Automated sample analysis

• US 7,595,197 B2
• Filed: 05/10/2004
• Issued: 09/29/2009
• Est. Priority Date: 05/09/2003
• Status: Active Grant

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Inventors: Brasseur, Michael
• Primary Examiner(s): Warden; Jill
• Assistant Examiner(s): TURK, NEIL N
• Application Number:
  US10/842,073
• Publication Number:
  US 20050003554A1
• Time in Patent Office: 1,968 Days
• Field of Search: None
• US Class Current: 436/172
• CPC Class Codes: B01F 25/31 in conduits or tubes through which the main component flows
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/148 Specific details about calibrations
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• CPC Class Codes: 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 B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 9/527 for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: G01N 2021/6443 Fluorimetric titration
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 2035/00148 Test cards, e.g. Biomerieux or McDonnel multiwell test cards
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 21/276 with alternation of sample and standard in optical path
• CPC Class Codes: G01N 21/643 : non-biological material
• CPC Class Codes: G01N 21/645 : Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/79 : Photometric titration
• CPC Class Codes: G01N 27/44721 : by optical means
• CPC Class Codes: G01N 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 35/00029 : provided with flat sample substrates, e.g. slides G01N35/028 takes precedence
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

Samples of materials used in industrial processes are analyzed to determine the concentration of certain materials of interest. The quantitative analysis of samples for these materials is provided without the need for manual methods such as titration. Indicators such as fluorescent dyes for which the intensity of fluorescence is indicative of the concentration of a material of interest are used. The dyes are made to fluoresce by means of a light source, and a photomultiplier or other detector capable of measuring light intensity detects the resulting fluorescence. The intensity of fluorescence in the sample is compared to the intensities of fluorescence produced by samples with known concentrations of the material of interest to determine the concentration of the material of interest of the sample.

Claims

1. A method of determining unknown concentrations of an inorganic ion of interest in a recovered sample comprising the steps of:

providing a microfluidic device comprising a sample well having the recovered sample disposed therein, an indicator well having a fluorescent indicator disposed therein, a control well having a control disposed therein, the control including a known quantity of the inorganic ion of interest, a waste well, a detection region, and a light source directed at the detection region;

wherein the sample well, the indicator well, and the waste well form a first fluidic circuit, the sample well fluidly coupled to the waste well via a first channel, and the indicator well fluidly connected to the first channel via a second channel;

and wherein the indicator well, the control well, and the waste well form a second fluidic circuit, the indicator well fluidly coupled to the waste well via a third channel, and the control well fluidly coupled to the third channel via fourth, fifth, and sixth channels, each of the fourth, fifth, and sixth channels having a different flow resistance;

mixing, in the third channel, the control sample and the fluorescent indicator, wherein flowing the control sample into the third channel via the fourth, fifth, and sixth channels provides a plurality of relative known ratios of the control sample in the indicator;

obtaining a signal representative of the concentration of the inorganic ion of interest in each of the ratios, wherein the signal comprises an intensity of fluorescence;

generating a model representative of the signal and the relative ratio of the control sample in the indicator;

mixing, in the first channel, the recovered sample and the fluorescent indicator, and obtaining a signal representative of the concentration of the inorganic ion of interest therein, wherein the signal comprises an intensity of fluorescence; and

comparing the signal representative of the inorganic ion of interest for the recovered sample within the model, thereby providing a determination of the concentration of the inorganic ion of interest present in the recovered sample.

2. The method of claim 1, wherein the signal indicative of the inorganic ion of interest concentration is an intensity of fluorescence, and the model is a graphical representation of the fluorescence intensity vs. concentration of the inorganic ion of interest.

3. The method of claim 1 wherein the microfluidic device further comprises a buffer well having a buffer disposed therein, the buffer well, the sample well, the indicator well, and the waste well forming a third fluidic circuit, the sample well further fluidly coupled to the waste well via a seventh channel, the buffer well fluidly coupled to the seventh channel via an eighth channel, and the indicator well further fluidly connected to the seventh channel via a ninth channel, the method further including the step of mixing, in the seventh channel, a combination of recovered sample, buffer, and fluorescent indicator and passing the combination through the detection region for analysis.

4. The method of claim 1 wherein the microfluidic device further comprises a second sample well having a second recovered sample disposed therein, the second sample well, the indicator well, and the waste well forming a fourth fluidic circuit, the second sample well fluidly coupled to the waste well via a tenth channel, and the indicator well further fluidly connected to the tenth channel via an eleventh channel, the method further including the steps of:

flowing the second recovered sample from the second sample well;

mixing, in the tenth channel, the second recovered sample with the buffer and the fluorescent indicator; and

obtaining a signal indicative of the concentration of an inorganic ion of interest in this mixture, wherein the signal comprises an intensity of fluorescence.

5. The method of claim 1, wherein pressure is applied to at least one well of the device to cause each of the mixing steps.

6. The method of claim 5, wherein the pressure is a vacuum.

7. The method of claim 5, wherein the pressure is above atmospheric.