Microfluidic device, system and methods thereof for measuring and recording electrical signals from a pool of multiple nematodes

Patent metadata

Field	Value
Publication number	US-2019387717-A1
Application number	US-201916449438-A
Country	US
Kind code	A1
Filing date	Jun 23, 2019
Priority date	Jun 22, 2018
Publication date	Dec 26, 2019
Grant date	—

Abstract

The present disclosure provides a microfluidic device and system for measuring a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes, wherein the composite EPG signal is measured from the pool of nematodes present in a single recording channel connected to two or more integrated electrodes. The microfluidic device includes an inlet port and outlet port directly connected to a single recording channel and two or more electrodes directly connected to the recording channel. The recording channel is configured to hold 10 to 10,000 nematodes.

First claim

1 . A microfluidic device for measuring a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes comprising:
a) an inlet port and outlet port directly connected to a holding reservoir; b) a single recording channel, configured to hold 10 to 10,000 nematodes, connected in series to the holding reservoir; and, c) two or more integrated electrodes directly connected to the recording channel.

2 . The device of claim 1 , wherein the recording channel is 10 mm to 500 mm in length and 10 μm to 500 μm in width.

3 . The device of claim 1 , wherein the device comprises a silicone polymer, a thermoplastic polymer, an acrylic polymer, or a polycarbonate polymer.

4 . (canceled)

5 . The device of claim 3 , wherein the silicone polymer comprises a polydimethylsiloxane (PDMS) elastomer.

6 . The device of claim 1 , wherein the device comprises a backing.

7 - 10 . (canceled)

11 . A microfluidic system for recording a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes comprising:
a) an inlet port and outlet port directly connected to a holding reservoir; b) a single recording channel, configured to hold 10 to 10,000 nematodes, connected in series to the holding reservoir; c) two or more integrated electrodes directly connected to the recording channel; and, d) at least one differential amplifier or at least one voltage-clamp amplifier, wherein the amplifier is connected to an output from the two or more integrated electrodes.

12 - 17 . (canceled)

18 . The system of claim 11 , wherein the differential amplifier or voltage-clamp amplifier is connected to the two or more integrated electrodes via a recording dock.

19 . A method for recording a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes comprising:
a) introducing the pool of multiple nematodes into the holding reservoir through the inlet port of the microfluidic system of claim 11 , wherein the pool of multiple nematodes is present in an aqueous buffer solution; b) moving the pool of multiple nematodes into the single recording channel; c) measuring electrophysiological signals from the pool of multiple nematodes; and, d) recording the electrophysiological signals as a single composite EPG.

20 . The method of claim 19 , wherein the buffer solution comprises serotonin, food, or other stimulant to cause pharyngeal pumping.

21 . (canceled)

22 . (canceled)

23 . The method of claim 19 , wherein the pool of nematodes comprises transgenic nematodes or transgenic nematodes comprising a gene variant.

24 . (canceled)

25 . (canceled)

26 . The method of claim 19 , wherein the moving the pool of multiple nematodes into the single recording channel comprises applying a vacuum, positive pressure, or centrifugal force to the system.

27 . A method for screening test compounds by recording a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes using the microfluidic system of claim 11 , comprising:
a) contacting the pool of multiple nematodes with the test compound; b) measuring and recording a composite EPG from the contacted pool of multiple nematodes; c) comparing the recorded composite EPG to a control composite EPG; and, d) determining if the recorded composite EPG is altered as compared to the control composite EPG, whereby test compounds are screened.

28 . The method of claim 27 , wherein determining if the recorded composite EPG is altered comprises determining the power spectrum of the composite EPG, the frequency of the peak power of the composite EPG, the amplitude of the composite EPG, waveform of the composite EPG, or a combination thereof.

29 . (canceled)

30 . (canceled)

31 . (canceled)

32 . The method of claim 27 , wherein the pool of nematodes comprises transgenic or variant nematodes.

33 . (canceled)

34 . (canceled)

35 . The method of claim 27 , wherein the test compound is selected from a drug, a drug candidate, an industrial chemical, or an environmental pollutant.

36 . The method of claim 35 , wherein the drug or drug candidate is selected from an organic compound, an inorganic compound, a hormone, a growth factor, a cytokine, a receptor, an antibody, an enzyme, a peptide, an aptamer or a vaccine.

37 . A microfluidic device for measuring a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes comprising:
a) an inlet port and outlet port directly connected to a single recording channel; b) the single recording channel, configured to hold 10 to 10,000 nematodes; c) a tube placed in each of the inlet port and outlet port, wherein the tube placed in the outlet port comprises a filter to retain nematodes in the single recording channel; and, c) two or more electrodes connected to the recording channel.

38 - 48 . (canceled)

49 . A microfluidic system for recording a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes comprising the microfluidic device of claim 37 , the system comprising:
a) an inlet port and outlet port directly connected to a single recording channel; b) the single recording channel, configured to hold 10 to 10,000 nematodes; c) a tube placed in each of the inlet port and outlet port, wherein the tube placed in the outlet port comprises a filter to retain nematodes in the single recording channel; d) two or more electrodes connected to the recording channel; and, e) at least one differential amplifier or at least one voltage-clamp amplifier, wherein the amplifier is connected to an output from the two or more electrodes.

50 - 56 . (canceled)

57 . A method for recording a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes comprising:
a) introducing the pool of multiple nematodes into the single recording channel through the inlet port of the microfluidic system of claim 49 , wherein the pool of multiple nematodes is present in an aqueous buffer solution; b) measuring electrophysiological signals from the pool of multiple nematodes; and, c) recording the electrophysiological signals as a single composite EPG.

58 - 65 . (canceled)

58 . A method for screening test compounds by recording a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes using the microfluidic system of claim 49 , the method comprising:
a) contacting the pool of multiple nematodes with a first buffer; b) measuring and recording a composite EPG from the first buffer pool of multiple nematodes; c) perfusing the pool of multiple nematodes with a second buffer; d) measuring and recording a composite EPG from the second buffer pool of multiple nematodes; e) comparing the recorded composite EPG from the first buffer pool of multiple nematodes to the recorded composite EPG from the second buffer pool of multiple nematodes; and, d) determining if the recorded composite EPG from the first buffer pool of multiple nematodes is altered as compared to the recorded composite EPG from the second buffer pool of multiple nematodes, whereby test compounds are screened,
wherein either of the first buffer or second buffer comprises a test compound and the other buffer is a control buffer.

67 - 77 . (c

Assignees

• Nemametrix Inc
• Univ Oregon

Inventors

• Lockery Shawn

Classifications

• G01N33/5085

  of invertebrates · CPC title

• G01N2333/4353

  from nematodes · CPC title

• A01K29/005 Primary

  Monitoring or measuring activity · CPC title

• A61B2503/40

  Animals · CPC title

• A61B2503/42

  for laboratory research · CPC title