# US 11,441,999 B2 · Assemblies and methods for reducing optical crosstalk in particle processing systems > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,441,999 B2 - Document ID: US11441999B2 - Status: Granted - Kind: B2 - Application number: 16/532,047 - Filed: 2019-08-05 - Published: 2019-11-28 - Granted: 2022-09-13 - Earliest claimed: 2013-03-14 - Assignees: Cytonome/St, LLC - Applicants: Cytonome/St, LLC - Inventors: Johnathan Charles Sharpe; Donald Francis Perrault, Jr.; Emanuel Tito Mendes Machado; Blair D. Morad - Web page: https://patentgrep.com/patent/US11441999B2 ## CPC - G01N 15/1434: Optical arrangements - G01N 15/1429: Signal processing - G01N 15/147: {the analysis being performed on a sample stream} - G01N 15/10: Investigating individual particles - G01N 15/14: Optical investigation techniques, e.g. flow cytometry - 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 The present disclosure relates to optical crosstalk reduction in particle processing (e.g., cytometry including flow cytometry using microfluidic based sorters, drop formation based sorters, and/or cell purification) systems and methods in order to improve performance. More particularly, the present disclosure relates to assemblies, systems and methods for minimizing optical crosstalk during the analyzing, sorting, and/or processing (e.g., purifying, measuring, isolating, detecting, monitoring and/or enriching) of particles (e.g., cells, microscopic particles, etc.). The exemplary systems and methods for crosstalk reduction in particle processing systems (e.g., cell purification systems) may be particularly useful in the area of cellular medicine or the like. The systems and methods may be modular and used singly or in combination to optimize cell purification based on the crosstalk environment and specific requirements of the operator and/or system. ## Claims ### Claim 1 A particle processing system comprising: a radiation source system configured to simultaneously optically interrogate a first interrogation region with light at a first modulated signal frequency and a second interrogation region with light at a second modulated signal frequency; a signal detection system including a detector configured to receive an interleaved optical signal including a first optical signal emitted from the first interrogation region and a second optical signal emitted from the second interrogation region; and an electronic system configured to separate the first optical signal from the detected interleaved optical signal based on a recognition of the first and second modulated signal frequencies. ### Claim 2 (depends on 1) The particle processing system of claim 1, wherein the electronic system is configured to mathematically deconvolve the interleaved optical signal. ### Claim 3 (depends on 1) The particle processing system of claim 1, wherein the electronic system includes a demultiplexer for separating the interleaved optical signal. ### Claim 4 (depends on 1) The particle processing system of claim 1, wherein the electronic system includes a modulation controller supplying modulation frequencies to the radiation source system and to the signal detection system. ### Claim 5 (depends on 1) The particle processing system of claim 1, wherein the first interrogation region and the second interrogation region have particles flowing therethrough, and wherein a particle flowing through the first interrogation region is illuminated with a plurality of illumination pulses at the first modulated signal frequency and a particle flowing through the second interrogation region is illuminated with a plurality of illumination pulses at the second modulated signal frequency. ### Claim 6 (depends on 1) The particle processing system of claim 1, wherein the radiation source system includes at least two lasers. ### Claim 7 (depends on 6) The particle processing system of claim 6, wherein each of the lasers is independently triggered by a modulation controller. ### Claim 8 (depends on 1) The particle processing system of claim 1, further comprising one of an acousto-optical modulator and an electro-optical modulator to modulate the signal frequency interrogating the first interrogation region. ### Claim 9 (depends on 1) The particle processing system of claim 1, wherein the first interrogation region and the second interrogation region correspond to adjacent microfluidic channels. ### Claim 10 (depends on 1) The particle processing system of claim 1, wherein the electronic system is further configured to separate the second optical signal received from the second interrogation region from the detected interleaved optical signal. ### Claim 11 (depends on 1) The particle processing system of claim 1, wherein the radiation source system interrogates the first interrogation region and the second interrogation region over a single spectral domain. ### Claim 12 (depends on 1) The particle processing system of claim 1, wherein the first interrogation region and the second interrogation region correspond to interrogation sites within a single microfluidic channel. ### Claim 13 (depends on 1) The particle processing system of claim 1, wherein the radiation source system includes an illumination beam spatial modulator to sequentially optically interrogate each of the first interrogation region and the second interrogation region with the predetermined time delay. ### Claim 14 A particle processing system, comprising: means for emitting light radiation to simultaneously optically interrogate a first interrogation region with light at a first modulated signal frequency and a second interrogation region with light at a second modulated signal frequency; signal detection means configured to receive an interleaved optical signal including a first optical signal emitted from the first interrogation region and a second optical signal emitted from the second interrogation region; and electronic means to separate the first optical signal from the detected interleaved optical signal based on a recognition of the first of the second modulated signal frequencies. ### Claim 15 (depends on 14) The particle processing system of claim 14, wherein the electronic means is configured to mathematically deconvolve the interleaved optical signal. ### Claim 16 (depends on 14) The particle processing system of claim 14, wherein the electronic means uses demultiplexing to separate the interleaved optical signal. ### Claim 17 (depends on 14) The particle processing system of claim 14, wherein the electronic means supplies modulation frequencies to the means for emitting light radiation and the signal detection means. ### Claim 18 (depends on 14) The particle processing system of claim 14, wherein the first interrogation region and the second interrogation region have particles flowing therethrough, and wherein a particle flowing through the first interrogation region is illuminated with a plurality of illumination pulses at the first modulated signal frequency and a particle flowing through the second interrogation region is illuminated with a plurality of illumination pulses at the second modulated signal frequency. ### Claim 19 (depends on 14) The particle processing system of claim 14, wherein the means for emitting light radiation includes at least two lasers. ### Claim 20 (depends on 14) The particle processing system of claim 14, wherein the means for emitting light radiation interrogates the first interrogation region and the second interrogation region over a single spectral domain.