Device for optical and electrochemical measurements in microliter size samples

• US 6,043,878 A
• Filed: 07/09/1999
• Issued: 03/28/2000
• Est. Priority Date: 09/24/1996
• Status: Expired due to Term

• Current Assignee: Case Western Reserve University
• Original Assignee: Case Western Reserve University
• Inventors: Gratzl, Miklos, Cserey, Andrea, Hui, Kwong Yue
• Primary Examiner(s): Font, Frank G.
• Assistant Examiner(s): STAFIRA, MICHAEL PATRICK
• Application Number:
  US09/147,986
• Time in Patent Office: 263 Days
• Field of Search: 352/246, 352/244, 352/36, 352/38, 250/341, 250/343, 250/341.1
• US Class Current: 356/246
• CPC Class Codes: B01F 33/3021 the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
• CPC Class Codes: B01F 33/30351 using hydrophilic/hydrophobic surfaces
• CPC Class Codes: B01F 33/3039 with mixing achieved by diffusion between layers
• CPC Class Codes: B01F 33/407 by blowing gas on the material from above
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2400/0472 Diffusion
• 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 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 2035/00287 movable lid/cover for sample or reaction tubes
• CPC Class Codes: G01N 2035/1034 Transferring microquantities of liquid
• CPC Class Codes: G01N 21/79 Photometric titration

Abstract

A small liquid sample (24) to be tested is contained within an annulus (18) on the upper surface (14) of a substrate (12). In one embodiment, a reagent diffuses into the sample through a membrane (28) in a junction hole (26), the junction hole connecting upper (14) and lower (16) surfaces of the substrate. Optical measuring equipment (70, 72) detects a measurable change in an optical property of the sample. In an alternate embodiment, a flow of gas (30), directed at the small liquid sample (24), causes the sample to flow in a controlled manner over the surface of an electrode (20), disposed on the substrate surface (14). In another alternate embodiment, a special averaging electrode (20) is disposed in a non-homogeneous sample (24). Highly reproducible and accurate hydrodynamic electrochemical studies and analyses of microliter size samples are thus achievable, without the need for moving mechanical parts.

Claims

1. An apparatus for performing measurements on a small liquid sample including:

a substrate, including an upper surface for supporting the sample to be investigated, and a lower surface;

a container, sealed to the upper surface of the substrate for containing the sample on the substrate;

the container including an annulus that confines the sample on an area of the substrate upper surface bounded by the annulus;

a junction hole, passing through the substrate and connecting the upper surface of the substrate with a lower surface of the substrate,

a membrane covering the junction hole, the membrane permitting diffusive movement of a reagent through the junction hole and into the sample;

a source of reagent, disposed adjacent to the lower surface of the substrate for supplying reagent to the membrane; and

,

a detector for detecting a measurable change in a property of the sample.

2. The apparatus of claim 1, wherein:

the detector includes optical measuring equipment, the optical measuring equipment including a light source for directing light toward the sample and a receiver for receiving light from the sample.

3. The apparatus of claims 2, wherein:

the source of reagent includes a window, the window located such that light from the light source passes through the window and through both the reagent and the substrate to the sample.

4. The apparatus of claim 2, wherein:

the substrate includes a reflective coating, the coating reflecting light from the light source into the sample.

5. The apparatus of claim 1 further including:

a mixing system for mixing the sample;

the mixing system including a source of gas for directing a controlled flow of gas at the sample, the flow of gas rotating and mixing the sample as desired.

6. The apparatus of claim 1 wherein:

the membrane includes a voltage sensitive gel for controlling the rate of reagent diffusion through the membrane by applying a voltage across the gel.

11. A method for performing measurements on a small liquid sample, the method comprising:

disposing a sample to be tested in the apparatus of claim 1 by depositing the sample on the upper surface of the substrate, in the area bounded by the annulus;

adding a reagent to the droplet by diffusion of the reagent through the membrane;

making measurements on the sample, the measurements corresponding to a property of the sample.

16. The method of claim 11 wherein the property of the sample is the chemical composition of the sample.

7. An apparatus for performing hydrodynamic electrochemical studies and analyses on a small liquid sample including:

a substrate including, an upper surface for supporting the sample to be investigated;

an electrode, electrically connected to the sample;

a container, sealed to the upper surface of the substrate for containing the sample on the substrate,

the container including an annulus which confines the sample on an area of the substrate upper surface bounded by the annulus;

a source of gas, for directing a flow of gas toward the sample, thereby causing controlled liquid flow over the electrode.

8. The apparatus of claim 7, wherein:

the electrode includes a gold ring working electrode, centered on the area of the upper surface of the substrate bounded by the annulus.

9. The apparatus of claim 7 including:

a junction hole, passing through the substrate and connecting the upper surface of the substrate with a lower surface of the substrate;

a gel disposed in the junction hole;

a second electrode being disposed in an electrolyte, the electrolyte contacting the gel at the lower surface of the substrate such that the electrode is in electrical contact with the sample.

10. The apparatus of claim 7, including:

a reagent in solution, the solution contacting the gel at the lower surface of the substrate, such that the reagent diffuses through the gel and into the sample.

12. A method for performing hydrodynamic electrochemical studies and analyses on a small liquid sample, the method comprising:

disposing a sample to be tested in the apparatus of claim 7 by depositing the sample on the upper surface of the substrate, in the area bounded by the annulus;

directing a flow of gas onto the sample, thereby causing controlled liquid flow over the electrode;

conducting electrochemical studies on the sample.

13. An apparatus for electrochemical analysis of a non-homogeneous sample including:

an electrode, disposed in a solution of the sample to be analyzed;

the shape of the electrode and the disposition of the electrode within the sample being selected such that an output of the electrode relates to the spatial average of a property of the sample.

14. The apparatus of claim 13 wherein:

the property of the sample is the chemical composition of the sample.

15. An apparatus for electrochemical pH-statting of a sample including:

a working electrode disposed in the sample;

a counter electrode;

the working electrode applying a current to the sample to generate in the sample one of the group of ions comprising hydrogen and hydroxyl ions;

the counter electrode being separated from the sample by an electrochemical junction, the counter electrode generating a complementary ion that is separated from the sample by the electrochemical junction so that the pH of the sample is not influenced by the complementary ion;

a pH detector disposed in the sample which detects the pH of the sample; and

,

a controller which regulates the current applied by the working electrode to achieve a preselected pH int he sample.