Use of focused light scattering techniques in biological applicationa
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
G01N-015/02
G01N-021/03
G01N-015/06
G01N-015/14
G01N-015/00
출원번호
US-0502941
(2009-07-14)
등록번호
US-8828737
(2014-09-09)
발명자
/ 주소
Gabriel, Don
출원인 / 주소
Invitrox, Inc.
대리인 / 주소
Hulquist, PLLC
인용정보
피인용 횟수 :
2인용 특허 :
9
초록▼
Methods for using focused light scattering techniques for the optical sensing of biological particles suspended in a liquid medium are disclosed. The optical sensing enables one to characterize particles size and/or distribution in a given sample. This, in turn, allows one to identify the biological
Methods for using focused light scattering techniques for the optical sensing of biological particles suspended in a liquid medium are disclosed. The optical sensing enables one to characterize particles size and/or distribution in a given sample. This, in turn, allows one to identify the biological particles, determine their relative particle density, detect particle shedding, and identify particle aggregation. The methods are also useful in screening and optimizing drug candidates, evaluating the efficacy and dosage levels of such drugs, and in personalized medicine applications.
대표청구항▼
1. A method of identifying a biological particle of interest in a sample medium, said method comprising: a) passing a sample medium, which may or may not include a biological particle of interest, past a focused light beam formed by passing a collimated light beam through an acceptance aperture to n
1. A method of identifying a biological particle of interest in a sample medium, said method comprising: a) passing a sample medium, which may or may not include a biological particle of interest, past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,b) using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, andc) identifying the presence or absence of the biological particle of interest with a known size, by comparing the sizes of the particles in the sample medium with the known size of the particle of interest,wherein if particles in the sample medium are identified as having a size in the range of the size of known particles of interest, then the sample medium is said to include said known particles of interest, andwherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, andwhen the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both. 2. The method of claim 1, wherein, after an initial determination is made that a particle of interest is present in the sample medium, a confirmatory assay is performed. 3. The method of claim 2, wherein the confirmatory assay involves taking an Electrophoretic Quasi-Elastic Light Scattering (EQELS) spectra of the sample medium, and verifying the existence of a biological particle by identifying a feature, wherein the feature is known to be associated with the biological particle, and the presence of which confirms the identity of the biological particle. 4. The method of claim 2, wherein the confirmatory assay comprises incubating the sample medium with a compound known to bind to the biological particle of interest, wherein the compound is covalently linked to a microparticle or nanoparticle. 5. The method of claim 4, wherein the confirmatory assay comprises detecting a conjugate of the biological particle of interest with the compound bound to the microparticle or nanoparticle. 6. The method of claim 2, wherein the initial determination identifies an initial concentration of the particle of interest, and the confirmatory assay comprises: a) treating the sample medium with a compound known to kill a type of cell, wherein the biological particle of interest is the type of cell that is killed by the compound, andb) performing a focused light scattering analysis on the treated sample medium to determine whether the concentration of the particle of interest is lowered. 7. The method of claim 1, wherein the biological particle has a size ranging from about 0.1 μm to about 20 μm. 8. The method of claim 1, wherein the sample medium comprises one or more fluids selected from the group consisting of blood, blood products, water, cerebrospinal fluid, ascites, pleural fluid, and synovial fluid. 9. The method of claim 1, wherein the width of the focused light beam is substantially one half the width of the largest particle to be sized. 10. A method of identifying a biological particle of interest in a sample medium, said method comprising: a) passing a sample medium, which may or may not include a biological particle of interest, past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,b) using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, andc) identifying the presence or absence of the biological particle of interest with a known size, by comparing the sizes of the particles in the sample medium with the known size of the particle of interest,wherein if particles in the sample medium are identified as having a size in the range of the size of known particles of interest, then the sample medium is said to include said known particles of interest, andwherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, andwhen the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both,wherein the biological particle of interest is a lymphocyte, erythrocyte, B-cell, T-cell, neutrophil, monocyte, bacteria, fungi, virus, or protozoa. 11. A method of identifying a biological particle of interest in a sample medium, said method comprising: a) passing a sample medium, which may or may not include a biological particle of interest, past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,b) using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, andc) identifying the presence or absence of the biological particle of interest with a known size, by comparing the sizes of the particles in the sample medium with the known size of the particle of interest,wherein if particles in the sample medium are identified as having a size in the range of the size of known particles of interest, then the sample medium is said to include said known particles of interest, andwherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, andwhen the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both,wherein the biological particle is selected from the group consisting of tumor cells, red blood cells, white blood cells, granulocytes, platelets, monocytes, neutrophils, lymphocytes, cancer cells, stem cells, bacteria, viruses, and fungi. 12. A method of identifying a biological particle of interest in a sample medium, said method comprising: a) passing a sample medium, which may or may not include a biological particle of interest, past a focused light beam, wherein the width of the focused light beam is configured to be narrowed to substantially one half the width of the largest particle to be sized,b) using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, andc) identifying the presence or absence of the biological particle of interest with a known size, by comparing the sizes of the particles in the sample medium with the known size of the particle of interest,wherein if particles in the sample medium are identified as having a size in the range of the size of known particles of interest, then the sample medium is said to include said known particles of interest, andwherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, andwhen the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both.
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