Massively parallel detection

• US 5,854,684 A
• Filed: 09/26/1996
• Issued: 12/29/1998
• Est. Priority Date: 09/26/1996
• Status: Expired due to Fees

• Current Assignee: Sarnoff Corporation (SRI International, Inc.)
• Original Assignee: Sarnoff Corporation (SRI International, Inc.)
• Inventors: Rosen, Arye, Zanzucchi, Peter John, Heaney, Paul, Ludington, David Norman, Cherukuri, Satyam Choudary, York, Pamela Kay, Stabile, Paul J.
• Primary Examiner(s): STUCKER, JEFFREY J
• Application Number:
  US08/721,432
• Time in Patent Office: 824 Days
• Field of Search: 422/82.05, 436/518, 356/246, 356/343, 356/369, 356/440, 356/73, 356/417, 369/112, 369/44.12, 369/44.14, 369/44.23, 369/44.24, 369/44.37, 250/559.01, 250/559.09, 250/206.1, 250/208.1, 349/70, 349/57
• US Class Current: 356/440
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00434 Liquid crystal masks
• CPC Class Codes: B01J 2219/00448 using microlens arrays
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00

Abstract

The invention provides apparatuses for detecting light from, for example, closely spaced detection sites. In one embodiment, the invention provides an apparatus for measuring the amount of light emitted from a first set of two or more detection sites on a planar substrate while spatially resolving the measurements from each first set detection site, the apparatus comprising: a source of a light beam directed towards the planar substrate at a first angle; one or more lenses for focusing light emitted or reflected from each of the first set detection sites and having a second angle having an angle offset from the first angle, onto a unique area of an array detector; and the array detector comprising a plurality of light responsive pixels, wherein for each first detection site there is at least one light responsive pixel that receives light emitted or reflected from that detection site and substantially no cross-talk from another detection site, and wherein substantially none of the light from the light source intersects with the array detector.

Claims

1. An apparatus for measuring the amount of light emitted or reflected from or transmitted through a first set of two or more detection sites on a planar substrate while spatially resolving the measurements from each first set detection site, the apparatus comprising:

a source of a light beam directed towards the planar substrate at a first angle;

one or more lenses for focusing light emitted or reflected from or transmitted through each of the first set detection sites and having a second angle having an angle offset from the first angle, onto a unique area of an array detector; and

the array detector comprising a plurality of light responsive pixels, wherein for each of the first set detection sites there is at least one light responsive pixel that receives light emitted or reflected from or transmitted through that detection site and substantially no cross-talk from another detection site, and wherein substantially none of the light from the light source intersects with the array detector.

2. The apparatus of claim 1, wherein the detector array has sufficient light responsive pixels and is designed to work with a planar substrate having sufficient separation between the detection sites so that

(a) there are first light responsive pixels of the detector array that are aligned to receive light emitted or reflected from or transmitted through the first set detection sites and

(b) there are second light responsive pixels of the detector array that receive substantially no light because they are aligned with an area of blocking material,

such that for each first set detection site there is a grouping of one or more first light responsive pixels receiving light therefrom and this grouping is separated from the grouping for any other first set detection site by at least one second light responsive pixel.

3. The apparatus of claim 2, wherein the detector array has sufficient light responsive pixels aligned with each first set detection site so that the light emitted or reflected from or transmitted through the detection site can be spatially resolved to detect differences across the detection sites in the quantity of light emitted or reflected therefrom.

4. The apparatus of claim 1, wherein the array detector comprises a photon-based imaging device.

5. The apparatus of claim 4, comprising the array detector which is selected from the group consisting of a charge coupled device (CCD), an intensified CCD array, a focal plane array, a photodiode array or photodetector array.

6. The apparatus of claim 1, wherein the apparatus is designed for use with planar substrates having detection sites that are concave depressions for holding liquid, wherein the surfaces on the depressions have a coating of one or more layers of material, wherein the coating is designed to reflect the light emitted or reflected from or transmitted through the detection sites.

7. The apparatus of claim 6, wherein the light source is designed to position beneath the planar substrate and the coating is designed to transmit the source light.

8. The apparatus of claim 1, wherein the apparatus is designed to detect light emitted or reflected from or transmitted through at least about 100 detection sites of the first set and has at least one light responsive pixel aligned with each detection site of the first set.

9. The apparatus of claim 1, wherein the source of light comprises at least one light producing device per detection site.

10. The apparatus of claim 1, wherein the one or more focusing lenses comprise a separate lenslet overlaid on the light responsive pixels aligned with each first set detection site.

11. The apparatus of claim 1, wherein the apparatus further comprises

an electronic storage device comprising a plurality of storage registers coupled to the array detector for storing the data from the array detector;

a processor having access to the stored data; and

a motor for moving the substrate, light source or array detector under the control of the processor,

wherein the processor is programmed to use one or more initial illuminations of the substrate with the source light to generate data which the processor uses to operate the motor to correct the alignment of the light source, first set detection sites and array detector.

12. The apparatus of claim 1, wherein the offset between the first and second angles is from at least about 10°

to 180°

.

13. An assay system comprising the apparatus of claim 1 and a plate having a first edge and a second edge and having at least about 1,000 uniformly sized reaction cells formed in its upper surface, wherein the density of the reaction cells is at least about 10 cells per cm², wherein the apparatus is designed to detect light emitted or reflected from or transmitted through the uniformly sized reaction cells.

14. An apparatus for measuring the amount of light emitted from a first set of two or more detection sites on a planar substrate while spatially resolving the measurements from each first set detection site, the apparatus comprising:

a source of a light beam having a first wavelength directed towards the planar substrate at a first angle;

one or more lenses for focusing light emitted from each of the first set detection sites and having a second angle onto a unique area of an array detector;

a filter interposed between the detection sites and the array detector, which filter selectively absorbs light of the first wavelength and transmits light emitted from the detection sites having a wavelength differing from the first wavelength; and

the array detector comprising a plurality of light responsive pixels, wherein for each first detection site there is at least one light responsive pixel that receives light emitted from that detection site and substantially no cross-talk from another detection site, wherein substantially none of the light from the light source intersects with the array detector.

15. The apparatus of claim 14, further comprising:

a first polarizing filter for polarizing the source light beam to a first polarity; and

a second polarizing filter for polarizing the light emitted from the detection sites to a second polarity, which is offset from the first polarity.

16. A method for measuring and spatially resolving the amount of light transmitted through a first set of two or more detection chambers or the amount of light emitted from the first set detection sites as a result of their illumination, wherein each chamber is separated by a blocking material that is opaque to the light and the density of first set detection sites is at least about 10 per cm², the method comprising

(1) providing an apparatus comprising:

a source of light directed towards the planar substrate at a transmission angle;

one or more lenses for focusing light from each first set detection chamber onto a unique area of an array detector; and

the array detector comprising a plurality of light responsive pixels, wherein for each first set detection chamber there is at least one light responsive pixel that receives light transmitted through that detection chamber and substantially no cross-talk from another detection chamber, and

(2) measuring light transmitted through the detection sites using the apparatus.

17. The method of claim 16, comprising operating the source of light to direct a pulse of light towards the first set detection sites and beginning to collect a light response in the array detector after the end of the light pulse while the source of light is not producing light.

18. The method of claim 16, wherein the apparatus is designed for use with planar substrates having detection sites that are concave depressions for holding liquid, wherein the surfaces on the depressions have a coating of one or more layers of material, wherein the coating is designed either to reflect the light emitted or transmitted from the detection sites or transmitted from the source of light.

19. An apparatus for measuring and spatially resolving the amount of light emitted from a first set of two or more detection sites arranged on a planar substrate with a density of at least about 10 detection sites per cm², the apparatus comprising:

one or more lenses for focusing light emitted from the first set detection sites, which light has a detection angle, onto an array detector; and

the array detector comprising a plurality of light responsive pixels, wherein for each detection site there is at least one light responsive pixel that receives light emitted from that detection site and substantially no cross-talk from another detection site.

20. The apparatus of claim 19, further comprising the planar substrate, wherein either the planar substrate incorporates a scintillate adjacent to each detection site or the detection sites comprise chambers suitable for holding a fluid containing a scintillate.