Detecting target molecules in a sample
First Claim
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1. A method of detecting the presence of a luminescently labeled target molecule in a sample, the method comprising acts of:
- providing a substrate having immobilized thereon probe molecules that specifically bind to the luminescently labeled target molecule to result in binding complexes bound to the substrate;
flowing the sample through the substrate by introducing a stream of the sample to one side of the substrate, the stream passing through and out of an opposing side of the substrate forming the binding complexes that are bound throughout the substrate while leaving unbound luminescently labeled target molecules;
pressing a wash fluid through the substrate by introducing a stream of the wash fluid under pressure at the one side of the substrate, the stream of the wash fluid passing through the substrate and out of the opposing side of the substrate, the wash fluid having a refractive index within a range of plus and minus 6% of a refractive index of the substrate;
while the substrate is in the wash fluidremoving the unbound luminescently labeled target molecules from the substrate,irradiating the substrate throughout with a beam of radiation, anddetecting an emitted luminescence from the luminescently labeled target molecules throughout the substrate which is indicative of absence or presence of the binding complexes on and within the substrate; and
determining from the luminescence emitted throughout the substrate whether the luminescently labeled target molecule is present in the sample.
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Abstract
The invention relates to detection the presence of a target molecule in a sample, wherein the sample is contacted with a substrate, the substrate subsequently being washed in a wash step. In particular, the invention relates to a method of detecting the presence of a target molecule in a sample, the method comprising: (a) contacting the sample (37) with a substrate having immobilized thereon probe molecules that specifically binds to the target molecule; (b) washing the substrate (38) in a wash step by a wash fluid in order to remove or dilute unbound target molecules; (c) detect the presence of resultant binding complexes (39) on the substrate to determine whether the target molecule is present in the sample. The wash fluid being substantially refractive index matched to the substrate.
12 Citations
18 Claims
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1. A method of detecting the presence of a luminescently labeled target molecule in a sample, the method comprising acts of:
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providing a substrate having immobilized thereon probe molecules that specifically bind to the luminescently labeled target molecule to result in binding complexes bound to the substrate; flowing the sample through the substrate by introducing a stream of the sample to one side of the substrate, the stream passing through and out of an opposing side of the substrate forming the binding complexes that are bound throughout the substrate while leaving unbound luminescently labeled target molecules; pressing a wash fluid through the substrate by introducing a stream of the wash fluid under pressure at the one side of the substrate, the stream of the wash fluid passing through the substrate and out of the opposing side of the substrate, the wash fluid having a refractive index within a range of plus and minus 6% of a refractive index of the substrate; while the substrate is in the wash fluid removing the unbound luminescently labeled target molecules from the substrate, irradiating the substrate throughout with a beam of radiation, and detecting an emitted luminescence from the luminescently labeled target molecules throughout the substrate which is indicative of absence or presence of the binding complexes on and within the substrate; and determining from the luminescence emitted throughout the substrate whether the luminescently labeled target molecule is present in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method of detecting the presence of a luminescent target molecule in a sample, the method comprising:
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contacting the sample with a substrate having immobilized thereon probe molecules that specifically binds to the luminescent target molecule; washing the substrate by a wash fluid in order to remove or dilute unbound luminescent target molecules; detecting the presence of resultant binding complexes on the substrate to determine whether the luminescent target molecule is present in the sample; wherein the substrate is irradiated with a beam of radiation and subsequently the resulting luminescence beam from the substrate is detected and the presence of the luminescent target molecules is detected by detecting luminescence of the target molecules Wherein the irradiation with a beam of radiation and the detecting of the resulting luminescent beam from the substrate takes place in the presence of the wash fluid where the refractive index of the wash fluid is within the range of the refractive index of the substrate plus 10% to the refractive index of the substrate minus 10%. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18)
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Current AssigneeKoninklijke Philips N.V.
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Original AssigneeKoninklijke Philips N.V.
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InventorsVan Herpen, Maarten Marinus Johannes Wilhelm, Klunder, Derk Jan Wilfred, Stapert, Hendrik Roelof
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Primary Examiner(s)Boesen, Christian C
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Application NumberUS12/446,461Publication NumberTime in Patent Office4,015 DaysField of SearchNoneUS Class CurrentCPC Class CodesC40B 30/10 by measuring physical prope...C40B 60/04 Integrated apparatus specia...G01N 21/6428 Measuring fluorescence of f...G01N 33/5306 Improving reaction conditio...
