser trapping of Neutral particles?,” Sci. Am., February 1992, p.71; Chu, S. “Laser Manipulation Atoms and Particles”, Science 253, pages 861-866 (1991). 861-866 (1991).
The dielectric constant is B, and r is a particle’s radius. The dielectric constant is equal to the particle’s radius, I is its light intensity measured in watts/square centimeter. The spatial derivative is shown in FIG. FIG. FIG. The optical tweezer is a highly concentrated beam that is directed at the particle.
As shown in FIG. The focused beam 12 first condenses on the particle 10, and then diverges. The intensity pattern 14 is the cross section of the beam’s intensity in the horizontal direction, while the intensity pattern 16, the cross section of intensity in vertical direction. The equation shows that the force of trapping is dependent on the gradient in the intensity of light. The force is greatest where the intensity of the light changes rapidly and is lowest where it is uniform.
Early-stable optical traps levitated particle with a horizontal laser beam, balancing upward scattering force and downward gravitational forces. The particle was kept on the optical axis by the gradient force of light. Ashkin, “Optical Levitation By Radiation Pressure”, Appl. Phys. Lett., 19(6), pp. 283-285 (1971). Ashkin revealed a trap in 1986 based on a laser beam that is highly focused, rather than light that propagates along an axis. The laser beam is so focused that it produces a tiny point of space with an extremely high intensity. Extreme focusing creates a gradient force that pulls the dielectric particle towards this point. In certain conditions, the gradient forces overcomes the scattering forces, which would otherwise force the particle out of the focal area in the direction the light. To achieve such a high degree of focusing, a laser beam is typically directed through a microscope objective with a large numerical aperture. This arrangement increases the relative contribution of high numerical aperture illumination, but reduces the effect from scattering forces.
In 1987 Ashkin reported on an experimental demonstration of optical manipulation and trapping of biological materials using a single-beam gradient force optical system. Ashkin et. al., ‘Optical Trapping of Bacteria and Viruses?, Science 20 March 1987, Vol. 235, No. 4795, pp. 1517-1520. No. 1517-1520. No. No. 4,893,886, Ashkin and al., entitled “Non-Destructive Optic Trap for Bioparticles and Method to Do Same”, reported the successful trapping biological particles using an infrared laser light source. It was claimed that the use of an infrared light source emitting coherent infrared light, whose wavelengths were stated to be between 0.8 m and 1.8 m, allowed biological materials to continue reproducing even after several life cycles of trapping at a laser power 160 mW. The term “opticution” is used. The term “optic radiation” has been used in the industry to describe optical radiation that kills biological materials.
Researchers have used light to study biological materials. The manipulation of plant cells from the inside has been demonstrated. Ashkin, et al., PNAS USA, Vol. 86, 7914-7918 (1989). Ashkin, A.: Optical Trapping of Neutral Particles and Manipulation Using Lasers, PNAS USA Vol. 94, pp. Physics, May 1997. 4853-4860. Different mechanical and force measurements were made, including the measurement of torsional compliant of bacterial flagellas by twisting a bacteria about a flagellum tethered. Block, S., et al., Nature (London), 338, pp. 514-518 (1989). The micromanipulation and manipulation of particles was demonstrated. The use of optical tweezers, in conjunction with a microbeam laser cutting technique, also known as laser scissors, or scalpel for cutting moving organelles and cells was demonstrated. Seeger, et al., Cytometry, 12, pp. 497-504 (1991). In all-optical in Vitro Fertilization, optical tweezers or scissors were used. Tadir, Y. Human Reproduction, 6 pp. 1011-1016 (1991). Handles have been used in a variety of techniques. A structure is attached to the biological material in order to assist with trapping. Block, Nature, London, 348, pp. 348-352 (1990).
Various measurements of biological systems have been made using optical trapping and position monitoring interferometric with subnanometer precision. Svoboda, Nature (London), 365, pp. 721-727 (1993). Others have proposed feedback-based systems that use a tweezer. Molloy, et al., Biophys. J., 68, pp. 2985-3055 (1995).
A number workers have attempted to distort and stretch biological materials. Ashkin, Nature (London), 332 pp. 769-771 (1987), optical tweezers were used to distort red blood cells’ shape. Multiple optical tweezers were used to create an assay for measuring the shape recovery of red blood cells. Bronkhorst, Biophys. J., 69, pp. 1666-1673 (1995). Kas, et. al. have proposed an “optical stretcher” In U.S. Pat. No. No. The system can be used to detect single malignant cells of cancer. Another assay involves colliding two particles or cells under controlled conditions. This is called the OPTCOL, for optical collision. Mammer’s Chem & Biol. 3, pp. 757,763 (1996).
Others have suggested using optical forces to measure an object’s property. See, for example, Guanming Lai and others, “Determination Of Spring Constant Of Laser-TrappedParticle By Self-Mining Interferometry”, Proc. See, for example, Guanming, Lai et al., “Determination of Spring Constant of Laser-Trapped Particle by Self-Mining Interferometry”, Proc. 197-204 (2000). Others have used the optical trapping forces balanced against fluidic drag forces as a way to calibrate an optical trap. These systems rely heavily on drag forces, especially Stokes drag forces.
Others have used light intensity patterns to position materials.” Patent No. No. In U.S. Patent No. 5,245,466, Burnes and al. entitled “Optical Matter”, arrays of extended non-crystalline and crystalline structures can be created by using light beams coupled with microscopic, polarizable material. The polarizable material adopts a pattern of applied, patterned distribution of light intensity. New Scientist No. 1889, November 18, 1989: ‘Matter Rides On Ripples of Lights’, reporting Burns’ work. 1691. Others have also proposed methods to deposit atoms onto a substrate using a standing-wave optical pattern. By translating the standing-wave pattern, it is possible to create an array of different structures. See, Celotta et al., U.S. Pat. No. No.
Yet other have tried to cause particle motion by using light. Brian Space, et. al. used a technique called ‘photophoresis’ to use a polarized light beam to cause rotary motion in the molecules. The goal was to create a gradient of concentration of molecules. The technique is best suited for propeller-shaped molecules. This allows the rotary motion to result in translation.
Various attempts to create microfluidic system have been made, used for various purposes such as sample preparation or sorting. See, e.g., Ramsey, U.S. Pat. No. No. 6,033,546, entitled “Apparatus for performing microfluidic manipulations for chemical analysis and synthesis”. Many companies, including Aclara, Caliper and others, are trying to create microsystems that include a “lab on a chips”.
Others have tried to combine optical systems with microfabricated devices. Chou et. al. published a paper entitled “A Microfabricated device for Sizing and sorting DNA Molecules” in PNAS USA Vol. 96, pp. In Applied Physical Sciences, Biophysics, 11-13, Jan 1999, a microfabricated system is described to size and sort microscopic items based on a measurement fluorescent properties. The paper describes a method for measuring the length of DNA using fluorescent properties. This includes the amount of fluorescent dye intercalated within the DNA. In ?A Microfabricated Fluorescence-Activated Cells Sorter?, Nature Biotechnology, Vol. 17 November 1999, pages. A?T? A microfabricated structure was employed for cell sorting. The system used a detection window located upstream from the intersection of?T? The system used a detection window upstream of the?T? The fluid flow of a forward sorting system is changed based on a detected event. In reverse sorting, the fluid was initially set up to send all particles into a waste collection. However, upon detection of an event that could be collected, the fluid was reversed until the particle had been detected twice. Quake, et. al., PCT publication WO 99/61888 entitled “Microfabricated cell sorter” describes certain of these systems.
Others have tried to characterize biological system based on measuring different properties, such as electromagnetic radiation-related properties. Diverse efforts have been made to investigate dielectric properties of biological materials in the microwave range. See, e.g., Larson et al., U.S. Pat. No. No.
Despite the considerable effort made in the field, no comprehensive system that is effective, sensitive, and reliable has been achieved.
The methods and apparatus described here are based on the general use of light to extract information or apply forces from particles. Particles of any shape or size with a dielectric constant can be used. Optophoresis is the use of light to achieve these benefits. Optophoretic constants or signatures are used to identify a particle (such as a cell) and can be used for selection, sorting or characterization. The biological particles can be cells, organelles or proteins. They may also include any component at the atomic scale. These techniques are also applicable to non-biological matter such as metals, polymers, insulators and semiconductors.
In the biological world, the cell is the real point of integration for genomic information.” It is crucial to diagnose and treat disease by accessing and deciphering genomic information. The complexity of the information is too great for existing technologies to handle. “By unlocking the fundamental characteristics of the cell, the methods described herein creates new parameters for cell-based analysis, cellular characterization and cell assays.
This technology is a practical way to probe the inner workings a particle such as a cell. It’s best to do it without dyes, labels, or any other markers. The “Optophoretic Constant” The?Optophoretic Constant? These techniques enable the efficient and simple gathering of vast amounts of information. From screening new drugs to studying novel genes to developing new diagnostic products and monitoring cancer patients. This technology allows the simultaneous isolation and analysis of specific cells using this unique optophoretic characteristic. This technology can simultaneously analyze and isolate specific particles such as. Cells can be differentiated based on atomic differences. The technology can be used alone or combined with modern molecular techniques to identify the parameters of the complex mechanisms that govern the overall activity of the cell.
