Arrangement and method using microsensors for measuring cell vitalities

• US 8,916,035 B2
• Filed: 09/29/2010
• Issued: 12/23/2014
• Est. Priority Date: 09/30/2009
• Status: Active Grant

• Current Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Original Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Inventors: Frey, Alexander, Gumbrecht, Walter, Paulicka, Peter, Schienle, Meinrad, Stanzel, Manfred, Sickert, Daniel
• Primary Examiner(s): Brown, Melanie Y
• Assistant Examiner(s): MARTINEZ, REBECCA L
• Application Number:
  US13/499,376
• Publication Number:
  US 20120252055A1
• Time in Patent Office: 1,546 Days
• Field of Search: None
• US Class Current: 204/403.01
• CPC Class Codes: C12M 41/38 of metabolites or enzymes in the cells
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: G01N 27/404 Cells with anode, cathode and cell electrolyte on the same side of a permeable membrane which separates them from the sample fluid , e.g. Clark-type oxygen sensors
• CPC Class Codes: G01N 27/416 Systems G01N27/27 takes precedence
• CPC Class Codes: G01N 27/423 Coulometry
• CPC Class Codes: G01N 33/48735 Investigating suspensions of cells, e.g. measuring microbe concentration by chemical means C12Q1/04; colony counters C12M1/34; concentration of particle suspensions in general G01N15/06

Abstract

An arrangement and a method measures cell vitalities with a sensor array. The sensor array is formed on a surface of a semiconductor chip. The semiconductor chip has integrated circuits and an integrated circuit is associated with each sensor of the sensor array, for processing the measurement signals of the respective sensor. The integrated circuits are formed in the semiconductor chip spatially in each case below the associated sensor and neighboring sensors of the sensor array have a center-to-center in the range of micrometers. The pH and/or pO₂ can be measured in the environment of a living cell.

Claims

1. A device to measure cell vitalities, comprising:

a semiconductor chip;

a sensor array formed on one surface of the semiconductor chip, the sensor array comprising a plurality of sensors, each sensor producing a measurement signal and comprising a working electrode, the sensor array being arranged such that midpoints of adjacent sensors are separated by a distance of from 1 to 999 micrometer; and

integrated circuits formed in the semiconductor chip such that an integrated circuit is assigned to each sensor of the sensor array to process the measurement signal produced by the assigned sensor, the integrated circuits in the semiconductor chip are each spatially formed under the sensor assigned thereto, and each integrated circuit comprises

a first switching transistor which supplies the working electrode of the assigned sensor with a first voltage when the first switching transistor is closed,

a second switching transistor which supplies the working electrode of the assigned sensor with a second voltage, different from the first voltage, when the second switching transistor is closed,

a voltage follower transistor that provides a third voltage that follows a voltage supplied to the voltage follower transistor by the working electrode of the assigned sensor, and

a selection transistor which, together with the voltage follower transistor, electrically selects the assigned sensor by a column and a row of the sensor array.

2. The device as claimed in claim 1, wherein the midpoints of adjacent sensors are separated by a distance in a range of from 1 to 100 micrometers.

3. The device as claimed in claim 1, wherein the midpoints of adjacent sensors are separated by a distance in a range of from 1 to 10 micrometers.

4. The device as claimed in claim 1, wherein

the sensors are amperometric or coulometric electrochemical sensors, and/or

the sensors each comprise at least one interdigital electrode as a working electrode.

5. The device as claimed in claim 1, wherein each integrated circuit comprises a correlated double sampling stage (CDS) to suppress electrical noise signals and suppress electrical drift of the assigned sensor, and the correlated double sampling stage (CDS) is arranged under the assigned sensor or in a region of the semiconductor chip outside of the sensor array.

6. The device as claimed in claim 1, further comprising a temperature controller to set a defined temperature over the sensor array.

7. The device as claimed in claim 1, wherein further comprising means for immobilizing living cells over the sensors of the sensor array.

8. The device as claimed in claim 7, wherein the means for immobilizing living cells comprises a filter membrane arranged in fluid contact with the sensors.

9. The device as claimed in claim 7, wherein

the means for immobilizing living cells comprises a magnetic field-creating device, which is configured to create a magnetic field over the sensors of the sensor array, and

the living cells are bound to magnetic particles.

10. The device as claimed in claim 9, wherein the magnetic field-creating device immobilizes the living cells over the sensor array in the form of an essentially uniformly thick matrix of magnetic particles with the living cells embedded in the matrix of magnetic particles.

11. The device as claimed in claim 10, wherein

the magnetic field is switchable between an on state and an off state, and

dead or damaged cells are removed in the off state of the magnetic field.

12. The device as claimed in claim 1, wherein

the semiconductor chip is surrounded by a flow cell, and

the sensors of the sensor array are arranged in fluid contact with a flow channel of the flow cell.

15. The device as claimed in claim 1, wherein the voltage follower transistor has a gate terminal electrically connected to the working electrode.

16. The device as claimed in claim 1, wherein when the selection transistor is closed, the third voltage is supplied through the selection transistor.

17. The device as claimed in claim 1, wherein the second switching transistor supplies the working electrode of the assigned sensor with the second voltage when the second transistor is closed while the first switching transistor is open.

13. A method for measuring cell vitalities, comprising:

providing a device comprising;

a semiconductor chip;

a sensor array formed on one surface of the semiconductor chip, the sensor array comprising a plurality of sensors, each sensor producing a measurement signal and comprising a working electrode, the sensor array being arranged such that midpoints of adjacent sensors are separated by a distance of from 1 to 999 micrometer; and

integrated circuits formed in the semiconductor chip such that an integrated circuit is assigned to each sensor of the sensor array to process the measurement signal produced by the assigned sensor, the integrated circuits in the semiconductor chip are each spatially formed under the sensor assigned thereto, and each integrated circuit comprises

a first switching transistor which supplies the working electrode of the assigned sensor with a first voltage when the first switching transistor is closed,

a second switching transistor which supplies the working electrode of the assigned sensor with a second voltage, different from the first voltage, when the second switching transistor is closed,

a voltage follower transistor that provides a third voltage that follows a voltage supplied to the voltage follower transistor by the working electrode of the assigned sensor, and

a selection transistor which, together with the voltage follower transistor, electrically selects the assigned sensor by a column and a row of the sensor array;

bringing living cells into contact with the sensors; and

measuring pH and/or the pO₂ in a vicinity of the living cells using the sensors of the sensor array.

14. The method as claimed in claim 13, further comprising:

setting a temperature over the sensor array to 37°

C. or a different temperature optimal for cell vitality; and

/or

supplying nutrients and/or oxygen to the cells which favor cell vitality; and

/or

supplying antibiotics or other substances to the cells to impair cell vitality.