# US 2014/0152986 A1 · Methods and apparatus for determining characteristics of particles > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0152986 A1 - Document ID: US20140152986A1 - Status: Published application - Kind: A1 - Application number: 14/157,961 - Filed: 2014-01-17 - Published: 2014-06-05 - Earliest claimed: 2006-08-30 - Inventors: Michael Trainer - Web page: https://patentgrep.com/patent/US20140152986A1 ## CPC - G01N 15/0211: {Investigating a scatter or diffraction pattern} - G01N 15/0205: by optical means - G01N 15/1459: {the analysis being performed on a sample stream} - G01N 21/0303: {Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment (G01N21/031 - G01N21/15 take precedence)} - G01N 2015/1497: {Particle shape} - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Method and apparatus are described for improving measurements of scattered light from particles by controlling multiple scattering and coincidence count levels. The scatter path in the particle dispersion and particle concentration are adjusted to reduce multiple scattering in ensemble particle scattering measurement. And the particle dispersion volume and particle concentration are adjusted to reduce coincidence counts in single particle scattering measurements. Alignment of the optical system, for measuring scattered light, is maintained by a reflection apparatus. ## Claims ### Claim 1 Apparatus for optimizing a path length of light in a particle dispersion to improve the measurement of light scattered from particles, comprising: a) an optical system for illuminating a particle dispersion, b) a detection system comprising at least one detector for quantifying characteristics of light related to light scattered by at least one of said particles, c) a sample cell, comprising two optical windows, which confine said particle dispersion and pass light, from a light source, through said particle dispersion to illuminate at least one of said particles, d) means for adjusting a separation between said optical windows to change a path length of light in said sample cell, wherein said adjusting is performed by utilizing an electronic controlled actuator, and e) means for controlling said actuator based upon characteristics of light scattered from at least one of said particles. ### Claim 2 Apparatus for adjusting the concentration of particles, comprising: a) means for introducing a plurality of particles into a first flow loop, the particles being contained in a fluid, and means for circulating the fluid through the first flow loop, and b) valve means for selectively allowing some of the fluid in the first flow loop to enter a second flow loop, distinct from the first flow loop, the second flow loop including means for counting and/or analyzing at least one particle flowing in the second flow loop, wherein the valve means comprises means for transferring fluid from said first flow loop to fluid in said second flow loop, and wherein the valve means transfers an amount of fluid which produces an optimum count rate of particles of interest and/or an optimum particle concentration, measured from particles in said second flow loop. ### Claim 3 Apparatus for maintaining alignment in an optical system for measuring scattered light comprising: a) illumination means for illuminating at least one particle in a particle sample region, b) a detection means comprising at least one detector for quantifying characteristics of light related to light scattered by at least one particle, and c) a particle sample region wherein said illumination means illuminates at least one particle, d) a retro-reflector means to reflect light to said detection means, e) a support means for attaching said illumination means and detection means to a common support structure, wherein the optical axis of said illumination means and the optical axis of said detection means are aligned to utilize the optical characteristics of said retro-reflector means to maintain proper alignment of light from said illumination means and/or scattered light onto at least one detector in said detection means. ### Claim 4 (depends on 1) The apparatus of claim 1, further comprising a flexible conduit attached to at least one of said windows to confine said particle dispersion while allowing motion of at least one of said windows. ### Claim 5 (depends on 1) The apparatus of claim 1, further comprising a flexible diaphragm attached to at least one of said windows to confine said particle dispersion while allowing motion of at least one of said windows. ### Claim 6 (depends on 1) The apparatus of claim 1, further comprising an apparatus for adjusting the concentration of particles, comprising: e) means for introducing a plurality of particles into a first flow loop, the particles being contained in a fluid, and means for circulating the fluid through the first flow loop, and f) valve means for selectively allowing some of the fluid in the first flow loop to enter a second flow loop, distinct from the first flow loop, the second flow loop including means for counting and/or analyzing particles flowing in the second flow loop, wherein the valve means comprises means for transferring fluid from said first flow loop to fluid in said second flow loop, and wherein the valve means transfers an amount of fluid which produces an optimum count rate of particles of interest and/or an optimum particle concentration, measured from particles in said second flow loop. ### Claim 7 (depends on 1) The apparatus of claim 1, further comprising: f) illumination means for illuminating at least one particle in a particle dispersion, g) a detection means comprising at least one detector for quantifying characteristics of light related to light scattered by at least one particle, and h) a particle sample region wherein said illumination means illuminates at least one particle, i) a retro-reflector means to reflect light to said detection means, j) a support means for attaching said illumination means and detection means to a common support structure, wherein the optical axis of said illumination means and the optical axis of said detection means are aligned to utilize the optical characteristics of said retro-reflector means to maintain proper alignment of light from said illumination means and/or scattered light onto at least one detector in said detection means. ### Claim 8 (depends on 1) The apparatus of claim 1 further comprising: f) attachment means for attaching at least one of said optical windows to said sample cell, wherein said attachment means provides for detachment and reattachment of said window from said sample cell, g) means for placing a particle dispersion sample upon at least one of said windows, while at least one of said windows is detached from said sample cell, and h) means for reattaching at least one detached window to said sample cell. ### Claim 9 (depends on 1) The apparatus of claim 1, the optical windows including optical paths for allowing light to enter the sample cell and to leave the sample cell after interacting with particles, the sample cell also defining a volume within which particles pass through the sample cell, wherein the sample cell includes at least one optically transmitting shaped object, the shaped object being positioned outside of a desired portion of said volume so as to prevent particles from passing through a volume occupied by said object and wherein said occupied volume comprises the portion of illuminated volume in the particle dispersion outside of the desired portion of said volume, and wherein said object is shaped such that particles, which are too large to pass through the desired portion of said volume, pass around said object in the particle dispersion flow, and wherein said object has acceptable optical transmission at the optical wavelength of said scattered light. ### Claim 10 (depends on 3) The apparatus of claim 3, further comprising: f) a sample cell, comprising two optical windows, which confine said particle dispersion and pass light, from said illumination means, through said particle dispersion to illuminate at least one of said particles. ### Claim 11 (depends on 3) The apparatus of claim 3, further comprising: f) an illumination-detection module comprising said support means, said illumination means, and said detection means, and wherein said illumination-detection module is separate from said retro-reflector means, and wherein said particle sample region resides between said illumination-detection module and said retro-reflector means, and wherein the spacing between said retro-reflector and said illumination-detection module is adjusted to accommodate the size of the particle sample region. ### Claim 12 (depends on 3) The apparatus of claim 3, further comprising: f) an illumination-detection module comprising said support means, said illumination means, and said detection means, and wherein said illumination-detection module is separate from said retro-reflector means, g) a first module comprising said illumination-detection module and a first optical window, wherein said first optical window is attached to said illumination-detection module, and h) a second module comprising said retro-reflector means and a second optical window, wherein said second optical window is attached to said retro-reflector means, and wherein the particle sample region resides between said first optical window and said second optical window and wherein the distance between said first module and said second module determine a light pathlength in said particle dispersion. ### Claim 13 A method for optimizing a path length of light in a particle dispersion to improve the measurement of light scattered from at least one particle, comprising: a) illuminating a particle dispersion, b) detecting characteristics of light related to light scattered by at least one particle, c) confining said particle dispersion between two optical windows in a sample cell, d) passing light, from a light source, through said particle dispersion to illuminate at least one particle, e) adjusting a separation between said optical windows to change a path length of light in said dispersion, wherein said adjusting is performed by utilizing an electronic controlled actuator, and wherein said adjusting is based upon characteristics of light scattered from at least one particle. ### Claim 14 (depends on 13) The method of claim 13 wherein said adjusting is determined to reduce coincidence counts of particles of interest and/or to obtain a desired particle count rate of particles of interest in a particle counting scatter measurement, further comprising: a) determining a desired value of average number of particles of interest per interaction volume for said particle dispersion to provide acceptable coincidence counts, b) adjusting said separation between said optical windows to a first separation value, c) measuring a first particle dispersion average number of particles of interest per interaction volume value at said first separation value, d) calculating a desired separation between said optical windows to obtain said desired value of average number of particles of interest per interaction volume utilizing information comprising said desired value of average number of particles of interest per interaction volume, said first separation value, and said first particle dispersion average number of particles of interest per interaction volume value, e) adjusting said separation between said optical windows to the desired separation value, and f) measuring scattered light from particle dispersion within said desired separation between said optical windows and using said measured scattered light to determine information about at least one particle. ### Claim 15 (depends on 13) The method of claim 13 wherein said adjusting is determined by determining a difference in shape between functions, wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 16 (depends on 13) The method of claim 13 further comprising the steps of: a) adjusting said separation between said optical windows to provide desired coincidence counts of particles of interest, and a) scanning the particle dispersion between said optical windows by illuminating and detecting characteristics of light related to scattered light at a plurality of regions in the particle dispersion between said optical windows. ### Claim 17 (depends on 16) The method of claim 16 adapted to measure light scattering from particles with significant settling velocity, further comprising the steps of: a) adjusting said separation between said optical windows while the windows are generally parallel to gravitational force, and b) scanning the particle dispersion between said optical windows while the windows are generally perpendicular to gravitational force. ### Claim 18 (depends on 13) The method of claim 13 adapted to provide control of coincidence counts, further comprising the steps of: a) adjusting said separation between said optical windows to a first separation which allows flowing of particle dispersion between said optical windows, b) determining a desired particle concentration, at said first separation, to provide desired coincidence counts for a desired second separation between said optical windows, c) determining a desired scattering characteristic which is expected from said desired particle concentration at said first separation, d) adjusting the particle concentration during flowing of particle dispersion through a volume between said optical windows at said first separation, e) monitoring at least one characteristic of scattered light of the particle dispersion between said optical windows while adjusting said particle concentration to obtain said desired scattering characteristic, f) adjusting said separation between said optical windows to said desired second separation, and g) measuring scattered light from at least one particle in said particle dispersion between said optical windows at said desired second separation. ### Claim 19 (depends on 13) The method of claim 13 further comprising: a) determining a desired value of transmission for said particle dispersion to provide acceptable multiple scattering, b) adjusting said separation between said optical windows to a first separation value, c) measuring a first particle dispersion transmission value at said first separation value, d) calculating the desired separation between said optical windows to obtain said desired value of transmission utilizing information comprising said desired value of transmission, first particle dispersion transmission value, and said first separation value, e) adjusting said separation between said optical windows to the desired separation value, and f) measuring scattered light from at least one particle within said desired separation between said optical windows. ### Claim 20 (depends on 13) The method of claim 13, further comprising: a) adjusting a parameter to a first value, b) determining a first function with parameter set at said first value, c) adjusting said parameter to a second value, which is less than said first value, d) determining a second function with parameter set at said second value, e) determining a difference in shape between said first function and said second function, f) comparing said shape difference to a specified limit, g) if said shape difference is greater than said specified limit, repeating steps (a) through (h), while setting first function, in step (b), to said second function and while setting first value of parameter, in step (a), to said second value of parameter, and h) if said shape difference is less than or equal to said specified limit, using the function corresponding to the lowest value of the two parameter values, which produced said shape difference, to determine particle characteristics, wherein said parameter comprises a member of the group consisting of separation between said optical windows, particle concentration, and a product of particle concentration and separation between said optical windows, and wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 21 (depends on 13) The method of claim 13 further comprising the steps of: a) adjusting said separation between said optical windows to allow flow of particle dispersion between said optical windows, b) flowing particle dispersion between said optical windows, c) adjusting particle concentration to an acceptable value, if required, d) stopping flow of particle dispersion, e) adjusting said separation between said optical windows to provide low coincidence counts of particles of interest, f) scanning the particle dispersion between said optical windows by illuminating and detecting characteristics of light related to scattered light at a plurality of regions in the particle dispersion between said optical windows, and f) performing steps (a), (b), (d), (e) and (f) a plurality of times, combining the measurements from all performances to produce information from a sufficient number of particles. ### Claim 22 A method for adjusting the concentration of particles, comprising: a) introducing a plurality of particles into a first flow loop, the particles being contained in a fluid, and circulating the fluid through the first flow loop, b) utilizing a valve to selectively allow some of the fluid in the first flow loop to enter a second flow loop, distinct from the first flow loop, the second flow loop including and counting and/or analyzing at least one particle flowing in the second flow loop, wherein the valve transfers fluid from said first flow loop to fluid in said second flow loop, and wherein the valve transfers an amount of fluid which produces an optimum count rate of particles of interest and/or an optimum particle concentration, measured from particles in said second flow loop. ### Claim 23 (depends on 22) The method of claim 22 further comprising: a) determining an optimum particle dispersion transmission, b) first transferring a first volume of particle dispersion into said second flow loop from said first flow loop, c) determining a first transmission of said particle dispersion in a sample cell of said second flow loop after said first transferring, d) determining a second volume of particle dispersion for a second transferring into said second flow loop to produce said optimum particle dispersion transmission, wherein said determining utilizes information comprising values of said optimum particle dispersion transmission, said first volume of particle dispersion, and said first transmission of said particle dispersion, e) second transferring said second volume of particle dispersion into said second flow loop from said first flow loop, and f) measuring scattered light from at least one particle in said second flow loop. ### Claim 24 (depends on 22) The method of claim 22 further comprising: a) determining an optimum average number of particles of interest per interaction volume, b) first transferring a first volume of particle dispersion into said second flow loop from said first loop, c) determining a first average number of particles of interest per interaction volume in said second flow loop after said first transferring, d) determining a second volume of particle dispersion for a second transferring into said second flow loop to produce said optimum average number of particles of interest, wherein said determining utilizes information comprising values of said optimum average number of particles of interest, said first volume of particle dispersion, and said first average number of particles of interest, e) second transferring said second volume of particle dispersion into said second flow loop from said first loop, and f) measuring scattered light from at least one particle in said second flow loop. ### Claim 25 (depends on 13) The method of claim 13 further comprising adjusting a product of particle concentration and separation between said optical windows to obtain at least one member of the group consisting of desired particle coincidence counts, desired particle count rate, desired scatter signal, and desired multiple scattering. ### Claim 26 (depends on 13) The method of claim 13, further comprising a method for determining an optimum path length of light in a particle dispersion, so as to reduce inaccuracies in particle analysis caused by multiple scattering of light from particles, the method comprising the steps of: a) determining a specified value of a path length which produces excessive multiple scattering, b) adjusting said path length to a specified value, c) measuring first function values at said specified value of path length, d) changing said path length to a second value, which is less than said specified value, e) measuring second function values at said second value of said path length, f) determining a difference between said second function values and said first function values, g) if said difference is greater than a specified limit, repeating steps (b) through (f), while setting said specified value of path length, in step (b), to said second value of said path length, h) if said difference is less than or equal to a specified limit, using said second function values to determine particle characteristics, further comprising selecting said difference between said second function values and first function values to be a difference in functional shape, and wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 27 (depends on 13) The method of claim 13, further comprising: a) adjusting a parameter to a first value, wherein said first value provides a generally lowest acceptable scatter signal and low multiple scattering, b) measuring a first function of scattered light with parameter at said first value, c) adjusting said parameter to a value which is larger than the current value, d) measuring a new function of scattered light with parameter at said larger value, e) determining a difference in shape between said first function and said new function, f) comparing said shape difference to a specified limit, g) if said shape difference is less than said specified limit, repeating steps (c) through (h), h) if said shape difference is greater than or equal to said specified limit, using the scatter function corresponding to the lowest value of the two parameter values, which produced said shape difference, to determine particle characteristics, wherein said parameter comprises a member of the group consisting of separation between said optical windows, particle concentration, and a product of particle concentration and separation between said optical windows, and wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 28 (depends on 13) The method of claim 13, further comprising: a) adjusting a parameter to a first value, b) measuring a first function with parameter at said first value, c) adjusting said parameter to a second value, which is greater than said first value, d) measuring a second function with parameter at said second value, e) determining a difference in shape between said first function and said second function, f) comparing said shape difference to a specified limit, g) if said shape difference is less than said specified limit, repeating steps (a) through (h), while setting first function, in step (b), to said second function and while setting first value of parameter, in step (a), to said second value of parameter, and h) if said shape difference is greater than or equal to said specified limit, using the function corresponding to the lowest value of the two parameter values, which produced said shape difference, to determine particle characteristics, wherein said parameter comprises a member of the group consisting of separation between said optical windows, particle concentration, and a product of particle concentration and separation between said optical windows, and wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 29 (depends on 22) The method of claim 22, further comprising, a) adjusting a parameter to a first value, b) measuring a first function with parameter at said first value, c) adjusting said parameter to a second value, which is greater than said first value, d) measuring a second function with parameter at said second value, e) determining a difference in shape between said first function and said second function, f) comparing said shape difference to a specified limit, g) if said shape difference is less than said specified limit, repeating steps (a) through (h), while setting first function, in step (b), to said second function and while setting first value of parameter, in step (a), to said second value of parameter, and h) if said shape difference is greater than or equal to said specified limit, using the function corresponding to the lowest value of the two parameter values, which produced said shape difference, to determine particle characteristics, wherein said parameter comprises particle concentration, and wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 30 (depends on 22) The method of claim 22, further comprising: a) adjusting a parameter to a first value, b) measuring a first function with parameter at said first value, c) adjusting said parameter to a second value, which is less than said first value, d) measuring a second function with parameter at said second value, e) determining a difference in shape between said first function and said second function, f) comparing said shape difference to a specified limit, g) if said shape difference is greater than said specified limit, repeating steps (a) through (h), while setting first function, in step (b), to said second function and while setting first value of parameter, in step (a), to said second value of parameter, and h) if said shape difference is less than or equal to said specified limit, using the function corresponding to the lowest value of the two parameter values, which produced said shape difference, to determine particle characteristics, wherein said parameter comprises particle concentration, and wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size. ### Claim 31 (depends on 22) The method of claim 22 wherein said transferred amount of fluid is determined by determining a difference in shape between functions, wherein said function comprises a member of the group consisting of scattered light intensity as a function of scattering angle, scattered light flux as a function of different ranges of scattering angle, and particle number, or particle volume, as a function of different ranges of particle size.