# US 2025/0147006 A1 · Multi-depth spiral milli fluidic device for whole mount zebrafish antibody staining > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0147006 A1 - Document ID: US20250147006A1 - Status: Published application - Kind: A1 - Application number: 18/939,842 - Filed: 2024-11-07 - Published: 2025-05-08 - Earliest claimed: 2023-11-07 - Assignees: The Regents of the University of California - Applicants: The Regents of the University of California - Inventors: Songtao Ye; Chih-Wen Ni - Web page: https://patentgrep.com/patent/US20250147006A1 ## CPC - G01N 33/5014: {for testing toxicity} - C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)} - C12M 29/06: {Nozzles; Sprayers; Spargers; Diffusers (per se B01F23/231, B01J19/26)} - C12M 41/46: {of cellular or enzymatic activity or functionality, e.g. cell viability} - G01N 33/5082: {Supracellular entities, e.g. tissue, organisms} - 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) - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} ## Abstract Fluidic systems, devices and methods are provided for separating and sequestering particles from a fluid flow within traps in a fluidic chip is provided. The fluidic platform is particularly suited for parallel live zebrafish embryo studies providing automated zebrafish embryo trapping and flow through culture as well as whole mount zebrafish antibody staining functions. The zebrafish on a chip testing platform uses a chaotic hydrodynamic trapping process to trap and retain zebrafish embryos in a consistent body orientation (i.e., head pointed inward) without any external adjustments. The system and apparatus can also be adapted to be a multifunctional concentration gradient generator (CGG) that can be used to automatically immobilize dechorionated zebrafish embryos and generate chemical gradients for acute fish embryo toxicity (FET) tests. ## Claims ### Claim 1 A fluidic device apparatus, comprising: (a) a fluidic chip body with a central fluid collection reservoir defined by an inner wall of an arcuate circumferential channel, said channel open at one end to the central fluid collection reservoir with an opening, said inner wall of said arcuate channel having a plurality of particle traps; (b) at least one fluid inlet fluidly coupled to said arcuate channel; and (c) a fluid outlet fluidly coupled to said central fluid collection reservoir; (d) wherein individual particles in a flow of fluid through the arcuate annular channel are trapped in the particle traps. ### Claim 2 (depends on 1) The apparatus of claim 1, wherein said arcuate channel forms a single loop spiral, said opening of the channel to the central fluid collection reservoir at a distal end of the arcuate channel. ### Claim 3 (depends on 1) The apparatus of claim 1, wherein said opening of said arcuate channel has a diameter that is smaller than a diameter of the arcuate channel. ### Claim 4 (depends on 3) The apparatus of claim 3, further comprising a flow controller disposed within said arcuate channel configured to reduce fluid flow from the arcuate channel into the central fluid collection reservoir. ### Claim 5 (depends on 1) The apparatus of claim 1, wherein said particle traps of said inner wall of said arcuate channel have a trap body open to the channel and a narrower trap nozzle that is open to the central fluid collection reservoir, said trap configured to allow fluid to flow from the arcuate channel through the trap body and trap nozzle to the central fluid collection reservoir. ### Claim 6 (depends on 5) The apparatus of claim 5, wherein said trap body open to the arcuate channel has walls that are perpendicular to a surface of the inner wall of the arcuate channel. ### Claim 7 (depends on 5) The apparatus of claim 5, wherein said trap body open to the arcuate channel of has an opening with angular edges. ### Claim 8 (depends on 5) The apparatus of claim 5, wherein said trap body open to the arcuate channel of has an opening with an angular edge and a rounded edge, said rounded edge facing a direction of fluid flow towards a distal end of the arcuate channel. ### Claim 9 (depends on 1) The apparatus of claim 1, wherein said at least one fluid inlet comprises: a first fluidic input fluidly coupled to a proximal end of the arcuate channel; a second fluidic input fluidly coupled to a proximal end of the arcuate channel; and a carrier fluid input fluidly coupled to a proximal end of the arcuate channel. ### Claim 10 A system for separating and retaining particles, the system comprising: (a) a fluidic chip with an arcuate channel with an outer wall and an inner wall, said arcuate channel forming a single loop spiral, said inner wall of said channel defining a central fluid collection reservoir, said arcuate channel open at one end to the central fluid collection reservoir, said inner wall of said arcuate channel having a plurality of particle traps; at least one fluid inlet fluidly coupled to said arcuate channel; and a fluid outlet fluidly coupled to said central fluid collection reservoir; (b) a pump; (c) a fluidic circuit between the pump and the fluid outlet of the central fluid collection reservoir of the fluidic chip; and (d) a controller configured to control actuation of the pump. ### Claim 11 (depends on 10) The system of claim 10, wherein said pump comprises a peristaltic pump and a pulse dampener. ### Claim 12 (depends on 10) The system of claim 10, further comprising: a closed fluidic circuit between the fluid outlet and the fluid inlet of the fluidic chip; and wherein fluid flows through the fluid inlet, the arcuate channel, the fluid collection reservoir and the fluid outlet can be recycled. ### Claim 13 (depends on 10) The system of claim 10: wherein said controller is configured to control a pressure of fluid entering the fluid inlet and a pressure of the fluid exiting the fluid outlet within the collection reservoir; and wherein said controller can create a pressure differential between the arcuate channel and the central fluid collection reservoir. ### Claim 14 (depends on 10) The system of claim 10, said pump further comprising: a fluid heating element; a temperature sensor; and a fluid temperature control circuit. ### Claim 15 (depends on 10) The system of claim 10, further comprising: one or more syringe pumps fluidly coupled to said at least one fluid inlet of said fluidic chip; a fluidic circuit with valves fluidly coupled to and input and an output side of the pump that can switch between a close-loop pumping configuration and an open-loop pumping configuration, said input side valves of said fluidic circuit also coupled to a first waste tank and said output side valves coupled to a second waste tank; wherein open-loop pumping fluid flow through the fluid inlet, the arcuate channel, the fluid collection reservoir and the fluid outlet is directed to a waste tank; and wherein close-loop pumping fluid flow through the fluid inlet, the arcuate channel, the fluid collection reservoir and the fluid outlet can be recycled through the fluid inlet. ### Claim 16 A method for particle separations in a fluid flow, the method comprising: (a) providing a fluidic chip with an arcuate annular channel defined by an outer wall and an inner wall, said channel open at one end to a central fluid collection reservoir, said inner wall of said channel having a plurality of particle traps, at least one fluid inlet fluidly coupled to said arcuate channel; and a fluid outlet inlet fluidly coupled to said central fluid collection reservoir; (b) flowing a fluid with particles for separation through said fluid inlet and arcuate channel; and (c) trapping individual particles within said particle traps from the fluid flow by controlling the flow of fluid into the channel and out of the collection reservoir. ### Claim 17 (depends on 16) The method of claim 16, further comprising: dispensing a first fluid through a first fluid inlet to said arcuate channel; dispensing a second fluid through a second fluid inlet to said arcuate channel; mixing said first and second fluids with a carrier fluid in the arcuate channel to produce mixed fluids; and flowing the mixed fluids through the arcuate channel, traps and central fluid collection reservoir; wherein a gradient of mixed fluids is generated in said arcuate channel. ### Claim 18 (depends on 17) The method of claim 17, further comprising: flushing said arcuate channel, traps and central fluid collection reservoir with carrier fluid after flowing said mixed fluids through the chip. ### Claim 19 (depends on 16) The method of claim 16, further comprising controlling fluid temperature of the fluid entering the fluid inlet of the chip. ### Claim 20 (depends on 16) The method of claim 16, further comprising recycling particle containing fluid exiting the fluid outlet back to the fluid inlet of the fluidic chip.