Device for online measurement of neurotransmitters using enzymatic reactors

• US 9,879,305 B2
• Filed: 05/02/2012
• Issued: 01/30/2018
• Est. Priority Date: 05/02/2012
• Status: Active Grant

• Current Assignee: Alberto Morales Villagran, Daniel Morales Rodriguez
• Original Assignee: Alberto Morales Villagran, Daniel Morales Rodriguez
• Inventors: Morales Villagran, Alberto, Morales Rodriguez, Daniel
• Primary Examiner(s): Bowers, Nathan A
• Application Number:
  US14/398,426
• Publication Number:
  US 20150140586A1
• Time in Patent Office: 2,099 Days
• CPC Class Codes: C12Q 1/44 involving esterase
• CPC Class Codes: C12Q 1/52 involving transaminase
• CPC Class Codes: G01N 33/9406 Neurotransmitters

Abstract

The invention relates to a device and a method for measuring neurotransmitters in the brain dialysate of mammals using enzymatic reactors specific to a certain neurotransmitter, which generate fluorescent derivatives when mixed. The mixture is introduced into an incubation chamber and passes through a temperature controlling system in order to set it to the optimum enzymatic reaction temperature, and the fluorescent derivative generated is passed through a flow fluorescence cell located inside the incubation chamber, where the liquid is impacted with a laser light source via an optical fiber. The fluorescence emitted is captured by a fluorescence detector that sends it to a processing system in order to obtain the corresponding values.

Claims

1. A device for online measurement of neurotransmitters comprising:

an incubation chamber made of a light transparent material, the incubation chamber including inside;

a fluorescent cell having a first inlet, a second inlet, a first outlet, and a second outlet, wherein the second outlet conduit is perpendicular to the second inlet conduit;

a silica tube having a first end connected to the first inlet of the fluorescent cell;

a Y-connector connected to a second end of the silica tube, the Y-connector having a first end and a second end; and

an adjustable temperature mat located on a bottom of the incubation chamber;

a pump having a first pipe in fluid flow communication with the pump adapted to transport a brain dialysate to the first end of the Y-connector inside the incubation chamber and a second pipe in fluid flow communication with the pump adapted to transport a selected enzymatic reactant to the second end of the Y-connector inside the incubation chamber, wherein the brain dialysate and the selected enzymatic reactant converge simultaneously in the Y-connector forming a mixture;

a laser light source connected to the second inlet of the fluorescent cell by using an optical fiber, the laser light source emits a laser light for excitation of the mixture flowing through the fluorescent cell;

a heating device connected to the mat to regulate the temperature of the mat;

a fluorescence detecting and measuring device connected to the second outlet of the fluorescent cell through optical fiber for detection and measurement of fluorescent light emitted by the mixture subjected to excitation; and

wherein when the mixture passes through the silica tube placed inside the incubation chamber, the mat heats the mixture to an optimal temperature for the enzymatic reactant.

2. The device of claim 1, wherein the silica tube is made of a fused silica tube.

3. The device of claim 1, wherein the incubation chamber is about 0.8 mm per side and 1.5 mm long (0.96 microliters).

4. The device of claim 1, wherein the optical fibers have a diameter of 600 microns.

5. The device of claim 1, wherein the heating device comprises a temperature control by induction of voltage pulses programmed in a microprocessor to achieve a resolution of ±

O.PC.

6. The device of claim 1, wherein the heating device further comprises a temperature controlled oven including perforations for connection to the mat.

7. The device of claim 1, wherein the fluorescence detecting and measuring device is a CCD detector and/or a photomultiplier.

8. The device of claim 1, wherein the first pipe and the second pipe of the pump have a capillary of 0.8 mm square and 1.5 mm straight section.

9. The device of claim 1, wherein the device analyzes online fluorescent derivatives with a resolution less than one second.

10. The device of claim 1, wherein the fluorescent cell has an internal volume of about 1 microliter at the area of fluorescence excitation and emission.