# US 11,441,173 B2 · Optical instruments and systems for forensic DNA quantitation

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- Number: US 11,441,173 B2
- Document ID: US11441173B2
- Status: Granted
- Kind: B2
- Application number: 16/729,224
- Filed: 2019-12-27
- Published: 2020-08-06
- Granted: 2022-09-13
- Earliest claimed: 2009-06-15
- Assignees: ANDE Corporation
- Applicants: ANDE Corporation
- Inventors: Eugene Tan; Richard F. Selden; Rosemary S. Turingan
- Web page: https://patentgrep.com/patent/US11441173B2

## CPC

- C12Q 1/6825: Nucleic acid detection involving sensors
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- C12N 15/1017: {by filtration, e.g. using filters, frits, membranes}
- C12Q 1/6816: characterised by the detection means (C12Q1/6804 takes precedence)
- C12Q 1/6818: involving interaction of two or more labels, e.g. resonant energy transfer
- B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip}
- B01L 2300/0627: Sensor or part of a sensor is integrated
- B01L 2300/0654: Lenses; Optical fibres
- B01L 2300/0819: Microarrays; Biochips
- B01L 2300/0877: Flow chambers
- B01L 2300/168: Specific optical properties, e.g. reflective coatings
- G01N 15/1484: {microstructural devices}
- G01N 2201/06113: Coherent sources; lasers
- G02B 27/10: Beam splitting or combining systems (polarising systems G02B27/28; mixing and splitting light signals using optical waveguides G02B6/28; {beam shaping, e.g. changing the cross-sectional area, by dividing or superposing multiple light beams G02B27/0905})
- Y10T 436/143333: Saccharide [e.g., DNA, etc.]
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

Described herein are instruments for excitation and detection of fluorophores in a plurality of functional regions in a biochip, using an excitation source and a steering element that directs a beam from the excitation source to a plurality of functional regions in the biochip, wherein the excitation source excites the fluorophores in the plurality of functional regions generating a signal that is detected such that said signal from at least one of the plurality of functional regions allows for nucleic acid quantification. Also described are systems for quantification and separation and detection using optical devices adapted for preliminary, simultaneous or sequential quantitation of nucleic acid in separate detection positions, and for the excitation and detection of multiple samples to steer both the excitation and detection beam paths to separately image each lane of a biochip.

## Claims

### Claim 1

A system comprising: (a) a biochip comprising a plurality of microfluidic systems, wherein each microfluidic system comprises one or a plurality of first reaction chambers, each adapted for microfluidic quantitation, wherein each of said plurality of said first reaction chambers further comprises a first detection position, and a separation chamber, wherein the separation chamber comprises a second detection position;
(b) a data acquisition and storage system;
(c) a quantitation system, comprising;
(i) one or a plurality of first reaction chambers, each adapted configured for a quantitation assay or method that determines the amount of a measurable characteristic which may be correlated to the amount of said one or a plurality of target analytes in the quantitation chamber;
(ii) one or more light sources positioned to illuminate the first detection positions on the biochip;
(iii) one or a plurality of first optical elements positioned for collecting and directing light emanating from said first detection positions; and
(iv) a light detector positioned to accept light directed from the first optical elements, wherein the light detector comprises a wavelength dispersive element configured to separate the light from the first optical elements according to light wavelength and positioned to provide a portion of the separated light to at least one detection elements, wherein each of the detection elements are in communication with said data acquisition and storage system and are configured for simultaneously collecting detection information from each of the detection elements; and wherein said light detector being is configured to detect fluorescence from at least one dye labeled to one or more biological molecules to be quantitated, each dye having a unique peak emission wavelength, where the measurable characteristic is correlated to the amount of said one or a plurality of target analytes in the quantitation chambers;
(d) a separation and detection system, comprising;
(i) a separation element configured for simultaneously separating one or a plurality of target analytes in the separation chamber;
(ii) one or more light sources positioned to illuminate the second detection positions on the biochip;
(iii) one or a plurality of first optical elements positioned for collecting and directing light emanating from said second detection positions; and
(iv) a light detector positioned to accept light directed from the first optical elements, wherein the light detector comprises a wavelength dispersive element configured to separate the light from the first optical elements according to light wavelength and positioned to provide a portion of the separated light to at least one detection element, wherein each of the detection elements are in communication with said data acquisition and storage system and are configured for simultaneously collecting detection information from each of the detection elements; and wherein said light detector being is configured to detect fluorescence from at least one dye labeled to one or more biological molecules, each dye having a unique peak emission wavelength, where the separation element is for simultaneously separating one or a plurality of target analytes in the separation chambers; and
(e) a translational mirror in communication with a first control element and configured for setting a first excitation and detection beam path for said quantitation system or a second excitation and detection beam path for said separation and detection system.

### Claim 2 (depends on 1)

The system of claim 1 wherein the translational mirror is used for preliminary, simultaneous or sequential quantitation of nucleic acid in each of the detection positions, and for the excitation and detection of multiple samples to steer both the excitation and detection beam paths in order to image each of the lanes of the biochip.

### Claim 3 (depends on 1)

The system of claim 1 wherein the quantitation system uses an excitation beam that is directed to a lens, a first scanning mirror and a lens and objective assembly.

### Claim 4 (depends on 1)

The system of claim 1 wherein in element (c)(1) said one or a plurality of first reaction chambers, is each adapted for a quantitation assay or method that determines the amount of a measurable characteristic which may be correlated to the amount of said one or a plurality of target analytes in the quantitation chamber without prior amplification.

### Claim 5 (depends on 1)

The system of claim 1 wherein the separation element comprises a set of dichroic mirrors and a bandpass filters or a spectrograph or a prism.

### Claim 6 (depends on 1)

The system of claim 1 wherein the light detector comprises a PMT, a multi-element PMT, a multielement photodiode a CCD camera a back sided CCD camera or a multi-element PMT.

### Claim 7 (depends on 6)

The system of claim 6 wherein the light detector is a 32 element PMT detector array.

### Claim 8 (depends on 1)

The wavelength of claim 1 where dispersive elements comprise prisms, diffraction gratings, transmission gratings and spectrographs.