AUTOMATED SYNTHESIS OR SEQUENCING APPARATUS AND METHOD FOR MAKING AND USING SAME
First Claim
Patent Images
1. An apparatus comprising:
- a continuous substrate including zones formed therein and/or thereon, each zone including one binding agent or a sparsely distributed plurality of binding agents;
a component introduction stations adapted to introduce one or a plurality of components onto and/or into one or a plurality of zones of the substrate, where one or more of the components are adapted to interact or bond to the binding agents to form one or a sparsely distributed immobilized active sites, where the binding agents, one or more of the components and/or the substrate include at least one detectable agents, each agent having a detectable property and where each agent or property are the same or different;
a detection station adapted to detect reaction and/or interaction events occurring at the distinct and detectable active sites within a viewing field associated with each zone over a desired period of time, where the detection station includes a detector adapted to produce output signals corresponding to the detected events;
an analyzer adapted to receive the signals from the detection station and to convert the signals into output data characterizing the detected events occurring within each field over the period of time;
a means for moving the substrate to bring a new zone or plurality of zones to the introduction stations and the detections stations until a desired length of substrate has been processed.
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Abstract
An apparatus and method based on the apparatus is disclosed for automated single molecule or molecular assemblage detection via light irradiation and detection of transient FRET between a donor or acceptor bound to an immobilized single molecule or molecular assemblage and a corresponding acceptor or donor associated with, covalently bonded to, a reagent, where the donor or acceptor associated with the reagent is transiently in FRET proximity to the acceptor or donor associated with the immobilized molecule or molecular assemblage.
105 Citations
26 Claims
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1. An apparatus comprising:
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a continuous substrate including zones formed therein and/or thereon, each zone including one binding agent or a sparsely distributed plurality of binding agents; a component introduction stations adapted to introduce one or a plurality of components onto and/or into one or a plurality of zones of the substrate, where one or more of the components are adapted to interact or bond to the binding agents to form one or a sparsely distributed immobilized active sites, where the binding agents, one or more of the components and/or the substrate include at least one detectable agents, each agent having a detectable property and where each agent or property are the same or different; a detection station adapted to detect reaction and/or interaction events occurring at the distinct and detectable active sites within a viewing field associated with each zone over a desired period of time, where the detection station includes a detector adapted to produce output signals corresponding to the detected events; an analyzer adapted to receive the signals from the detection station and to convert the signals into output data characterizing the detected events occurring within each field over the period of time; a means for moving the substrate to bring a new zone or plurality of zones to the introduction stations and the detections stations until a desired length of substrate has been processed. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24. An apparatus comprising:
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a continuous substrate including zones formed therein and/or thereon, each zone including one binding agent or a sparsely distributed plurality of binding agents; a component introduction stations adapted to introduce one or a plurality of components onto and/or into one or a plurality of zones of the substrate, where one or more of the components are adapted to interact or bond to the binding agents to form one or a sparsely distributed immobilized active sites, where the binding agents, one or more of the components and/or the substrate include at least one detectable agents, each agent having a detectable property and where each agent or property are the same or different; a mapping station to locate or map distinct and detectable pre-active sites inside the zones relative to a detection grid superimposed on the zones; an initiation station adapted to introduce one or a plurality of initiation reagents onto and/or into the zones; a detection station adapted to detect reaction and/or interaction events occurring at the distinct and detectable active sites within a viewing field associated with each zone over a desired period of time, where the detection station includes a detector adapted to produce output signals corresponding to the detected events; and an analyzer adapted to receive the signals from the detection station and to convert the signals into output data characterizing the detected events occurring within each field over the period of time; a means for moving the substrate to bring a new zone or plurality of zones to the introduction stations and the detections stations until a desired length of substrate has been processed.
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25. A method for analyzing one reaction site, a small ensemble of reaction sites, a medium ensemble of reaction sites and a large ensemble of reaction sites comprising the step of:
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providing a continuous substrate including a zone, where the zone includes one or a plurality of sparsely distributed binding sites; passing the continuous substrate through one or a plurality of component introduction stations adapted to introduction the components required to immobilize and produce active sites in the zones, each site including at least one detectable agent having a detectable property, where the agents and the properties are the same or different; passing the continuous substrate including active sites through a detection system, where reactions and/or interactions occurring at the sites within a viewing field are detected in a detector of the detection system to produce detected event signals, and analyzing the detected event signals to convert the signals into data about the detected events.
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26. A method for analyzing one reaction site, a small ensemble of reaction sites, a medium ensemble of reaction sites and a large ensemble of reaction sites comprising the step of:
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providing a continuous substrate including a zone, where the zone includes one or a plurality of sparsely distributed binding sites; passing the continuous substrate through one or a plurality of component introduction stations adapted to introduce components required to immobilize and produce pre-active sites in the zones, each site including at least one detectable agent having a detectable property, where the agents and the properties are the same or different; passing the continuous substrate including the pre-active sites through a mapping station, where the pre-active sites are mapped relative to a grid associated with a viewing field of the mapping detector, passing the continuous substrate including the pre-active sites through an initiation station, where one or a plurality of initiators are introduced into or onto the zones to convert some or all of the pre-active sites into active sites within the zones; passing the continuous substrate including the active sites through a detection system, where reactions and/or interactions occurring at the sites within a viewing field are detected in a detector of the detection system to produce detected event signals, and analyzing the mapped and detected event signals to convert the signals into data about the detected events.
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Specification