DEVICE FOR MONITORING AN OXIDATIVE STRESS AND METHODS THEREOF
The present relates, in general terms, to a device for monitoring oxidative stress in a sample, a method of making the device and a method of monitoring oxidative stress in a sample thereof.
1 . A device for monitoring oxidative stress in a sample, comprising:
a) a substrate;
b) a layer coated on the substrate; and
c) a compound having a moiety which is responsive to an oxidative stress marker in the sample, the compound doped within or on the surface of the layer;
wherein the substrate and the layer are optically clear in the wavelength of about 400 nm to about 1000 nm.
2 . The device according to claim 1 , wherein the moiety on the compound is responsive to a carbonyl moiety, and preferably fluorescent
3 . The device according to claim 1 or 2 , wherein the moiety on the compound forms a reversible bond with a carbonyl moiety.
4 . The device according to any of claims 1 to 3 , wherein the moiety on the compound is thiosemicarbazide, or dithianyl.
5 . The device according to any of claims 1 to 4 , wherein the compound is fluorescein-5-thiosemicarbazide (FTSC).
6 . The device according to any of claims 1 to 5 , wherein the compound is doped within layer up to about 10 wt % of the layer, preferably 0.1 wt % to 1 wt % of the layer.
7 . The device according to any of claims 1 to 5 , wherein the compound is doped on the surface of the layer up to about 10 wt % of the layer, preferably 0.1 wt % to 1 wt % of the layer.
8 . The device according to claim 6 or 7 wherein the compound is FTSC and is doped at about 0.1 wt % to about 1 wt % of the layer.
9 . The device according to any of claims 1 to 8 , wherein the layer is a polymer layer, preferably a polymer which comprises a straight chain polymer.
10 . The device according to claim 9 , wherein the polymer layer comprises a polymer selected from acrylate polymer, sulphonated polyetheretherketone, silk, polyacrylamide, vinylimidazole polymer, acrylonitrile butadiene styrene, photopolymer, or copolymers of the above.
11 . The device according to any of claim 9 , wherein the polymer layer comprises acrylated-based photopolymer, and preferably e-shell 300 acrylate-based photpolymer.
12 . The device according to any of claims 1 to 8 , wherein the layer is a glass layer.
13 . The device according to any of claims 1 to 12 , wherein the layer has a thickness of up to 500 μm, preferably up to 100 μm.
14 . The device according to any of claims 1 to 13 , wherein the layer is coated at an end of the substrate.
15 . The device according to any of claims 1 to 14 , wherein the layer coats the entire surface of the substrate.
16 . The device according to any of claims 1 to 15 , wherein an emitted electrical or optical signal from the compound is detectable at an uncoated end of the substrate.
17 . A device for monitoring oxidative stress in a sample, comprising:
a) an optically clear substrate;
b) an optically clear acrylate polymer layer, the polymer layer coated on the substrate; and
c) a fluorescent compound having a thiosemicarbazide moiety which is responsive to a carbonyl moiety in the sample, the fluorescent compound doped within the polymer layer;
wherein the substrate and the polymer layer are optically clear in the wavelength of about 400 nm to about 1000 nm.
18 . A method of making a device for monitoring oxidative stress in a sample, including
a) mixing a monomer with a fluorescent compound to form a mixture, the monomer for forming an optically clear polymer and the fluorescent compound having a moiety which is responsive to an oxidative stress marker in the sample;
b) contacting the mixture with an optically clear substrate; and
c) polymerising the mixture on the substrate for forming a polymer layer coated on the substrate;
wherein the substrate and the polymer layer are optically clear in the wavelength of about 400 nm to about 1000 nm.
19 . The method according to claim 18 , wherein the mixing step comprises vortexing, sonicating or a combination thereof.
20 . The method according to claim 18 or 19 , wherein the monomer is selected from a acrylate-based liquid photo-reactive photomonomer.
21 . The method according to anyone of claims 18 to 19 wherein the fluorescent compound is FTSC.
22 . The method according to any of claims 18 to 21 , wherein the polymerisation step comprises irradiating the mixture with a light at a wavelength of about 300 nm to about 600 nm.
23 . The method according to any of claims 18 to 22 , wherein the polymer layer is coated at an end of the substrate.
24 . The method according to any of claims 18 to 23 , further including connecting an uncoated end of the substrate to a light source and detector for measuring the fluorescence.
25 . A method of monitoring oxidative stress in a sample, including:
a) contacting a device as defined in anyone of claims 1 to 17 with the sample;
b) detecting a fluorescence signal from the device, the fluorescence signal being generated in response to an oxidative stress marker in the sample; and
c) quantifying the fluorescence signal compared to a control signal.
26 . The method according to claim 25 , wherein the response time of the device is up to about 60 sec, for monitoring dynamic changes in oxidative stress in the sample.
27 . The method according to claim 25 or 26 , for use in in-vivo monitoring of oxidative stress and dynamic patterns.
28 . The method according to anyone of claims 25 to 27 for use in intracytoplasmic sperm injection (ICSI) technology, plant cell or animal cell physiological studies, or determining human performance.