WO2018031675A1 - Développement d'un réseau de micro-électrodes à fibre de carbone multicanal pour mesures électrochimiques - Google Patents

Développement d'un réseau de micro-électrodes à fibre de carbone multicanal pour mesures électrochimiques Download PDF

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Publication number
WO2018031675A1
WO2018031675A1 PCT/US2017/046126 US2017046126W WO2018031675A1 WO 2018031675 A1 WO2018031675 A1 WO 2018031675A1 US 2017046126 W US2017046126 W US 2017046126W WO 2018031675 A1 WO2018031675 A1 WO 2018031675A1
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WO
WIPO (PCT)
Prior art keywords
array
carbon fiber
microelectrode
gold
lower portion
Prior art date
Application number
PCT/US2017/046126
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English (en)
Inventor
Michael L. HEIEN
James R. SIEGENTHALER
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Arizona Board Of Regents On Behalf Of The University Of Arizona
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Application filed by Arizona Board Of Regents On Behalf Of The University Of Arizona filed Critical Arizona Board Of Regents On Behalf Of The University Of Arizona
Publication of WO2018031675A1 publication Critical patent/WO2018031675A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/14546—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue for measuring analytes not otherwise provided for, e.g. ions, cytochromes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/1468—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using chemical or electrochemical methods, e.g. by polarographic means
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
    • A61B5/6846—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive
    • A61B5/6867—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive specially adapted to be attached or implanted in a specific body part
    • A61B5/6868—Brain
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B2562/00—Details of sensors; Constructional details of sensor housings or probes; Accessories for sensors
    • A61B2562/04—Arrangements of multiple sensors of the same type
    • A61B2562/046—Arrangements of multiple sensors of the same type in a matrix array
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B2562/00—Details of sensors; Constructional details of sensor housings or probes; Accessories for sensors
    • A61B2562/12—Manufacturing methods specially adapted for producing sensors for in-vivo measurements
    • A61B2562/125—Manufacturing methods specially adapted for producing sensors for in-vivo measurements characterised by the manufacture of electrodes

Definitions

  • the present invention relates to detection and measurement of an electroacfive chemical located in the brain of an animal using Fast Scan Cyclic Voltammetry or Fast- scan Controlled Absorption Voltammetry.
  • Tonic levels of dopamine can be measured with single carbon fiber microelectrodes (or alternately, CF Es) using fast-scan cyclic voltammetry ("FSCV”) and fast-scan controiled-absorption voltammetry (“FSCAV").
  • FSCV fast-scan cyclic voltammetry
  • FSCAV fast-scan controiled-absorption voltammetry
  • a major limitation to these measurement techniques is the regional localization as each electrode only samples from 10 microns around its environment based on the microeiectrode placement.
  • a multi-channel carbon fiber microeiectrode array has been developed for both FSCV and FSCAV measurements.
  • the array features a 2x8 microeiectrode placement with 300 micron spacing between fibers, providing the ability to span regions of the brain. Characterization of the array includes single channel analysis by slow scan voltammetry and FSCV.
  • the present invention features a multi-channel CFME array effective for optimizing an acquisition of electrochemical measurements via FSCV or FSCAV.
  • the array may comprise an array structure having a plurality of parallel, equally-sized, and equally-spaced gold traces.
  • the plurality of gold traces and a plurality of insulating barriers are arranged in an alternating pattern in the array structure such that each insulating barrier and each gold trace alternate with each other.
  • a plurality of CFMEs each having an upper portion and a lower portion, also comprises the array.
  • the upper portion of each CFME contacts one of the gold traces, while the lower portion of each CFME extends beyond a bottom of the array structure. Additional embodiments may feature an adhesive affixing each CFME to a gold trace.
  • the upper portion of each CFME and each gold trace has a conductive coating disposed thereon.
  • each CFME has a conductive tip.
  • the lower portion of each CFME, excluding the conductive tip, may be insulated with an insulation material.
  • an insulating seal overlays the entire array structure.
  • This insulating seal along with the plurality of insulating barriers, provides a double layer of insulation to the plurality of CFMEs. In this way, the overall array is effectively insulated while maintaining an isolated eiectroactive area at the end of each carbon fiber microeiecfrode.
  • each CFME may serve as a channel for acquiring electrochemical measurements.
  • the electrochemical measurements are acquired by applying a voltage to each CFME and measuring the resultant current.
  • the array is coupled to a custom amplifier for amplifying the resultant current before measurements are taken. Further, as a result of having multiple channels, the array optimizes the acquisition of the electrochemical measurements, as compared to single CFMEs, by obtaining simultaneous measurements from a plurality of regions of, for example, a test solution or an animal brain.
  • One of the unique and inventive technical features of the present invention is the use of multiple channels in the microelectrode array, which allow for the acquisition of multiple electrochemical measurements that span several regions of, for example, an animal brain.
  • Presently known prior references and work limit the acquisition of electrochemical measurements to single carbon-fiber microeiectrodes due to the problems associated with fabricating and insulating multi-channel carbon-fiber microelectrode arrays.
  • Multi-channel arrays using CFMEs have been developed for acquiring measurements in other applications, but are not compatible with FSCV and FSCAV measurements because of the resistance and capacitance of the arrays.
  • the resistance and capacitance are directly related to the effective insulation of the carbon fibers; thus, the multi-channel array must effectively insulate the carbon fibers while providing a defined eiectroactive area at the end of each carbon fiber.
  • the present invention selectively insulated the carbon fibers, leaving an exposed eiectroactive area at each end.
  • a removable masking material was placed on the end of each carbon fiber prior to insulating the present multi-channel array. After the array was insulated, using electrodeposition, the mask was removed with a compatible solvent (i.e., that would not dissolve any of the other dried epoxies or paint already on the array).
  • FIGs. 1A-1 E are a diagrammatic detailing of the steps to making the multichannel CFME array.
  • F!G. 1 A shows an empty array structure with a plurality of gold traces.
  • FIG. 1 B shows the array structure with torr Seal Epoxy insulating barriers added between the gold traces.
  • FIG. 1 C shows CFMEs tacked into the gold traces using 2-ton epoxy.
  • FIG. 1 D shows a conductive silver paint applied to the CFMEs for increased conductivity of cells.
  • FIG. 1 E shows the array structure overlaid with an insulating 2-ton epoxy.
  • FIG. 2 shows a diagrammatic representation of the multi-channel CFME array
  • FIG. 3 shows an alternate diagrammatic representation detailing of the steps to making the multichannel CFME array.
  • FIG. 4A shows an FSCV calibration curve constructed for dopamine (top left, bottom left).
  • FIG. 4B shows the flow profile for measured dopamine (top right, bottom right).
  • FIG. 5 shows that the selective insulation of a carbon fiber was achieved by masking the carbon fiber with polypropylene followed by an electrodeposited insulating layer. The mask was then removed exposing a defined electroactive area on the carbon fiber.
  • FIG. 6A shows the insulated carbon fiber was cut and imaged using SEM to reveal the insulating film of ClearClad HSR® measured to be approximately 1 -1 .2 m in thickness.
  • FIG. 6B shows the SEM imaging of a carbon fiber that had been selectively insulated as the insulating material.
  • FIG. 7A shows a calibration plot for dopamine (alternately, "DA").
  • FIG. 7B shows selected flow profiles of 1000nM dopamine for each carbon fiber.
  • FIG. 7C shows flow profile of dopamine for two carbon fibers.
  • FIG. 8A shows a calibration curve for each microeiectrode of the dual electrode system.
  • FIG. 8B shows the flow profiles for each microeiectrode of the dual electrode system.
  • the present invention features a multi-channel carbon fiber microeiectrode array (100) effective for optimizing an acquisition of electrochemical measurements via fast-scan cyclic sculptureammetry or fast-scan absorption lakeammetry.
  • the array (100) may comprise an array structure (101 ) having a plurality of parallel, equally-sized, and equally-spaced gold traces (102a, ... , 102n).
  • the center-to-center spacing between gold traces is 300 microns.
  • each gold trace is disposed between two insulating barriers as can be seen in FIGs. 1 B-1 E and F!G. 2.
  • the number of gold traces can be greater than, or alternatively, less than the number of insulating barriers.
  • the number of gold traces varies from about 2 to about 18, In another embodiment, the number of gold traces is at least about 2.
  • the number of insulating barriers range from about 2 to about 14. The number of insulating barriers may be at least 2. However, since an insulating barrier is disposed between each electrode, the number of insulating barriers depend on the number of carbon fiber electrodes in the array. Moreover, it can be appreciated that any number of carbon fiber electrodes can be included in the array.
  • the plurality of insulating barriers (103a, ..., 103m) are composed of torr seal epoxy.
  • the number of carbon fiber microeiectrodes may be, at most, equal to the number of gold traces.
  • a variety of ratios of the length of the upper portion to the length of the lower portion of the carbon fiber microeiecfrode may be employed, where the ratio is restricted only by the requirement that the upper portion is in electrical contact with a gold trace.
  • Non-limiting examples of the ratio between the length of the upper portion to the length of the lower portion include 50:50, or 75:25, or 25:75, etc.
  • each carbon fiber microelectrode contacts one of the gold traces, while the lower portion of each carbon fiber microelectrode extends beyond a bottom of the array structure (101 ).
  • Additional embodiments may feature an adhesive (105a, ...,105n) (e.g., 2-ton epoxy) affixing each carbon fiber microelectrode to a gold trace.
  • the upper portion of each carbon fiber microelectrode and each gold trace has a conductive coating (106a, ...,106n) disposed thereon.
  • the conductive coating is either conductive colloidal silver paint or conductive carbon paint.
  • each carbon fiber microelectrode has a conductive tip.
  • the lower portion of each carbon fiber microelectrode, excluding the conductive tip, may be insulated with an insulation material.
  • the insulation material is an insulative polymer or a silica capillary.
  • a predetermined length of the lower portion of each carbon fiber microelectrode may be (i.e., cut).
  • an insulating seal (107) overlays the entire array structure (101 ). This insulating sea! (107), along with the plurality of insulating barriers (103a,... ,103m), provides a double layer of insulation to the plurality of carbon fiber microelectrodes (104a, ... , 104n).
  • the overall array is effectively insulated while defining an isolated electroactive area for each carbon fiber microelectrode for acquisition of said electrochemical measurements.
  • the lower portion of each carbon fiber microelectrode is the isolated electroactive area serving as a channel for acquiring eiectrochemical measurements.
  • the array (100) optimizes the acquisition of electrochemical measurements, as compared to single carbon fiber microelectrodes, by obtaining simultaneous measurements from a plurality of regions of a brain.
  • the array (100) may be employed for fast- scan cyclic voltammetry or fast-scan absorption voltammetry with an average scan rate of 400 volts per second and a total current of less than 2000 microamps.
  • This configuration may yield an overall array resistance between 50 Ohms and 100 Ohms and a maximum collective capacitance of 5 nano Farads.
  • Fabrication of the multi-channel carbon fiber microelectrode array of the present invention was done by hand under a microscope. Initial designs of the array included eight gold traces spaced 300pm apart, center-to-center. The total distance across the array was 2.61 mm.
  • Tools necessary for the construction of the array include a surgical scalple, pulled glass capiilarys, and microforcepts.
  • a barrier of Hysoi 2-part epoxy was applied between each gold trace, forming a barrier between the traces. The epoxy was allowed to dry overnight.
  • a carbon fiber with a length between 1500 ⁇ and 2500 ⁇ was paced onto each gold trace and tacked in place by placing a small drop of epoxy at the end of the PCB using clear 2-Ton epoxy applied wih a pulled glass capillary. The epoxy was allowed to harden fully by placing the array in an oven at 1 15°C for 2hrs.
  • each gold trace and carbon fiber was made by painting the carbon fibers and gold traces with either conductive collodiai silver paint or conductive carbon paint by immersing the paint in acetone or isopronyl, aspirating a pulled capillary with the paint, and dispensing the paint using capillary action.
  • the electrically connected carbon fibers were then insulated by painting the connection with 2-ton epoxy, which was allowed to dry either overnight, or by placement in an oven at 1 15°C for 2hrs.
  • the carbon fibers that exteneded from the epoxy edge were then trimmed to a length of 1000 ⁇ from the epoxy edge.
  • An alternative method to placing raw carbon fibers and electricially insulating them to the desired length is to place preinsulated carbon fibers in place. This can be done by modifying the method of Phillips et. ai. Breifley, carbon fibers are aspirated while submerged in isoprolyl alcohol into a silica capillary that is 1500 ⁇ long, and having an outer diameter of 90 ⁇ . The aspirated capillaries are removed from the solution and allowed to dry. A small bead of epoxy is placed on the carbon fiber, and the carbon fiber is gently pulled through the capillary allowing the epoxy to wick inside the capiiary. A second bead is placed and the carbon fiber is further pulled. The bead is pulled until!
  • the capillary is allowed to dry.
  • the carbon fiber extending from the dired epoxy is then trimmed to length, typically 50-100 ⁇ .
  • the aspirated capillary is placed on the PCB and tacked in place. The electrical connection process and insulation procedures remain unaltered.
  • the array was tested to determine if an electrical connection was made between the gold traces and carbon fibers by placing the array in a solution of artificial cerebral spinal fluid and cycling the array from -0.4 to 1.3V at 400V/s repeated at 60Hz. Upon successful determination, a single carbon fiber on the PCB was trimmed to be 100 ⁇ in length from the epoxy edge and was calibrated to determine the linearity of the response (see FIGs. 3A-3B.
  • the flow-cell color plot (FIG. 5B) and calibration curve (FIG. 4A) demonstrates that the array is capable of quantitativly measuring dopamine with a linear response from 50nM to 1000nM.
  • the exposed ⁇ ⁇ carbon fibers must be reduced to have an eiectroactive area of a tunable size between 50-100 ⁇ .
  • Insulation of the carbon fibers was achieved by using a commercially available electro-depositable paint, CiearClad HSR ⁇ using modified deposition parameters from Sripirom, et. al.
  • CiearClad HSR ⁇ is a polyurethane suspension that is applied by cathodic electrodeposition, creating a solvent and electrically resistive insulating film. The thickness can be tuned based on applied deposition voltage and time.
  • the tip of each carbon fiber were masked with melted polypropylene ("PP").
  • Electric deposition of the film was conducted by using chronoamperometry in a 2-electrode setup with a silver wire as the counter electrode. The coating was applied in two coats, the first was applied at a -4V potential for 120 seconds. The carbon fiber was then briefly rinsed in deionized water and heat cured in an oven at 1 15°C for 20 minutes. A second coating was completed by applying -8V for 120 seconds followed by a brief rinse and heat cure at 1 15°C. The array was then suspended in toluene and sonicated for 10 minutes to remove the PP mask (FSG. 4).
  • the above parameters deposit an insulating layer of ⁇ 1 ⁇ thickness when a cross section of each coated carbon fiber is cut and imaged using scanning electron microscope (“SEM”) imaging (FIGs. 6A-8B). SEM also shows that each carbon fiber is selectively insulated from where the mask was placed and removed indicating a defined electroactive area that is reduced from the original size.
  • SEM scanning electron microscope
  • the array was placed in a modified flow- cell setup. Artificial cerebral spinal fluid (“ACSF”) was flowed across the array, and dopamine was injected as a bolus into the flow path.
  • ACSF Artificial cerebral spinal fluid
  • a homebui!t duai-e!ectrode potentiostat was utilized to measure two carbon fibers simultaneously for the detection of dopamine.
  • a linear calibration curve was generated and solution flow dynamics were probed utilizing the array. Simultaneous detection of dopamine on multiple fibers was determined to be possible.
  • FIG. 8A shows a calibration curve for each microelectrode and FIG. 8B shows the flow profiles for each microelectrode.
  • references to the inventions described herein using the phrase “comprising” includes embodiments that could be described as “consisting of, and as such the written description requirement for claiming one or more embodiments of the present invention using the phrase “consisting of is met.

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Abstract

Les niveaux toniques de dopamine peuvent être mesurés à l'aide de micro-électrodes à fibre de carbone unique à l'aide de la voltampérométrie cyclique à balayage rapide ("FSCV") et de la voltampérométrie à absorption contrôlée à balayage rapide ("FSCAV"). Une limitation majeure de ces techniques de mesure est la localisation régionale sur la base du placement de la micro-électrode. Dans un effort pour surmonter ce défi, un réseau de micro-électrodes à fibre de carbone multicanal a été développé pour des mesures FSCV et FSCAV. Le réseau comprend un placement de 2x8 micro-électrode avec un espacement de 300 micromètres entre les fibres, ce qui permet de couvrir les régions du cerveau. La caractérisation du réseau comprend une analyse de canal unique par voltampérométrie lente et FSCAV.
PCT/US2017/046126 2016-08-09 2017-08-09 Développement d'un réseau de micro-électrodes à fibre de carbone multicanal pour mesures électrochimiques WO2018031675A1 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US201662372652P 2016-08-09 2016-08-09
US62/372,652 2016-08-09
US201662373292P 2016-08-10 2016-08-10
US62/373,292 2016-08-10
US201762508942P 2017-05-19 2017-05-19
US62/508,942 2017-05-19

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050230270A1 (en) * 2002-04-29 2005-10-20 The Trustees Of Boston College And Battelle Memorial Institute Carbon nanotube nanoelectrode arrays
WO2010103174A1 (fr) * 2009-03-09 2010-09-16 Oulun Yliopisto Électrode multicanaux en fibres de carbone utilisée pour mesurer l'activité électrique et chimique dans un tissu biologique et procédé de fabrication associé
US20140342128A1 (en) * 2011-10-14 2014-11-20 Digital Sensing Limited Arrays and methods of manufacture
US20150250421A1 (en) * 2012-09-26 2015-09-10 Advanced Diamond Technologies, Inc. Conductive nanocrystalline diamond micro-electrode sensors and arrays for in-vivo chemical sensing of neurotransmitters and neuroactive substances and method of fabrication thereof
US20170007824A1 (en) * 2013-07-05 2017-01-12 Trustees Of Boston University Minimally invasive splaying microfiber electrode array and methods of fabricating and implanting the same

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050230270A1 (en) * 2002-04-29 2005-10-20 The Trustees Of Boston College And Battelle Memorial Institute Carbon nanotube nanoelectrode arrays
WO2010103174A1 (fr) * 2009-03-09 2010-09-16 Oulun Yliopisto Électrode multicanaux en fibres de carbone utilisée pour mesurer l'activité électrique et chimique dans un tissu biologique et procédé de fabrication associé
US20140342128A1 (en) * 2011-10-14 2014-11-20 Digital Sensing Limited Arrays and methods of manufacture
US20150250421A1 (en) * 2012-09-26 2015-09-10 Advanced Diamond Technologies, Inc. Conductive nanocrystalline diamond micro-electrode sensors and arrays for in-vivo chemical sensing of neurotransmitters and neuroactive substances and method of fabrication thereof
US20170007824A1 (en) * 2013-07-05 2017-01-12 Trustees Of Boston University Minimally invasive splaying microfiber electrode array and methods of fabricating and implanting the same

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZACHEK ET AL.: "Electrochemical dopamine detection: Comparing gold and carbon fiber microelectrodes using background subtracted fast scan cyclic voltammetry", JOURNAL OF ELECTROANALYTICAL CHEMISTRY, vol. 614, 2008, pages 113 - 120, XP022501428 *

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