Imaging electrophoresis system
Apparatus and method employing gel electrophoresis and optical imaging techniques to measure the amount of biomaterial that attaches to specified locations on a detector slide and to determine the molecular weight of the molecules at those locations.
1. Apparatus comprising a light source for emitting a beam of polarized light, said apparatus also including a light reflection surface arranged such that said light beam is reflected from the light reflection surface at an angle to provide an evanescent field above the light reflection surface, said light reflection surface having an array of probes placed thereon in said evanescent field, said apparatus also including a detector positioned to detect polarization changes in said reflected light beam caused by localized binding events between probes of said array and analyte molecules of a sample said apparatus including a gel layer above said probes; and
a first means for generating an electric field in said gel layer extending laterally with respect to said reflection surface for moving by electrophoresis functioning molecules of a sample introduced to said gel layer laterally with respect to the array of probes.
2. Apparatus as in claim 1 said array of probes having a plurality of probe patterns in which each probe pattern is functional for binding detection of a different analyte.
3. Apparatus as in claim 1 also including a second means for generating a second electric field perpendicular to said first electric field.
4. Apparatus as in claim 1 wherein said array of probes is located in a first portion of said light reflection surface spaced laterally apart from a sample supply area and said electric field is operative in a second portion of said light reflection surface between said sample supply area and said first portion to separate by molecular weight in said gel layer the molecules of a sample introduced at said sample supply area.
5. A system for enhancing chemical detection and analysis comprising:
a transparent substrate having a top surface:
a gel layer over the top surface;
a source of an electric field operative by electrophoresis functioning to move molecules within the gel layer along one or more selected paths laterally with respect to the top surface;
one or more arrays of probe patterns on the top surface;
wherein the one or more selected paths are determined based on selected proximity to the one or more arrays of probe patterns;
whereby molecules being moved by electrophoresis are caused to traverse a path proximate the one or more arrays of probe patterns in order to enhance the occurrence of binding events; and
an apparatus to effect total internal reflection imaging ellipsometry to create an evanescent field above the top surface such that the probe arrays are in the evanescent field said apparatus using a source of polarized light to enable detection of polarization state information caused by binding events to the probe array.
6. The system of claim 5 further comprising:
a plurality of probe patterns on the substrate each probe pattern being functional for binding detection of a single specific molecule;
the probe patterns being arranged in an order relative to the electrophoresis functioning path of movement of molecules such that the molecules will be arranged in the same order.
7. The system of claim 5 further comprising:
a light source for emitting a beam of polarized light, arranged such that said light beam is reflected from the top surface in a manner to provide an evanescent field above the top surface such that the probes are in the evanescent field; and
a polarization sensitive imaging system arranged to receive and process the reflected light whereby binding events can be imaged.
8. The system of claim 7 further comprising;
a plurality of probe patterns on the substrate each probe pattern being functional for binding detection of a single specific molecule;
the probe patterns being arranged in an order relative to the electrophoreses functioning path of movement of molecules such that the molecules will be arranged in the same order;
wherein binding events will be organized according to the order of arrangement of probes.
9. The system of claim 5 further wherein said electric field defines a first electric field operative to move molecules through the gel layer in a first lateral direction relative to the array patterns and further comprising a second electric field at a right angle to the first electric field operative to move molecules in a second lateral direction relative to the array patterns which is at a right angle to the first lateral direction.
10. The system of claim 5 further wherein said electric field defines a first electric field operative to move molecules through the gel layer in a first lateral direction relative to the array patterns and further comprising an additional electric field operative to cause said molecules to traverse a path in a vertical direction relative to the array patterns.
11. A cassette including a slide having a top and a bottom surface, an array of probes on at least a portion of said top surface and a gel layer on said probes, said cassette also including means for generating an electric field in said gel layer for moving analyte molecules in said gel layer laterally with respect to said probes, wherein said slide is transparent and operative to transmit light from said bottom to said top surface in a manner to generate an evanescent field above said top surface.
12. A system for enhancing chemical detection and analysis comprising:
a transparent substrate having a top surface:
a gel layer on the top surface;
a source of an electric field operative by electrophoresis functioning to move molecules within the gel layer along one or more selected paths laterally with respect to the top surface;
one or more arrays of probe patterns on the top surface;
wherein the one or more selected paths are determined based on selected proximity to the one or more arrays of probe patterns;
whereby molecules being moved by electrophoresis are caused to traverse a path proximate the one or more arrays of probe patterns in order to enhance the occurrence of binding events; and
an imaging apparatus comprising;
a light source emitting a polarized beam of light said light source being directed at the transparent substrate to be totally internally reflected at the top surface and to provide an evanescent field such that the one or more arrays of probes are in the evanescent field such that the one or more array of probes in the evanescent field causes spatially distributed polarization changes in the cross-section of the light beam; and
a detector positioned to detect the spatially distributed polarization changes caused by the probes.