Granted Patent B2

US 12,213,730 · App. 18/105,636 · Granted Feb 4, 2025

Inventors: Vladimir Pavlovich Zharov (Little Rock, AR); Ekaterina Galanzha (Little Rock, AR)

Assignee: BioVentures, LLC

A61B18/245A61B5/0059A61B5/0095A61B5/02007A61B5/14535A61B5/1495A61B5/412A61B5/415A61B5/416A61B5/418A61B5/4839A61B5/7282A61B5/7405A61B5/742A61B5/7455A61B5/746A61B8/06A61B8/481A61K49/22B03C1/288B03C1/30G01N21/1702G01N29/2418G01N29/4427A61B8/08A61B2018/0041A61B2018/00577A61B2018/0088A61B18/1815A61B18/20B03C2201/06B03C2201/26G01N15/1425G01N15/1434G01N15/147G01N2015/1477G01N2201/0221

Abstract

Claims (55)

1. A method for continuous monitoring of a circulatory vessel of a living organism, the method comprising:

pulsing circulating target objects comprising red blood cells and at least one clot within the circulatory vessel with at least one pulse of laser energy at a first pulse wavelength ranging between 400 nm and 2500 nm, wherein the first pulse wavelength induces a photoacoustic signal from the circulating target objects, wherein the at least one clot is a hemoglobin-rich red clot, a platelet-rich white clot, or a combination thereof and wherein the hemoglobin-rich clot is light absorbing and the platelet-rich white clot is non-absorbing;

obtaining a photoacoustic pattern induced by the at least one pulse of laser energy, wherein the photoacoustic pattern comprises at least one photoacoustic signal comprising a blood background signal produced by the red blood cells;

simultaneously obtaining fluorescence, scattering, and photothermal signals from the target objects, via a photodetector;

processing photoacoustic, fluorescence, scattering, and photothermal signals;

analyzing a combination of the photoacoustic pattern and the fluorescence, scattering, and/or photothermal signals to determine the presence of the at least one clot, wherein analyzing the photoacoustic pattern comprises determining the presence of positive and negative contrast peaks in the photoacoustic pattern indicating the presence of a red clot, a white clot, or a combination thereof, wherein the negative contrast in the photoacoustic pattern below the blood background signal indicates the presence of the white clot, the positive contrast in the photoacoustic pattern above the blood background signal indicates the presence of the red clot, and a combination of positive contrast and negative contrast indicates the presence of a combination of red and white clots; and

producing a detection signal when the photoacoustic pattern and the fluorescence signal, scattering signal, photothermal signal, or combinations thereof indicates the presence of a clot.

2. The method of claim 1 , wherein processing photoacoustic, fluorescence, scattering, and photothermal signals comprises signal filtering and signal averaging to eliminate signal fluctuations due to breathing, blood pulse, or other body movements or signal noise using a high-pass filter, a low-pass filter, and combinations thereof.

3. The method of claim 1 , wherein analyzing the combination of the photoacoustic pattern and the fluorescence includes analyzing at two wavelengths photoacoustic signal shapes, widths, and rates of signal detection to distinguish a composition, size, and concentration of clots.

4. The method of claim 1 , wherein the circulatory vessel includes one of capillaries, arterioles, venules, arteries, veins including a carotid artery and lymphatic vessels with diameters ranged between about 10 μm and about 2 cm.

5. The method of claim 4 , wherein the circulatory vessel is located in various organs and tissues, including, but not limited to one or more of skin, lips, eyelid, interdigital membrane, retina, ear, nail pad, scrotum, brain, breast, prostate, lung, colon, spleen, liver, kidney, pancreas, heart, testicles, ovaries, lungs, uterus, skeletal muscle, smooth muscle, and bladder.

6. The method of claim 1 , the method further comprising:

simultaneously irradiating circulatory vessels at different depths;

measuring photoacoustic signals from blood vessels at different depths by determining a signal delay; and

determining the depths of the blood vessels based on the signal delay.

7. The method of claim 1 , the method further comprising:

determining the presence and extent of the at least one clot;

diagnosing a stroke, heart attack, or other clot-related disorders based on the presence and extent of the at least one clot; and

directing an application of an anti-clot treatment.

8. The method of claim 1 , the method further comprising issuing an alarm after receiving the detection signal indicating an alarm condition; wherein the alarm condition is the detection signal of sufficient magnitude or two or more detection signals at sufficiently high frequency, and wherein the alarm is a continuous or intermittent visual display, audible sound, vibration, or signal to summon medical assistance.

9. The method of claim 7 , the method further comprising:

sending a communication to a hospital, medical center, or medical assistance personal to summon medical assistance, wherein the communication is sent by wireless signals, cellular signals, or wireless Internet signals; and

administering an anti-clotting treatment based on a processed signal quantity of the detection signal, wherein the processed signal quantity is signal magnitude, frequency of signals, or elapsed time since a previous detection signal, and wherein the anti-clotting treatment is an anti-clotting medication administered continuously or in discrete doses or pulsing of the clot with a high-intensity laser pulse at a laser fluence sufficient to eliminate the clot.

10. The method of claim 1 , wherein the target objects are unlabeled biological cells, platelets, red blood cells, live, apoptotic and necrotic white blood cells, unbound contrast agents, biological cells labeled using contrast agents, aggregations of cells, sickle cells, infected cells, inflamed cells, stem cells, dendritic cells, metastatic cancer cells resulting from melanoma, leukemia, breast cancer, prostate cancer, ovarian cancer, and testicular cancer, bacteria, viruses, fungal cells, protozoa, microorganisms, pathogens, animal cells, plant cells, and leukocytes activated by various antigens during an inflammatory reaction, heterogeneous clots comprising platelets, products resulting from cell metabolism or apoptosis, cytokines or chemokines associated with the response of immune system cells to infection, exotoxins and endotoxins produced during infections, specific gene markers of cells such as tyrosinase mRNA and p97 associated with cancer cells, MelanA/Mart1 produced by melanoma cells, PSA produced by prostate cancer, and cytokeratins produced by breast carcinoma, and any combination thereof.

11. The method of claim 1 , wherein the detection of the circulating clot is verified by comparison of high-speed optical imaging taken concurrently with the photoacoustic pattern.

12. The method of claim 1 , further comprising reducing background photoacoustic signals by:

delivering oxygen or nitrogen to reduce changes in blood oxygenation;

injecting standard saline solution to decrease hematocrit; and

injecting hypertonic NaCl solution to reduce changes in blood osmolarity.

13. The method of claim 1 , further comprising enriching circulating metastatic cells and clots by mechanically squeezing micro blood and lymph vessels.

14. The method of claim 10 , wherein the contrast agents are chosen from the group including indocyanine green dye, melanin, fluoroscein isothiocyanate (FITC) dye, Evans blue dye, Lymphazurin dye, trypan blue dye, methylene blue dye, propidium iodide, Annexin, Oregon Green, C3, Cy5, Cy7, Neutral Red dye, phenol red dye, AlexaFluor dye, Texas red dye, gold nanospheres, gold nanoshells, gold nanorods, gold cages, carbon nanoparticles, prefluorocarbon particles, carbon nanotubes, carbon nanohorns, magnetic nanoparticles, quantum dots, binary gold-carbon nanotube nanoparticles, multilayer nanoparticles, clustered nanoparticles, liposomes, liposomes loaded with contrast dyes, liposomes loaded with nanoparticles, micelles, micelles loaded with contrast dyes, micelles loaded with nanoparticles, microbubbles, microbubbles loaded with contrast dyes, microbubbles loaded with nanoparticles, dendrimers, aquasomes, lipopolyplexes, nanoemulsions, polymeric nanoparticles, and combinations thereof.

15. The method of claim 10 , wherein the contrast agents are conjugates with antibodies and other ligands to target objects including CD45 as a marker associated with leukocytes and CD44 as a marker of cancer stem cells.

16. The method of claim 1 , the method further comprising:

incubating at least one microbubble with an average diameter of 2-4 μm with PEG-coated gold nanoshells (GNSs);

attaching the at least one microbubble to a blood clot or activated white blood cell; and

increasing the laser energy of the laser pulses illuminating the at least one microbubble to increase emitted photoacoustic signals.

17. A device for continuous monitoring of a circulatory vessel of a living organism, comprising:

an in vivo flow cytometer, wherein the in vivo flow cytometer comprises:

a pulsed laser configured to pulse target objects comprising light absorbing at least one hemoglobin rich red clot and/or at least one non-absorbing platelet-rich white clot within the circulatory vessel with a plurality of laser pulses at a pulse wavelength ranging between about 400 nm and about 2500 nm;

an ultrasound transducer configured to receive photoacoustic signals emitted by the target objects in response to the laser pulses and generate an output;

a photodetector configured to receive fluorescence, scattering, and/or photothermal signals emitted by the target objects; and

a processor comprising an amplifier, a data recording system, and a data analysis system having data analysis software, the processor being configured to:

receive the output from the ultrasound transducer and the fluorescence, scattering, and/or photothermal signals from the photodetector, wherein the output comprises a photoacoustic pattern;

analyze a combination of the photoacoustic pattern, the fluorescence signal, the scattering signal, and/or the photothermal signal to determine the presence of the at least one red clot, wherein analyzing the photoacoustic pattern comprises determining the presence of positive and negative contrast peaks in the photoacoustic pattern, wherein the negative contrast in the photoacoustic pattern below a background photoacoustic signal indicates the presence of the white clot, the positive contrast in the photoacoustic pattern above the background photoacoustic signal indicates the presence of the red clot, and a combination of positive contrast and negative contrast indicates the presence of a combination of red and white clots; and

produce a detection signal when the photoacoustic pattern and the fluorescence signal, scattering signal, photothermal signal, or combinations thereof indicates the presence of a clot.

18. A self-contained wearable device for continuous monitoring of a circulatory vessel of a living organism, comprising:

an in vivo flow cytometer;

a clot monitoring system configured to:

emit a pulsed laser at target objects within the circulatory vessel;

receive one or more signals comprising a photoacoustic pattern;

analyze the photoacoustic pattern to determine a presence of at least one clot within the circulatory vessel based on a presence of positive and negative contrast peaks in the photoacoustic pattern wherein the negative contrast in the photoacoustic pattern below a blood background signal indicates the presence of a white clot, the positive contrast in the photoacoustic pattern above the blood background signal indicates the presence of a red clot, and a combination of positive contrast and negative contrast indicates the presence of a combination of red and white clots; and;

produce a detection signal when the photoacoustic pattern indicates the presence of a clot;

an alarm system to provide a warning to the living organism and/or summon medical attention; and

a power source,

wherein the in vivo flow cytometer, the clot monitoring system, the alarm system, and the power source are configured to be removably attachable to an appendage of the living organism using straps, adhesive patches, or adhesive strips, and wherein the appendage is chosen from a neck, wrist, finger, forearm, upper arm, hand, foot, ankle, lip, ear, chest, abdomen, eye head, scalp, or leg.

Assignments (1)

Continuity (7)

References Cited (272)