Label-free detection of small and large molecule interactions, and activities in biological systems
First Claim
1. A system for quantitative detection and analysis of the binding of molecules with molecular receptors on the surfaces of a biological object, the system comprising:
- a microscope positioned to image the biological object and transmit a time sequence of images of the biological object;
a camera located to receive the transmitted time sequence of images of the biological object from the microscope and transmit data representing the time sequence of images;
a plurality of molecules introduced to bind with the molecular receptors;
a processor, coupled to receive the data representing the time sequence of images from the camera;
the processor including an image processing algorithm that quantitatively determines mechanical deformation of the biological object from the time sequence of images, wherein the mechanical deformation is due to an interaction of the biological object with the plurality of molecules, wherein the image processing algorithm determines mechanical deformation by using an edge detector to determine an edge of the biological object and a time sequence tracker for tracking a plurality of movements of the edge by measuring positions of the edge at different times, andwherein the image processing algorithm also determines molecular binding kinetic values of the biological object as proportional to the mechanical deformation of the biological object.
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Abstract
A system for quantitative detection and analysis of the interactions of molecules with molecular receptors on the surfaces of biological cells based on detecting a mechanical deformation in the membrane of a cell associated with the molecular interactions, which works for both large and small molecules. The mechanical deformation can be detected with high precision in real time from an optical image of the cell with a differential detection method. The system can be also used to detect the electrical activities, such as ion channel opening and closing, as well as action potential propagation in neuronal cells.
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Citations
23 Claims
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1. A system for quantitative detection and analysis of the binding of molecules with molecular receptors on the surfaces of a biological object, the system comprising:
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a microscope positioned to image the biological object and transmit a time sequence of images of the biological object; a camera located to receive the transmitted time sequence of images of the biological object from the microscope and transmit data representing the time sequence of images; a plurality of molecules introduced to bind with the molecular receptors; a processor, coupled to receive the data representing the time sequence of images from the camera; the processor including an image processing algorithm that quantitatively determines mechanical deformation of the biological object from the time sequence of images, wherein the mechanical deformation is due to an interaction of the biological object with the plurality of molecules, wherein the image processing algorithm determines mechanical deformation by using an edge detector to determine an edge of the biological object and a time sequence tracker for tracking a plurality of movements of the edge by measuring positions of the edge at different times, and wherein the image processing algorithm also determines molecular binding kinetic values of the biological object as proportional to the mechanical deformation of the biological object. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for quantitative detection and analysis of the binding of molecules with molecular receptors on the surfaces of a biological object, the method comprising:
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operating a microscope and a sensor to capture a time sequence of images of a biological object; introducing a plurality of molecules to interact with the biological object; operating a processor to quantitatively determine the mechanical deformation of the biological object from the time sequence of images due to interaction of the biological object with the plurality of molecules, wherein determining the mechanical deformation comprises operating an edge detector for determining an edge of the biological object and tracking a plurality of movements of the edge by measuring positions of the edge at different times; and operating the processor to determine molecular binding kinetic values of the biological object as proportional to the mechanical deformation of the biological object and to determine binding kinetic constants, including ka, kd and KD from the time sequence of images. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19)
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20. A system for quantitative detection and analysis of binding of neuron stimulator molecules with molecular receptors on the surfaces of a neuron, the system comprising:
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a microscope positioned to image the neuron and transmit a time sequence of images of the biological object; an image sensor located to receive transmitted images of the neuron from the microscope and transmit data representing the time sequence of images; neuron stimulator molecules introduced to bind with the molecular receptors; a processor coupled to receive the data representing the time sequence of images from the image sensor, the processor including an image processing algorithm that quantitatively determines mechanical deformation of the neuron from the time sequence of images, wherein the mechanical deformation is due to interaction of the neuron with the plurality of neuron stimulator molecules, wherein the image processing algorithm determines mechanical deformation by using an edge detector to determine an edge of the biological object and a time sequence tracker for tracking a plurality of movements of the edge by measuring positions of the edge at different times, and wherein the image processing algorithm also determines molecular binding kinetic values of the neuron as proportional to the mechanical deformation of the neuron. - View Dependent Claims (21)
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22. A method for quantitative detection and analysis of the binding of neuron stimulator molecules with molecular receptors on the surfaces of a neuron, the method comprising:
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operating a microscope and a sensor to capture a time sequence of images of a biological object; introducing a plurality of neuron stimulator molecules to interact with the neuron; operating a processor to quantitatively determine mechanical deformation of the neuron from the time sequence of images due to interaction of the neuron with the plurality of neuron stimulator molecules, wherein determining the mechanical deformation comprises operating an edge detector for determining an edge of the biological object and tracking a plurality of movements of the edge by measuring positions of the edge at different times; and operating the processor to determine molecular binding kinetic values of the neuron as proportional to the mechanical deformation of the neuron. - View Dependent Claims (23)
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Specification