Methods and systems for transmission and detection of free radicals
The present disclosure provides systems and methods for characterizing the interaction of free radicals with various materials and the use of known interactions to isolate free radical generation from free radical interaction with a target molecule.
1. A system comprising:
a free radical generation chamber;
a free radical source positioned within the free radical generation chamber, the free radical source configured to generate one or more free radicals;
an interaction chamber; and
a free radical transmitting barrier positioned between the free radical generation chamber and the interaction chamber, the free radical transmitting barrier comprising a barrier material having a known response to interaction with the one or more free radicals.
2. A method comprising:
a) generating a plurality of free radicals from a free radical source, the free radical source positioned on a first side of a material of interest that is opposite a second side of the material of interest, wherein the plurality of free radicals is moving in a direction toward the material of interest;
b) measuring a measurable change in at least one property of a free radical detection species positioned on the second side of the material of interest, the measuring occurring after a length of time where at least a portion of the plurality of free radicals have passed through the material of interest and interacted with the free radical detection species, the free radical detection species undergoing the measurable change in at least one property after interaction with one or more of the plurality of free radicals.
3. A method of modifying a target molecule located in a sample, the sample located in an interaction chamber positioned on a second side of a free radical transmitting barrier, the method comprising:
a) generating a plurality of free radicals from a free radical source, the free radical source positioned in a free radical generation chamber positioned on a first side of the free radical transmitting barrier that is opposite the second side, wherein the plurality of free radicals is moving in a direction toward the free radical transmitting barrier and the interaction chamber; and
b) waiting a length of time sufficient for one or more of the plurality of free radicals to pass through the free radical transmission barrier and interact with the target molecule, thereby modifying the target molecule.
4. The method of claim 3 , wherein the generating a plurality of free radicals of step a) includes generating a plasma.
5. The method of claim 4 , wherein the generating a plasma includes generating a sequence of plasma pulses.
6. The method of claim 5 , wherein the sequence of plasma pulses has a pulse width of between 1 ps and 1 ms.
7. The method of claim 5 , wherein the sequence of plasma pulses has a frequency of between 1 Hz and 100 GHz.
8. The method of claim 5 , wherein the sequence of plasma pulses is generated for a total length of time of between 1 ns and 1 hour.
9. The method of claim 4 , wherein the plasma of step a) is generated by a voltage in a range of 1 V and 1 MV.
10. The method of claim 3 , wherein the generating of step a) is configured to provide a peak concentration of marker radicals in the sample in a range of between 50 nM and 800 μM.
11. The method of claim 3 , wherein the sample has a volume of between 1 μL and 400 L.
12. The method of claim 3 , wherein the free radical generation chamber includes a fluid containing a plurality of marker radical precursors.
13. The method of claim 3 , wherein the sample is selected from the group consisting of blood, blood plasma, urine, saliva, lymph, tears, sweat, cerebrospinal fluid, amniotic fluid, aqueous humour, vitreous humour, bile, breast milk, cerumen, chyle, chime, endolymph, perilymph, exudates, feces, female ejaculate, gastric acid, gastric juice, mucus, pericardial fluid, peritoneal fluid, pleural fluid, pus, rheum, sebum, serious fluid, semen, smegma, sputum, synovial fluid, vaginal secretion, vomit, living bacterial cultures, living tissue or eukaryotic cell cultures, and combinations thereof.
14. The method of claim 3 , wherein the sample is selected from the group consisting of eukaryotic intracellular fluid, eukaryotic extracellular fluid, prokaryotic intracellular fluid, prokaryotic extracellular fluid, homogenized tissue or cells, homogenized tissue or cell culture, homogenized plant tissue, and combinations thereof.
15. The method of claim 14 , wherein the extracellular fluid is selected from the group consisting of intravascular fluid, interstitial fluid, lymphatic fluid, transcellular fluid, plant apoplastic or vascular fluid, excess nutrient medium from prokaryotic or eukaryotic in vitro growth, and combinations thereof.
16. The method of claim 3 , wherein the sample comprises the target molecule and a buffer solution.
17. The method of claim 16 , wherein the buffer solution comprises phosphate buffered saline, tris (hydroxymethyl) aminomethane (tris), tris hydrochloric acid, ammonium bicarbonate, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 3-(Nmorpholino) propanesulfonic acid (MOPS), 2-(N-morpholino) ethanesulfonic acid (MES),2,2-Bis (hydroxymethyl)-2,2′,2″-nitrilotriethanol (bis-tris), N-(2-Acetamido) iminodiacetic acid (ADA), piperazine-N,N′-bis (2-ethanesulfonic acid) (PIPES), N-(2-Acetamido)-2-aminoethanesulfonic acid (ACES), 3-(N-morpholinyl)-2-hydroxypropanesulfonic acid sodium salt (MOPSO), 1,3-bis (tris (hydroxymethyl) methylamino) propane (bis-trispropane), N,N-Bis (2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), 2-[[1,3-dihydroxy-2-(hydroxymethyl) propan-2-yl]amino]ethanesulfonic acid (TES), 3-(Bis (2-hydroxyethyl) amino)-2-hydroxypropane-1-sulfonic acid (DIPSO), 3-[[1,3-dihydroxy-2-(hydroxymethyl) propan-2-yl]amino]-2-hydroxypropane-1-sulfonic acid (TAPSO), Trizma, piperazine-1,4-bis (2-hydroxypropanesulfonic acid) dihydrate (POPSO), 3-[4-(2-Hydroxyethyl)-1-piperazinyl]propanesulfonic acid (HEPPS), N-(2-Hydroxy-1,1-bis (hydroxymethyl) ethyl) glycine (TRICINE), glycylglycine (GLY-GLY), 2-(Bis (2-hydroxyethyl) amino) acetic acid (BICINE), N-(2-hydroxyethyl) piperazine-N′-(4-butanesulfonic acid) (HEPBS), 3-[[1,3-dihydroxy-2-(hydroxymethyl) propan-2-yl]amino]propane-1-sulfonic acid (TAPS), 2-amino-2-methyl-1,3-propanediol (AMPD), N-(1,1-Dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid (AMPSO), Ncyclohexyl-2-aminoethanesulfonic acid (CHES), N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid (CAPSO), 1-amino-2-methyl-1-propanol (AMP), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), 4-(cyclohexylamino)-1-butanesulfonic acid (CABS), Lysogeny broth, a biologically or physiologically relevant salt, or a combination thereof.
18. The method of claim 3 , wherein the target molecule is selected from the group consisting of a nucleic acid molecule, a protein, a lipid, a biological metabolite, and combinations thereof.