WO2024074828A1

WO2024074828A1 - Systems and methods for fluid distribution

Info

Publication number: WO2024074828A1
Application number: PCT/GB2023/052568
Authority: WO
Inventors: Cesare M. Cejas; Antonio De Grazia; Richard Baxter; Ian Johnston; Christabel TAN; Nikolay DIMOV
Original assignee: Microfluidx Ltd
Priority date: 2022-10-05
Filing date: 2023-10-04
Publication date: 2024-04-11

Abstract

The present disclosure provides systems, devices, and methods for fluid distribution. The systems and devices herein can comprise an input channel configured to receive a fluid; one or more levels of branched channels fluidically connected to said input channel, wherein said one or more levels of branched channels are below said input channel; and a plurality of outputs fluidically connected to said one or more levels of branched channels, wherein said plurality of outputs are below said one or more levels of branched channels, wherein said device is configured to allow a fluid to flow in a substantially vertical direction from said input channel, through said one or more levels of branched channels, and to said plurality of outputs, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within a standard deviation of 10% of a mean volume of said plurality of outputs.

Description

SYSTEMS AND METHODS FOR FLUID DISTRIBUTION

CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63/413,528, filed October 5, 2022, which is herein entirely incorporated by reference.

BACKGROUND

[0002] Fill and Finish (F&F) in cell therapy is the process of washing, concentrating, resuspending cells at a desired concentration and in a formulated solution, and distributing the cells across a number of containers. Cells are suspended in a solution made from several reagents suitable to protect the cells against cryo-storage before infusion into the patient. The batch of cells can be broadly distributed into n containers evenly, with n ranging from <10 to several hundreds. For autologous products, n typically ranges from 2 to 5, where some containers are destined to the become patient material (n>l if several infusions are desired) and some others are kept for quality assurance.

[0003] There are various limitations with conventional F&F systems. Processes can require extensive manual input, e.g., repetitive pipetting, which can create variability in the process, e.g., the number of cells per container. Processes can be open, meaning they have to be carried out in an A grade facility (e.g., Biosafety cabinet), which can be both cumbersome and expensive. Existing automated platforms can have limited capacity (e.g., n=3), requiring multiple steps to achieve a large number of outputs. Some platforms also require output containers with minimum volumes (e.g., bags of >150mL), which is not suited for certain applications that require smaller container volumes (vials of <10mL). Other platforms, in particular robotic platforms, fill in a sequential manner, which can create differences in cell concentration as it becomes harder to maintain a stable cell concentration in an ever drawn-down volume of stock solution. Additionally, sequential filling can be lengthy, e.g., 600 vials taking one hour to fill, generating product consistency issues, e.g., from cells in the first vial having been exposed differently to the cryopreservant to the ones in last vial filled. Thus, there is a need for improved methods and systems for evenly distributing fluids in storage containers, that is adapted to a large n, can handle small volumes, and promotes product consistency through rapid processing.

SUMMARY

[0004] The present invention discloses a microfluidics-based alternative to standard F&F systems. The systems and methods described herein can function as a plug and play system where a stock solution is injected in a primary input, and a branching design helps to equally distribute the concentration of the stock solution at each output. The systems and methods described herein can evenly divide input cells and reagents into output containers with no more than 10% standard deviation in cell count and no more than 5% standard deviation in total fill volume across all containers filled in a single batch.

[0005] In an aspect, provided herein is avertical-flow fluid distribution device, comprising: (a) an input channel configured to receive a fluid; (b) one or more levels of branched channels fluidically connected to the input channel, wherein the one or more levels of branched channels are below the input channel; and (c) a plurality of outputs fluidically connected to the one or more levels of branched channels, wherein the plurality of outputs are below the one or more levels of branched channels, wherein the device is configured to allow a fluid to flow in a substantially vertical direction from the input channel, through the one or more levels of branched channels, and to the plurality of outputs, and wherein the device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of the fluid in each output is within 10% of a mean volume of the plurality of outputs.

[0006] In some cases, each level of branched channels is separated from an upper channel by a binary split configured to distribute the fluid between two lower channels. In some cases, each binary split slopes downward from the upper channel directly above it at the same angle. In some cases, the angle is about 100° to about 140°. In some cases, the angle is about 115° to about 125°. [0007] In some cases, the device further comprises one or more vents comprising a sterile filter. In some cases, an inner surface of the input channel or the one or more levels of branched channels comprises a raised structure configured to homogenize flow of the fluid through the input channel. In some cases, the raised structure comprises a herringbone pattern. In some cases, the system comprises an optically transparent material.

[0008] In some cases, the fluid comprises solids. In some cases, a concentration of solids in each output is within 15% of a mean concentration of solids of the plurality of outputs. In some cases, a concentration of solids in each output is within 10% of a mean concentration of solids of the plurality of outputs. In some cases, the solids comprise one or more cells. In some cases, the device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of the fluid in each output is within 5% of a mean volume of the plurality of outputs.

[0009] In some cases, the number of outputs is an odd-number (e.g. 1 input to 3 outputs). In some cases, a branched channel of the plurality of branched channels has a curved surface. In some cases, the input channel is located on a center axis of the device.

[0010] In another aspect, provided herein is a vertical -flow fluid distribution device, comprising: (a) an input channel configured to receive a fluid; (b) two or more levels of branched channels fluidically connected to the input channel, wherein the two or more levels of branched channels are below the input channel, and wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute the fluid between two lower channels; and (c) a plurality of outputs fluidically connected to the two or more levels of branched channels, wherein the plurality of outputs are below the two or more levels of branched channels; wherein each binary split slopes downward from the upper channel directly above it at the same angle.

[0011] In some cases, the angle is about 100° to about 140°. In some cases, the angle is about 115° to about 125°. In some cases, the device further comprises one or more vents comprising a sterile filter. In some cases, an inner surface of the input channel or the two or more levels of branched channels comprises a raised structure configured to homogenize flow of the fluid through the input channel. In some cases, the raised structure comprises a herringbone pattern. In some cases, the system is comprised of an optically transparent material.

[0012] In some cases, the fluid comprises solids. In some cases, a concentration of solids in each output is within 15% of a mean concentration of solids of the plurality of outputs. In some cases, a concentration of solids in each output is within 10% of a mean concentration of solids of the plurality of outputs. In some cases, the solids comprise one or more cells.

[0013] In some cases, the device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of the fluid in each output is within 10% of a mean volume of the plurality of outputs. In some cases, the device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of the fluid in each output is within 5% of a mean volume of the plurality of outputs.

[0014] In another aspect, provided herein is a method for distributing a fluid, comprising: (a) providing a vertical-flow fluid distribution device comprising an input channel, one or more levels of branched channels fluidically connected to the input channel; (b) directing a fluid to the input channel, thereby allowing the fluid to flow in a substantially vertical direction from the input channel through the one or more levels of branched channels; and (c) producing a plurality of output streams, wherein the volume of the fluid in each outlet stream is within 10% of a mean volume of the plurality of outlet streams.

[0015] In some cases, each level of branched channels is separated from an upper channel by a binary split configured to distribute the fluid between two lower channels. In some cases, each binary split slopes downward from the upper channel directly above it at the same angle. In some cases, the angle is about 100° to about 140°. In some cases, the angle is about 115° to about 125°. In some cases, the method further comprises releasing air from the vertical-flow fluid distribution system through one or more vents, which consists of a sterile filter. In some cases, an inner surface of the input channel or the one or more levels of branched channels comprises a raised structure configured to homogenize flow of the fluid through the input channel. In some cases, the raised structure comprises a herringbone pattern. In some cases, the vertical-flow fluid distribution device is comprised of an optically transparent material.

[0016] In some cases, the fluid comprises solids. In some cases, a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of the plurality of outlet streams. In some cases, a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of the plurality of outlet streams. In some cases, the solids comprise one or more cells. In some cases, the method further comprises optically observing a flow pattern of the fluid through the optically transparent material.

[0017] In another aspect, provided herein is a vertical -flow fluid distribution system, comprising: (a) a primary fluid distribution device configured to receive a fluid; and (b) a plurality of secondary fluid distribution devices fluidically connected to the primary fluid distribution device, wherein the primary fluid distribution device is configured to distribute the fluid between each secondary fluid distribution device; wherein the vertical-flow distribution system is configured to output the fluid in a plurality of output streams, and wherein a volume of the fluid in each outlet stream is within 10% of a mean volume of the plurality of outlet streams.

[0018] In some cases, the primary fluid distribution device comprises: an input channel configured to receive a fluid and one or more levels of branched channels fluidically connected to the input channel, wherein the one or more levels of branched channels are below the input channel. In some cases, each level of branched channels is separated from an upper channel by a binary split configured to distribute the fluid between two lower channels. In some cases, each binary split slopes downward from the upper channel directly above it at the same angle. In some cases, the angle is about 100° to about 140°. In some cases, the angle is about 115° to about 125°. In some cases, an inner surface of the input channel or the one or more levels of branched channels comprises a raised structure configured to homogenize flow of the fluid through the input channel. In some cases, the raised structure comprises a herringbone pattern. In some cases, the primary fluid distribution device or the plurality of secondary fluid distribution devices are comprised of an optically transparent material.

[0019] In some cases, a secondary fluid distribution device of the plurality of secondary fluid distribution devices comprises an input channel configured to receive a fluid and one or more levels of branched channels fluidically connected to the input channel, wherein the one or more levels of branched channels are below the input channel. In some cases, each level of branched channels is separated from an upper channel by a binary split configured to distribute the fluid between two lower channels. In some cases, each binary split slopes downward from the upper channel directly above it at the same angle. In some cases, the angle is about 100° to about 140°. In some cases, the angle is about 115° to about 125°. In some cases, an inner surface of the input channel or the one or more levels of branched channels comprises a raised structure configured to homogenize flow of the fluid through the input channel. In some cases, the raised structure comprises a herringbone pattern. [0020] In some cases, the primary fluid distribution device is fluidically connected to the plurality of second fluid distribution devices through a plurality of tubes. In some cases, a secondary fluid distribution device of the plurality of secondary fluid distribution devices outputs an outlet streams an outlet stream of the plurality of outlet streams into a container. In some cases, when in the container, the output stream of the plurality of outlet streams is not fluidically connected with another outlet stream of the plurality of outlet streams. In some cases, the container is fluidically connected to the secondary a secondary fluid distribution device of the plurality of secondary fluid distribution devices via tubing. In some cases, the container comprises a centrifuge tube, cryo-bag, freezing bag, freeze-drying vial, or injection vial.

[0021] In some cases, the fluid comprises solids. In some cases, a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of the plurality of outlet streams. In some cases, a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of the plurality of outlet streams. In some cases, the solids comprise one or more cells.

[0022] In another aspect, provided herein is a method for distributing a fluid, comprising: (a) providing a primary fluid distribution device and a plurality of secondary fluid distribution devices fluidically connected to the primary fluid distribution device, wherein the primary fluid distribution device is configured to distribute the fluid between each secondary fluid distribution device; (b) directing a fluid to the primary fluid distribution device, thereby allowing the fluid to flow in a substantially vertical direction from the primary fluid distribution device through the plurality of secondary fluid distribution devices; and (c) producing a plurality of output streams, wherein the volume of the fluid in each outlet stream is within 10% of a mean volume of the plurality of outlet stream

[0023] In some cases, the fluid flows from the primary fluid distribution device to the plurality of secondary fluid distribution devices through tubing. In some cases, the method further comprises releasing air from the primary fluid distribution device or the plurality of secondary fluid distribution devices through one or more vents. In some cases, the primary fluid distribution device or the plurality of secondary fluid distribution is comprised of an optically transparent material. In some cases, the method further comprises optically observing a flow pattern of the fluid through the optically transparent material.

[0024] In some cases, the fluid comprises solids. In some cases, a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of the plurality of outlet streams. In some cases, a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of the plurality of outlet streams. In some cases, the solids comprise one or more cells.

[0025] In some cases, the method further comprises collecting an outlet stream of the plurality of outlet streams in a container, wherein when in the container, the output stream is not in fluid communication with another outlet stream of the plurality of outlet streams. In some cases, the container comprises a well of a well-plate. In some cases, the container comprises a test tube.

[0026] In another aspect, provided herein is a non-transitory computer readable medium comprising machine-executable code that, upon execution by one or more computer processors, implements a method for distributing a fluid, the method comprising: (a) directing a fluid to an input channel of a flow distribution device, wherein the flow distribution device comprises the input channel and a plurality of secondary channels, wherein each secondary channel of the plurality of secondary channels is fluidically connected to the input channel via a flow splitter; (b) allowing the fluid to flow through the flow splitter and the plurality of second channels; and (c) producing a plurality of output streams, wherein the volume of the fluid in each outlet stream is within 10% of a mean volume of the plurality of outlet streams.

[0027] Another aspect of the present disclosure provides a non-transitory computer readable medium comprising machine executable code that, upon execution by one or more computer processors, implements any of the methods above or elsewhere herein.

[0028] Another aspect of the present disclosure provides a system comprising one or more computer processors and computer memory coupled thereto. The computer memory comprises machine executable code that, upon execution by the one or more computer processors, implements any of the methods above or elsewhere herein.

[0029] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.

INCORPORATION BY REFERENCE

[0030] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such contradictory material.

BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0032] FIG. 1A illustrates a three-dimensional rendering of a 16-output vertical fluid flowdistribution device, according to some embodiments.

[0033] FIG. IB illustrates a three-dimensional rendering of the channels of a 16-output vertical fluid flow-distribution device, according to some embodiments.

[0034] FIG. 2A illustrates a front view of the vertical-flow fluid distribution device of FIG. 1, according to some embodiments.

[0035] FIG. 2B illustrates a side view of the vertical-flow fluid distribution device of FIG. 1, according to some embodiments.

[0036] FIG. 3A illustrates a front view of a vertical-flow fluid distribution device of FIG. 1 along with relative dimensions, according to some embodiments

[0037] FIG. 3B illustrates a side view of a vertical-flow fluid distribution device of FIG. 1 along with relative dimensions, according to some embodiments.

[0038] FIG. 4A illustrates a top-view of the cross section labeled D of the vertical-flow fluid distribution illustrated in FIG. 2B, according to some embodiments.

[0039] FIG. 4B illustrates a top-view of the cross section labeled E of the vertical-flow fluid distribution illustrated in FIG. 3B, according to some embodiments.

[0040] FIG. 4C illustrates a top-view of the cross section labeled F of the vertical-flow fluid distribution illustrated in FIG. 2B, according to some embodiments.

[0041] FIG. 4D illustrates a top-view of the cross section labeled G of the vertical-flow fluid distribution illustrated in FIG. 3B, according to some embodiments.

[0042] FIG. 4E illustrates a top-view of the cross section labeled H of the vertical-flow fluid distribution illustrated in FIG. 2B, according to some embodiments.

[0043] FIG. 5A is a photograph of a 16-output vertical -flow fluid distribution device made of an optically transparent material, according to some embodiments.

[0044] FIG. 5B is a front- view photograph of a 16-output vertical -flow fluid distribution device made of an optically transparent material, according to some embodiments.

[0045] FIG. 5C is a photograph of the 16 outputs on the bottom of a 16-output vertical -flow fluid distribution device, according to some embodiments.

[0046] FIG. 6A illustrates a three-dimensional rendering of a 4-output vertical-flow fluid distribution device, according to some embodiments.

[0047] FIG. 6B is a photograph of a 4-output vertical-flow fluid distribution device, according to some embodiments.

[0048] FIG. 7A illustrates a front view of a four-stream outlet vertical-flow fluid distribution device along with relative dimensions, according to some embodiments.

[0049] FIG. 7B illustrates a side view of a four-stream outlet vertical-flow fluid distribution device along with relative dimensions, according to some embodiments.

[0050] FIG. 7C illustrates a top-view of the cross section labeled A of the vertical-flow fluid distribution illustrated in FIG. 7A, according to some embodiments.

[0051] FIG. 7D illustrates a top-view of the cross section labeled B of the vertical-flow fluid distribution illustrated in FIG. 7A, according to some embodiments.

[0052] FIG. 8 illustrates a front view of a system comprising four 16-stream outlet vertical-flow distribution devices, according to some embodiments.

[0053] FIG. 9 illustrates an alternative view of the vertical -flow distribution system of FIG. 8, according to some embodiments.

[0054] FIG. 10 illustrates the vertical-flow distribution system of FIG. 8 without any tubing, according to some embodiments.

[0055] FIG. 11 illustrates a herringbone structure that could be used on the inside of a flow channel, according to some embodiments.

[0056] FIG. 12 illustrates a 16-output fluid distribution device with one vent on all four sides, according to some embodiments.

[0057] FIG. 13 shows a box plot of the volume distribution of a fluid after flowing through a 16- output fluid distribution device, according to some embodiments.

[0058] FIG. 14 shows a box plot of the volume distribution of a fluid after flowing through two joined 16-output fluid distribution devices, according to some embodiments.

[0059] FIG. 15 shows a box plot of cell distribution of a fluid after flowing through a 16-output fluid distribution device, according to some embodiments.

[0060] FIG. 16A shows a bar graph of the volume distribution of a fluid after flowing through a 16- output fluid distribution device (Rig A), according to some embodiments.

[0061] FIG. 16B shows a bar graph of the cell distribution of a fluid after flowing through a 16- output fluid distribution device (Rig A), according to some embodiments.

[0062] FIG. 16C shows a bar graph of the volume distribution of a fluid after flowing through a 16- output fluid distribution device (Rig B), according to some embodiments.

[0063] FIG. 16D shows a bar graph of the cell distribution of a fluid after flowing through a 16- output fluid distribution device (Rig B), according to some embodiments.

[0064] FIG. 17 shows a box plot of the volume distribution of a fluid after flowing through a 4- output fluid distribution device, according to some embodiments.

[0065] FIG. 18 shows a box plot of particle distribution of a fluid after flowing through a 4-output fluid distribution device, according to some embodiments.

[0066] FIG. 19 shows a box plot of cell distribution of a fluid after flowing through a 4-output fluid distribution device, according to some embodiments.

[0067] FIG. 20 schematically illustrates a computer system that is programmed or otherwise configured to implement methods provided herein.

[0068] FIG. 21A shows filling time as a function of the number of outputs of a fluid distribution device for devices with a constant head pressure and 3, 4, 16, 32, and 64 outputs, according to some embodiments. [0069] FIG. 21B shows filling time as a function of the number of outputs of a fluid distribution device for devices with a constant head pressure greater than 64 outputs, according to some embodiments.

[0070] FIG. 22 A illustrates a three-dimensional rendering of a 3 -output vertical fluid distribution device, according to some embodiments.

[0071] FIG. 22B is a photograph of a 3 -output vertical-flow fluid distribution device, according to some embodiments.

[0072] FIG. 23 shows a box plot of the volume distribution of a fluid after flowing through a 3- output fluid distribution device, according to some embodiments.

[0073] FIG. 24 shows the average cell viability of Jurkat cells at the output of a 16-output vertical fluid distribution device.

[0074] FIG. 25A shows the linear scaling of pressure as a function of the number of device outputs, according to some embodiments.

[0075] FIG. 25B shows filling time as a function of the number of outputs of fluid distribution devices with scaled pressure, according to some embodiments.

DETAILED DESCRIPTION

[0076] While various embodiments have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions can occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the disclosure described herein can be employed.

[0077] Whenever the term “about,” “at least,” “greater than,” or “greater than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “about,” “at least,” “greater than” or “greater than or equal to” applies to each of the numerical values in that series of numerical values. For example, greater than or equal to 1, 2, or 3 is equivalent to greater than or equal to 1, greater than or equal to 2, or greater than or equal to 3.

[0078] Whenever the term “no more than,” “less than,” or “less than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “no more than,” “less than,” or “less than or equal to” applies to each of the numerical values in that series of numerical values. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1.

Overview

[0079] The present disclosure provides devices, systems, and method for dividing or distributing a total amount of fluid into equal aliquots. For fluids comprising a total amount of solids, the devices, systems, and methods described herein can divide the total amount of solids into equal aliquots with the correct concentration and an equal number of solids in each output. The system can comprise one or more fluid channels and flow splitters configured to divide fluid evenly between a plurality of output channels.

Vertical- Flow Distribution Device

[0080] A vertical-fluid flow distribution device can be used to distribute a volume of fluid between one or more output streams. The fluid can comprise a liquid or an aqueous solution. The fluid can comprise one or several analytes. The fluid can comprise solid particles. The solid particles can be present in the fluid at a given concentration. The solid particles can comprise one or more biological components. The biological component can comprise a cell. In some embodiments, the cell can comprise a eukaryotic cell, a prokaryotic cell, a fungal cell, a protozoan, an algal cell, a plant cell, an animal cell (e.g., a human cell), or any other suitable cell. The biological component can comprise a virus, a bacterium, a nucleic acid (e.g., DNA, or RNA), a protein, or a combination thereof. The fluid can comprise a reagent. A reagent can comprise, for example, balanced salt solutions, buffers, detergents, chelators, or any materials or substances used in chemical analysis, biological analysis, or in other reactions. Any operation using said reagents can be carried out in a closed environment (no opening of the device at any time). The fluid can be cell culture medium.

[0081] In some cases, the fluid comprises solids with a particle diameter of about 1 pm to about 100 pm. In some cases, the fluid comprises solids with a particle diameter of about 1 pm to about 5 pm, about 1 pm to about 10 pm, about 1 pm to about 25 pm, about 1 pm to about 50 pm, about 1 pm to about 75 pm, about 1 pm to about 100 pm, about 5 pm to about 10 pm, about 5 pm to about 25 pm, about 5 pm to about 50 pm, about 5 pm to about 75 pm, about 5 pm to about 100 pm, about 10 pm to about 25 pm, about 10 pm to about 50 pm, about 10 pm to about 75 pm, about 10 pm to about 100 pm, about 25 pm to about 50 pm, about 25 pm to about 75 pm, about 25 pm to about 100 pm, about 50 pm to about 75 pm, about 50 pm to about 100 pm, or about 75 pm to about 100 pm. In some cases, the fluid comprises solids with a particle diameter of about 1 pm, about 5 pm, about 10 pm, about 25 pm, about 50 pm, about 75 pm, or about 100 pm. In some cases, the fluid comprises solids with a particle diameter of at least about 1 pm, about 5 pm, about 10 pm, about 25 pm, about 50 pm, or about 75 pm. In some cases, the fluid comprises solids with a particle diameter of at most about 5 pm, about 10 pm, about 25 pm, about 50 pm, about 75 pm, about 100 pm, about 200 pm, about 300pm, about 400 pm, about 500pm or about 600pm.

[0082] A vertical-fluid flow distribution device can comprise an input channel configured to receive a fluid. In some cases, the input channel is fluidically connected to one or more levels of branched channels. The branched channels can be located below the input channel such that gravity at least partially facilitates flow of the fluid from the input channel through the branched channels.

[0083] In some cases, a fluid distribution device can include an inline staggered mixer. An inline staggered mixer can be useful to counteract preferential distribution of solids when flowed through a split point. In some cases, the inline staggered mixer lines the inner surface of a channel (including both an input channel or branching channel). The mixer can have a raised structure or pattern configured to homogenize flow of a fluid through that channel. This raised structure can be laid out in a herringbone pattern. FIG. 11 illustrates a schematic of a herringbone pattern which can coat an inner surface or portion thereof of any fluidic channels described herein.

[0084] A fluid distribution device can be used to distribute a volume of fluid into multiple outlet streams. The given volume of fluid can take a specific amount of time to flow through the fluid distribution device and be distributed into one or more outlet streams. For example, distribution of 5 liters of cell culture medium comprising 2x10⁹ cells/mL into 256 vials can take approximately 3 minutes (at a given constant pressure).

[0085] In some cases, the fluid flows through the fluid distribution device at a volumetric flow rate of about 1 mL/s to about 20 mL/s. In some cases, the fluid flows through the fluid distribution device at a volumetric flow rate of about 1 mL/s to about 5 mL/s, about 1 mL/s to about 7.5 mL/s, about 1 mL/s to about 9 mL/s, about 1 mL/s to about 10 mL/s, about 1 mL/s to about 11 mL/s, about 1 mL/s to about 12.5 mL/s, about 1 mL/s to about 15 mL/s, about 1 mL/s to about 20 mL/s, about 5 mL/s to about 7.5 mL/s, about 5 mL/s to about 9 mL/s, about 5 mL/s to about 10 mL/s, about 5 mL/s to about 11 mL/s, about 5 mL/s to about 12.5 mL/s, about 5 mL/s to about 15 mL/s, about 5 mL/s to about 20 mL/s, about 7.5 mL/s to about 9 mL/s, about 7.5 mL/s to about 10 mL/s, about 7.5 mL/s to about 11 mL/s, about 7.5 mL/s to about 12.5 mL/s, about 7.5 mL/s to about 15 mL/s, about 7.5 mL/s to about 20 mL/s, about 9 mL/s to about 10 mL/s, about 9 mL/s to about 11 mL/s, about 9 mL/s to about 12.5 mL/s, about 9 mL/s to about 15 mL/s, about 9 mL/s to about 20 mL/s, about 10 mL/s to about 11 mL/s, about 10 mL/s to about 12.5 mL/s, about 10 mL/s to about 15 mL/s, about 10 mL/s to about 20 mL/s, about 11 mL/s to about 12.5 mL/s, about 11 mL/s to about 15 mL/s, about 11 mL/s to about 20 mL/s, about 12.5 mL/s to about 15 mL/s, about 12.5 mL/s to about 20 mL/s, or about 15 mL/s to about 20 mL/s. In some cases, the fluid flows through the fluid distribution device at a volumetric flow rate of about 1 mL/s, about 5 mL/s, about 7.5 mL/s, about 9 mL/s, about 10 mL/s, about 11 mL/s, about 12.5 mL/s, about 15 mL/s, or about 20 mL/s. In some cases, the fluid flows through the fluid distribution device at a volumetric flow rate of at least about 1 mL/s, about 5 mL/s, about 7.5 mL/s, about 9 mL/s, about 10 mL/s, about 11 mL/s, about 12.5 mL/s, or about 15 mL/s. In some cases, the fluid flows through the fluid distribution device at a volumetric flow rate of at most about 5 mL/s, about 7.5 mL/s, about 9 mL/s, about 10 mL/s, about 11 mL/s, about 12.5 mL/s, about 15 mL/s, or about 20 mL/s. In some cases, the channel dimensions are chosen so that the Re number stays constant through the device and below 2300, e.g., in a laminar regime of operation.

[0086] After a fluid flows through one or more levels of branched channels, it can exit the fluid distribution system through a plurality of outputs. In some cases, the outputs are fluidically connected to the one or more levels of branched channels. The plurality of outputs can be located below the one or more levels of branched channels such that gravity at least partially facilitates flow of the fluid from the branched channels through the plurality of outputs. In some cases, the fluid exits the outputs as a plurality of outlet streams.

[0087] The volume of fluid in each output stream of the fluid distribution system can be roughly equal. In some cases, the volume of fluid in each output is within 5% of the mean volume of all outlet streams. In some cases, the volume of fluid in each output is within 0% to 10% of the mean volume of all outlet streams. In some cases, the volume of fluid in each output is within 1% to 2%, 1% to 3%, 1% to 4%, 1% to 4.5%, 1% to5 %, 1% to 5.5%, 1% to 6%, 1% to 7%, 1% to 8%, 1% to 9%, 1% to 10%, 2% to 3%, 2% to 4%, 2% to 4.5%, 2% to 5%, 2% to 5.5%, 2% to 6%, 2% to 7%, 2% to 8%, 2% to 9%, 2% to 10%, 3% to 4%, 3% to 4.5%, 3% to 5%, 3 % to 5.5%, 3% to 6%, 3% to 7%, 3% to 8%, 3% to 9%, 3% to 10%, 4% to 4.5%, 4% to 5%, 4% to 5.5%, 4% to 6%, 4% to 7%, 4% to 8%, 4% to 9%, 4% to 10%, 4.5% to 5%, 4.5% to 5.5%, 4.5% to 6%, 4.5% to 7%, 4.5% to 8%, 4.5% to 9%, 4.5% to 10%, 5% to 5.5%, 5% to 6%, 5% to 7%, 5% to 8%, 5% to 9%, 5% to 10%, 5.5% to 6%, 5.5% to 7%, 5.5% to 8%, 5.5% to 9%, 5.5% to 10%, 6% to 7%, 6% to 8%, 6% to 9%, 6% to 10%, 7% to 8%, 7% to 9%, 7% to 10%, 8% to 9%, 8% to 10%, or 9% to 10% of the mean volume of all outlet streams. In some cases, the volume of fluid in each output is within 1%, 2%, 3%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, or 10% of the mean volume of all outlet streams. In some cases, the volume of fluid in each output is within at least 1%, 2%, 3%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, or 9% of the mean volume of all outlet streams. In some cases, the volume of fluid in each output is within at most 2%, 3%, 4%, 4.5%, 5%, 5.5%, 6%, 7%, 8%, 9%, or 10%. of the mean volume of all outlet streams.

[0088] The concentration of solids in each output stream of the fluid distribution system can be roughly equal. In some cases, the concentration of solids in each outlet stream is within 10% of the mean concentration of solids of all outlet streams. In some cases, the concentration of solids in each outlet stream is within 0% to 10% of the mean concentration of solids of all outlet streams. In some cases, the concentration of solids in each outlet stream is within 5% to 8%, 5% to 9%, 5% to 9.5%, 5% to 10%, 5% to 10.5%, 5% to 11%, 5% to 12%, 5% to 15%, 8% to 9%, 8% to 9.5%, 8% to 10%, 8% to 10.5%, 8% to 11%, 8% to 12%, 8% to 15%, 9% to 9.5%, 9% to 10%, 9% to 10.5%, 9% to 11%, 9% to 12%, 9% to 15%, 9.5% to 10%, 9.5% to 10.5%, 9.5% to 11%, 9.5% to 12%, 9.5% to 15%, 10% to 10.5%, 10% to 11%, 10% to 12%, 10% to 15%, 10.5% to 11%, 10.5% to 12%, 10.5% to 15%, 11% to 12%, 11% to 15%, or 12% to 15% of the mean concentration of solids of all outlet streams. In some cases, the concentration of solids in each outlet stream is within 5%, 8%, 9%, 9.5%, 10%, 10.5%, 11%, 12%, or 15% of the mean concentration of solids of all outlet streams. In some cases, the concentration of solids in each outlet stream is within at least 5%, 8%, 9%, 9.5%, 10%, 10.5%, 11%, or 12% of the mean concentration of solids of all outlet streams. In some cases, the concentration of solids in each outlet stream is within at most 8%, 9%, 9.5%, 10%, 10.5%, 11%,

12%, or 15% of the mean concentration of solids of all outlet streams. [0089] A vertical-flow fluid distribution device can have a plurality of output streams. In some cases, a fluid distribution device can have 2 outlet streams to 1024 outlet streams. In some cases, a fluid distribution device can have 2 outlet streams to 4 outlet streams, 2 outlet streams to 8 outlet streams, 2 outlet streams to 16 outlet streams, 2 outlet streams to 32 outlet streams, 2 outlet streams to 64 outlet streams, 2 outlet streams to 128 outlet streams, 2 outlet streams to 256 outlet streams to 512 outlet streams to 1024 outlet streams, 4 outlet streams to 8 outlet streams, 4 outlet streams to 16 outlet streams, 4 outlet streams to 32 outlet streams, 4 outlet streams to 64 outlet streams, 4 outlet streams to 128 outlet streams, 4 outlet streams to 256 outlet streams, 8 outlet streams to 16 outlet streams, 8 outlet streams to 32 outlet streams, 8 outlet streams to 64 outlet streams, 8 outlet streams to 128 outlet streams, 8 outlet streams to 256 outlet streams, 16 outlet streams to 32 outlet streams, 16 outlet streams to 64 outlet streams, 16 outlet streams to 128 outlet streams, 16 outlet streams to 256 outlet streams, 32 outlet streams to 64 outlet streams, 32 outlet streams to 128 outlet streams, 32 outlet streams to 256 outlet streams, 64 outlet streams to 128 outlet streams, 64 outlet streams to 256 outlet streams, 128 outlet streams to 256 outlet streams, 128 outlet streams to 512 outlet streams, 128 outlet streams to 1024 outlet streams. In some cases, a fluid distribution device can have 2 outlet streams, 4 outlet streams, 8 outlet streams, 16 outlet streams, 32 outlet streams, 64 outlet streams, 128 outlet streams, 256 outlet streams, 512 outlet streams, or 1024 outlet streams. In some cases, a fluid distribution device can have at least 2 outlet streams, 4 outlet streams, 8 outlet streams, 16 outlet streams, 32 outlet streams, 64 outlet streams, 128 outlet streams, 256, outlet streams, 512 outlet streams, or 1024 outlet streams. In some cases, a fluid distribution device can have at most 4 outlet streams, 8 outlet streams, 16 outlet streams, 32 outlet streams, 64 outlet streams, 128 outlet streams, 256 outlet streams, 512 outlet streams, or 1024 outlet streams. FIG. 5C illustrates a bottom view of a 16-output fluid distribution device and shows that there are 16 equally sized and spaced outputs. [0090] A channel level can be separated from an upper channel directly above by a split point. In some cases, the split point can be a binary split point. In some cases, the split point can divide an input stream into more than two channels. In some cases, the split point is a three-way split point, a four-way split point, a five way-split point, or a six-way split point. The split point can result in any number of branches. A split point can be configured to distribute fluid roughly equally between all lower branching channels. A binary split can be configured to split an upper channel into two lower channels. A binary split point can be configured to distribute fluid from an upper channel between two lower channels. A binary split point can comprise an upper channel and two branched channels. In some cases, the two branched channels below a binary split point are spaced 180° apart.

[0091] A three-way split can be configured to split an upper channel into three lower channels. A three-way split point can be configured to distribute fluid from an upper channel between three lower channels. In some cases, the three branched channels below a three-way split point are spaced 120° apart.

[0092] A four-way split can be configured to split an upper channel into four lower channels. A four-way split point can be configured to distribute fluid from an upper channel between four lower channels. In some cases, the four branched channels below a four-way split point are spaced 90° apart.

[0093] A branched channel can slope downward from the upper channel by a certain angle. In some cases, the angle between an upper channel and a branched channel can be about 100° to about 140°. In some cases, the angle between an upper channel and a branched channel can be about 100° to about 105°, about 100° to about 110°, about 100° to about 115°, about 100° to about 120°, about 100° to about 125°, about 100° to about 130°, about 100° to about 135°, about 100° to about 140°, about 105° to about 110°, about 105° to about 115°, about 105° to about 120°, about 105° to about

125°, about 105° to about 130°, about 105° to about 135°, about 105° to about 140°, about 110° to about 115°, about 110° to about 120°, about 110° to about 125°, about 110° to about 130°, about 110° to about 135°, about 110° to about 140°, about 115° to about 120°, about 115° to about 125°, about 115° to about 130°, about 115° to about 135°, about 115° to about 140°, about 120° to about 125°, about 120° to about 130°, about 120° to about 135°, about 120° to about 140°, about 125° to about 130°, about 125° to about 135°, about 125° to about 140°, about 130° to about 135°, about 130° to about 140°, or about 135° to about 140°. In some cases, the angle between an upper channel and a branched channel can be about 100°, about 105°, about 110°, about 115°, about 120°, about 125°, about 130°, about 135°, or about 140°. In some cases, the angle between an upper channel and a branched channel can be at least about 100°, about 105°, about 110°, about 115°, about 120°, about 125°, about 130°, or about 135°. In some cases, the angle between an upper channel and a branched channel can be at most about 105°, about 110°, about 115°, about 120°, about 125°, about 130°, about 135°, or about 140°. In some cases, all binary splits in a fluid distribution system can have the angle between the upper channel and a branched channel.

[0094] A vertical-flow fluid distribution device can comprise an optically transparent material. The device can be fabricated in optically transparent material or a combination of different types of materials. Such materials can be transparent so that imaging technology can be coupled. For example, the optically transparent materials can comprise polymers, acrylics, silicones, ceramics, glass, quartz, resin, clear resin, biocompatible resins, including low viscosity biocompatible resins. The material should be sterilizable via standard sterilization techniques such as but not limited to autoclave, gamma irradiation, ethylene oxide treatment, etc. The material can be selected based on its optimum low viscosity, mechanical properties (material strength) and/or UV resistance. In some cases, the fluid distribution or device can be fabricated using a three-dimensional printer. In some cases, the fluid distribution device can be made of a material compatible with dimethyl sulfoxide

(DMSO). A DMSO-compatible material ensures that DMSO can be mixed in the fluid so that the output containers can be cryogenically preserved after splitting. In some cases, the fluid distribution device is sterile. FIGs. 5A and 5B illustrate a 16-output fluid distribution device made of an optically transparent material. FIG. 6B illustrates a 4-output fluid distribution device made of an optically transparent material.

[0095] A vertical-flow fluid distribution device can include one or more vents. Vents can be used to push out excess air and stabilize fluid flow, which provides less variation of fluid outputs. In some cases, a fluid distribution device can have vents on each side. In some cases, a fluid distribution device has one, two, three, or four vents. FIG. 12 shows a 16-output fluid distribution device with one vent on all four sides. The vents can be fitted with a minimum of 0.2pm aseptic filter to maintain a closed sterile environment.

[0096] In some cases, one or more channels have a curved profile. A curved surface on the branching zone of a channel can influence the flow velocity profile of the vertical fluid distribution device. In some cases, a curved surface reduces dead zones which can reduce localized high pressure build-up during flow. A curved channel or curved branching zone can result in a smoother flow regime. A curved channel or curved branching zone can reduce the pressure drop through the channel, which can reduce the shear stress acting upon the cells during splitting.

Design and Fabrication of a Fluid Distribution Device

[0097] A set of modular units (similar to the fluid distribution devices shown in FIG. 1A and IB) can be designed to split fluid flow from a single inlet into 16 outlets (1 x16) and to accommodate the footprint of the collection vials. The collection vials can be standard filling vessels such as but not limited to centrifuge tubes (e.g. five milliliter or fifty milliliter volumes); cryo-bags or freezing bags (lOmL or 50mL or 250mL or 500mL or 750mL or lOOOmL volume capacity); glass injection vials or freeze drying vials (lOmL to lOOmL volume) all fitted with aseptic connectors Push-in connectors can be integrated at the inlet of the device (which can have a diameter of, for example, 8 mm as shown in FIG. 3B and 4A) and at the outlets of the device (each which can have a diameter of, for example, 3.5 mm as shown in FIG. 4B). Furthermore, to prevent the introduction of unnecessary constrictions in the flow path, any feed-in tubing can be selected to match the inner diameter of an input channel. The channel cross-section can be square. Therefore, the input channel can have a hydraulic diameter (DH = 4-A/P) of 3.5 mm as shown in FIG. 4A. To allow a developed parabolic flow, a minimal channel length of approximately 10 times the hydraulic diameter (DH) can be considered. At the end of that recommended length, a bifurcation split can be introduced to divide the channel in the middle, creating two identical channels with a rectangular cross-section (H:3,5 mm, W: 1.75 mm, as shown in FIG. 4B). After the bifurcation, the two resulting channels can be split further whilst applying the same design rules; namely, channel length, which is ten times the new hydraulic diameter, channel height (Hi: 1.75 mm), channel width (Wi:0.85 mm), and bifurcation angle of 120°. Thus, the aspect ratio for each channel can remain constant (2: 1, H:W); in contrast to the main channel (1: 1). Four bifurcations yield a split of one into sixteen (1 2⁴) outlets, which is shown in the channel network depicted in FIG. IB. For a fluid distribution device consisting of one input channel and a series of bifurcated splits, the number of outlets can be calculated as (lx2^x), where x is the number of bifurcations. For example, a fluid distribution device with one input channel and one bifurcation can result in two outputs. A fluid distribution device with one input channel and 5 bifurcations can result in 32 outputs. In another embodiment, the fluid distribution device can result to an odd number of outputs (e.g. 1 input to 3 outputs).

[0098] The fluid distribution devices described herein can be manufactured using a three- dimensional printer.

[0099] The external dimensions, connection ports, and channel outlets can be measured using a calliper and compared to the initial designs. For built-in features, such as an in-line 3D staggered herringbone mixer, the channel can be milled and inspected using an optical profiler such as the Wyko NT9100, Veeco, US.

16-Output Fluid Distribution Device

[00100] FIGs. 1A and IB illustrate one embodiment of a three-dimensional rendering of a vertical fluid distribution device with 16 outputs. The fluid distribution device 100 can comprise an input channel 105. The input channel can be configured to receive a volume of fluid. In some cases, the volume of fluid comprises solids at a given concentration. The fluid can enter the input channel 105 and flow in a substantially vertical direction through the fluid distribution device 100. The fluid can exit the input channel 105 and flow through a binary split 110. The binary split 110 can divide the input channel 105 into two secondary channels 115. The secondary channels 115 can be located across from each other in a first plane (x plane). Fluid can enter the secondary channels 115. Fluid through a secondary channel can flow in a substantially vertical direction towards a second binary split 120. The binary split 120 can divide the secondary channel 115 into two tertiary channels 125. The resulting tertiary channels 125 can be located across from each other in a second plane (y plane). This split can result in four total tertiary channels 125. The fluid can continue to flow through a series of binary splits and channels in a similar manner. In some cases, as shown in FIG. 1, the fluid distribution device will produce 16 total outputs 130. The fluid flowing from each output can be of substantially the same volume. The concentration of solids in each output can be substantially the same.

[00101] Referring to FIGs. 2A and 2B, fluid can enter the input channel 205 and flow in a substantially vertical direction through the fluid distribution device. Cross section D, shown in FIG. 4A, shows the top view of the input channel 205. The fluid can exit the input channel 205 and flow through a binary split 210. The binary split 210 can divide the input channel 205 into two secondary channels 215. The secondary channels 215 can be located across from each other in a first plane (x plane). Cross section E, shown in FIG. 4B, shows the top view of the device with two total channels 215. The fluid can flow though the secondary channel 215 and into a binary split 220. The binary split 220 can divide each secondary channel 215 into two tertiary channels 225. The tertiary channels 225 can be located across from each other in a first plane (y plane). Cross section F, shown in FIG. 4C, shows the top view of the device with four total channels 225. The fluid can flow through the tertiary channel 225 and into a binary split 230. The binary split 230 can divide each tertiary channel 225 into two quarternary channels 235. The tertiary channels 235 can be located across from each other in a first plane (x plane). Cross section G, shown in FIG. 4D, shows the top view of the device with eight total channels 235. The fluid can flow through the quarternary channel 235 and into a binary split 240. The binary split 240 can divide each quarternary channel 235 into two quinary channels 245. The quarternary channels 245 can be located across from each other in a first plane (y plane). Cross section H, shown in FIG. 4E, shows the top view of the device with sixteen total channels 245.

[00102] FIG. 3A illustrates a view of the vertical-flow fluid distribution device of FIG. 2A along a first plane (x plane). FIG. 3B illustrates a view of the vertical-flow fluid distribution device of FIG. 2B along a second plane (y plane). As shown in FIGs. 3A and 3B, the angle between an upper channel and a branched channel can be about 120° and can be identical in each binary split.

4-Output Fluid Distribution Device

[00103] FIGs. 6A and 6B illustrate a three-dimensional rendering and a photograph of a vertical fluid flow-distribution device with 4 outputs, respectively. Referring to FIGs. 7A-7D, fluid can enter the input channel 305 and flow in a substantially vertical direction through the fluid distribution device. The fluid can exit the input channel 305 and flow through a binary split 310. The binary split 310 can divide the input channel 305 into two secondary channels 315. The secondary channels 315 can be located across from each other in a first plane (y plane). Cross section A, shown in FIG. 7C, shows the top view of the device with two total channels 315. The fluid can flow though the secondary channel 315 and into a binary split 320. The binary split 320 can divide each secondary channel 315 into two tertiary channels 325. The tertiary channels 325 can be located across from each other in a second plane (x plane). Cross section B, shown in FIG. 7D, shows the top view of the device with four total channels 325.

3-Output Fluid Distribution Device

[00104] FIGs. 22A and 22B illustrate a three-dimensional rendering and a photograph of a vertical fluid flow-distribution device with 3 outputs, respectively. Referring to FIG. 22A, fluid can enter the input channel 405 and flow in a substantially vertical direction through the fluid distribution device. The fluid can exit the input channel 405 and flow through a three-way split 410. The three-way split 410 can divide the input channel 405 into three channels 415. The three-way split 410 can distribute the fluid evenly between the three output channels 415.

Vertical- Flow Fluid Distribution System

[00105] A vertical-flow fluid distribution system can be used to distribute a volume of fluid between one or more output streams. A fluid distribution system can comprise one or more fluid distribution devices as described herein. A vertical flow distribution device can comprise a primary fluid distribution device configured to receive a fluid. In some cases, the primary fluid distribution device is similar to the fluid distribution device illustrated in FIGs. 7A-7D. In some cases, the primary fluid distribution device can distribute 2, 4, 8, or 16 outlet streams.

[00106] The vertical flow distribution device can also comprise a plurality of secondary fluid distribution devices fluidically connected to the primary fluid distribution device. There can be 2, 4, 8, 16, or any number of secondary fluid distribution devices. The number of outlet streams distributed by the primary fluid distribution device can be equal to the number of secondary fluid distribution devices. In some cases, each primary fluid distribution outlet feeds into an inlet or input channel of a secondary fluid distribution device. In some cases, the primary fluid distribution device is fluidically connected to one or more secondary fluid distribution devices through a plurality of tubes. The primary fluid distribution device can be configured to distribute the fluid between each secondary fluid distribution device. In some cases, a secondary fluid distribution device is similar to the fluid distribution device illustrated in FIGs. 1A and IB. In some cases, each secondary fluid distribution device can distribute 2, 4, 8, or 16 outlet streams.

[00107] In some cases, a fluid distribution system can have 16 outlet streams to 1024 outlet streams. In some cases, a fluid distribution system can have 16 outlet streams to 32 outlet streams, 16 outlet streams to 64 outlet streams, 16 outlet streams to 128 outlet streams, 16 outlet streams to 256 outlet streams, 16 outlet streams to 512 outlet streams, 16 outlet streams to 128 outlet streams, 16 outlet streams to 640 outlet streams, 32 outlet streams to 64 outlet streams, 32 outlet streams to 128 outlet streams, 32 outlet streams to 256 outlet streams, 32 outlet streams to 512 outlet streams, 32 outlet streams to 128 outlet streams, 32 outlet streams to 640 outlet streams, 64 outlet streams to 128 outlet streams, 64 outlet streams to 256 outlet streams, 64 outlet streams to 512 outlet streams, 64 outlet streams to 128 outlet streams, 64 outlet streams to 640 outlet streams, 128 outlet streams to 256 outlet streams, 128 outlet streams to 512 outlet streams, 128 outlet streams to 128 outlet streams, 128 outlet streams to 640 outlet streams, 256 outlet streams to 512 outlet streams, 256 outlet streams to 128 outlet streams, 256 outlet streams to 1024 outlet streams, 512 outlet streams to 128 outlet streams, 512 outlet streams to 1024 outlet streams, or 128 outlet streams to 1024 outlet streams. In some cases, a fluid distribution system can have 16 outlet streams, 32 outlet streams, 64 outlet streams, 128 outlet streams, 256 outlet streams, 512 outlet streams, or 1024 outlet streams. In some cases, a fluid distribution system can have at least 16 outlet streams, 32 outlet streams, 64 outlet streams, 128 outlet streams, 256 outlet streams, 512 outlet streams, or 1024 outlet streams. In some cases, a fluid distribution system can have at most 32 outlet streams, 64 outlet streams, 128 outlet streams, 256 outlet streams, 512 outlet streams, or 1024 outlet streams.

[00108] The secondary fluid distribution devices can distribute outlet streams into a plurality of containers. In some cases, a secondary fluid distribution device outputs an outlet stream into an isolated container so that the outlet stream is not fluidically connected to any other outlet streams from other secondary fluid distribution devices. In some cases, the container is a well of a well-plate. In some cases, the container is a test tube. In some cases, the container is a centrifuge tube. In some cases, the container is cryo-bag or freeze-drying vial or injection vial.

[00109] In some cases, a primary fluid distribution device is elevated in height as compared to one or more secondary fluid distribution devices such that gravity at least partially facilitates flow of the fluid through the fluid distribution system. The secondary fluid distribution devices can be located at equal distances from each other within the fluid distribution system. Outlet streams can be located below the one or more levels of the fluid distribution system. In some cases, the outlet streams are located below the one or more secondary fluid distribution devices such that gravity at least partially facilitates flow of the fluid through the fluid distribution system. In some cases, the fluid distribution system is sterile. The fluid distribution system described herein can be closed, i.e., operations can be carried out in a closed environment (no opening of the system at any time).

64-Output Fluid Distribution System

Referring to FIGs. 8-10, a fluid distribution system can comprise one primary fluid distribution device (1). This primary distribution device is located in the center of the axis of fluid distribution that outputs 4 streams through tubing (19). The tubing (19) can feed the 4 streams into 4 secondary fluid distribution devices (2). Each secondary fluid distribution device (2) can output 16 streams through tubing (18). The tubing (18) can be fluidically connected to a centrifuge caddy (11). The centrifuge caddy (11) can be able to hold 16 centrifuge tubes (20). The tubing (18) can distribute the fluid into 64 separate centrifuge tubes. The volume of fluid in each centrifuge tube can be substantially the same. The concentration of solids in each output can be substantially the same. The fluid distribution system can include additional structural components as illustrated in FIG. 8. The system can include one or more plates (3), (4), (5), or (17). These plates can keep the fluid distribution devices oriented in a specific layout. The primary input is located on the center axis of the fluid distribution. In some cases, one or more plates (3), (4), or (5) are used to keep the secondary fluid distribution devices (2) equidistant from each other. The system can include one or more frame supports (6), (7), (8), (9), or (16). The system can include a base plate (10). The system can comprise a support plate (12). The system can include a docking plate (13). The docking plate (13) can be used to support the centrifuge caddy (11). Tube alignment structures (14) and (15) can be used to keep the centrifuge tubes (20) in a specific orientation. The tube alignment structures (14) and (15) can be used to keep the output vessels (centrifuge tubes or bags) equidistant from each other, i.e., to arrange the output vessels to a modular array while keeping minimum spatial footprint and specific orientation.

Applications

[00110] The systems and methods described herein can be used to distribute a cell culture medium after cells have undergone a series of bioprocessing steps and are washed and concentrated. Cells can be in their final container before infusion into a patient and suspended in a solution made from several reagents suitable for protecting the cells against cryo-storage and/or for infusing into the patient. For allogeneic products, where one batch is used for several patients, the batch of cells can be broadly distributed into n containers evenly, with the right concentration of reagents, with n ranging from <10 to several hundreds.

Computer Systems

[00111] In an aspect, the present disclosure provides computer systems that are programmed or otherwise configured to implement methods of the disclosure, e.g., any of the subject methods for fluid distribution. Computer systems can be used to automate any method described herein. FIG. 20 shows a computer system 2001 that is programmed or otherwise configured to implement a method for fluid distribution. The computer system 2001 can be configured to, for example, automate or control the amount of fluid that enters a fluid distribution device or system. The computer system 2001 can be configured to adjust a flow rate or an amount of fluid flow into or through a fluid distribution device or system, based on one or more user inputs or sensor readings. The computer system 2001 can be further configured to adjust the flow rate or an amount of fluid flow into or through a fluid distribution device or system in order to optimize (i.e., increase or decrease) the volume of fluid or concentration of solids in each outlet stream. The computer system 2001 can be an electronic device of a user or a computer system that is remotely located with respect to the electronic device. The electronic device can be a mobile electronic device.

[00112] The computer system 2001 can include a central processing unit (CPU, also "processor" and "computer processor" herein) 2005, which can be a single core or multi core processor, or a plurality of processors for parallel processing. The computer system 2001 also includes memory or memory location 2010 (e.g., random-access memory, read-only memory, flash memory), electronic storage unit 2015 (e.g., hard disk), communication interface 2020 (e.g., network adapter) for communicating with one or more other systems, and peripheral devices 2025, such as cache, other memory, data storage and/or electronic display adapters. The memory 2010, storage unit 2015, interface 2020 and peripheral devices 2025 are in communication with the CPU 2005 through a communication bus (solid lines), such as a motherboard. The storage unit 2015 can be a data storage unit (or data repository) for storing data. The computer system 2001 can be operatively coupled to a computer network ("network") 2030 with the aid of the communication interface 2020. The network 2030 can be the Internet, an internet and/or extranet, or an intranet and/or extranet that is in communication with the Internet. The network 2030 in some cases is a telecommunication and/or data network. The network 2030 can include one or more computer servers, which can enable distributed computing, such as cloud computing. The network 2030, in some cases with the aid of the computer system 2001, can implement a peer-to-peer network, which can enable devices coupled to the computer system 2001 to behave as a client or a server.

[00113] The CPU 2005 can execute a sequence of machine-readable instructions, which can be embodied in a program or software. The instructions can be stored in a memory location, such as the memory 2010. The instructions can be directed to the CPU 2005, which can subsequently program or otherwise configure the CPU 2005 to implement methods of the present disclosure. Examples of operations performed by the CPU 2005 can include fetch, decode, execute, and writeback.

[00114] The CPU 2005 can be part of a circuit, such as an integrated circuit. One or more other components of the system 2001 can be included in the circuit. In some cases, the circuit is an application specific integrated circuit (ASIC).

[00115] The storage unit 2015 can store files, such as drivers, libraries and saved programs. The storage unit 2015 can store user data, e.g., user preferences and user programs. The computer system 2001 in some cases can include one or more additional data storage units that are located external to the computer system 2001 (e.g., on a remote server that is in communication with the computer system 2001 through an intranet or the Internet).

[00116] The computer system 2001 can communicate with one or more remote computer systems through the network 2030. For instance, the computer system 2001 can communicate with a remote computer system of a user (e.g., an operator managing or monitoring the bioprocessing). Examples of remote computer systems include personal computers (e.g., portable PC), slate or tablet PC's (e.g., Apple® iPad, Samsung® Galaxy Tab), telephones, Smart phones (e.g., Apple® iPhone, Android- enabled device, Blackberry®), or personal digital assistants. The user can access the computer system 2001 via the network 2030. [00117] Methods as described herein can be implemented by way of machine (e.g., computer processor) executable code stored on an electronic storage location of the computer system 2001, such as, for example, on the memory 2010 or electronic storage unit 2015. The machine executable or machine readable code can be provided in the form of software. During use, the code can be executed by the processor 2005. In some cases, the code can be retrieved from the storage unit 2015 and stored on the memory 2010 for ready access by the processor 2005. In some situations, the electronic storage unit 2015 can be precluded, and machine-executable instructions are stored on memory 2010.

[00118] The code can be pre-compiled and configured for use with a machine having a processor adapted to execute the code, or can be compiled during runtime. The code can be supplied in a programming language that can be selected to enable the code to execute in a pre-compiled or as- compiled fashion.

[00119] Aspects of the systems and methods provided herein, such as the computer system 2001, can be embodied in programming. Various aspects of the technology can be thought of as "products" or "articles of manufacture" typically in the form of machine (or processor) executable code and/or associated data that is carried on or embodied in a type of machine readable medium. Machineexecutable code can be stored on an electronic storage unit, such as memory (e.g., read-only memory, random-access memory, flash memory) or a hard disk. "Storage" type media can include any or all of the tangible memory of the computers, processors or the like, or associated modules thereof, such as various semiconductor memories, tape drives, disk drives and the like, which can provide non-transitory storage at any time for the software programming. All or portions of the software can at times be communicated through the Internet or various other telecommunication networks. Such communications, for example, can enable loading of the software from one computer or processor into another, for example, from a management server or host computer into the computer platform of an application server. Thus, another type of media that can bear the software elements includes optical, electrical and electromagnetic waves, such as used across physical interfaces between local devices, through wired and optical landline networks and over various airlinks. The physical elements that carry such waves, such as wired or wireless links, optical links or the like, also can be considered as media bearing the software. As used herein, unless restricted to non-transitory, tangible "storage" media, terms such as computer or machine "readable medium" refer to any medium that participates in providing instructions to a processor for execution.

[00120] Hence, a machine readable medium, such as computer-executable code, can take many forms, including but not limited to, a tangible storage medium, a carrier wave medium or physical transmission medium. Non-volatile storage media including, for example, optical or magnetic disks, or any storage devices in any computer(s) or the like, can be used to implement the databases, etc. shown in the drawings. Volatile storage media include dynamic memory, such as main memory of such a computer platform. Tangible transmission media include coaxial cables; copper wire and fiber optics, including the wires that comprise a bus within a computer system. Carrier-wave transmission media can take the form of electric or electromagnetic signals, or acoustic or light waves such as those generated during radio frequency (RF) and infrared (IR) data communications. Common forms of computer-readable media therefore include for example: a floppy disk, a flexible disk, hard disk, magnetic tape, any other magnetic medium, a CD-ROM, DVD or DVD-ROM, any other optical medium, punch cards paper tape, any other physical storage medium with patterns of holes, a RAM, a ROM, a PROM and EPROM, a FLASH-EPROM, any other memory chip or cartridge, a carrier wave transporting data or instructions, cables or links transporting such a carrier wave, or any other medium from which a computer can read programming code and/or data. Many of these forms of computer readable media can be involved in carrying one or more sequences of one or more instructions to a processor for execution. [00121] The computer system 2001 can include or be in communication with an electronic display

2035 that comprises a user interface (UI) 2040 for providing, for example, a portal for an operator to monitor or track one or more steps or operations of the fluid distribution methods and systems described herein. The portal can be provided through an application programming interface (API). A user or entity can also interact with various elements in the portal via the UI. Examples of UI's include, without limitation, a graphical user interface (GUI) and web-based user interface.

[00122] Methods and systems of the present disclosure can be implemented by way of one or more algorithms. An algorithm can be implemented by way of software upon execution by the central processing unit 2005. For example, the algorithm can be configured to adjust a flow rate or an amount of fluid flow into or through a fluid distribution device or system, based on one or more sensor readings or user inputs. In some embodiments, the algorithm can be further configured to adjust the flow rate or an amount of fluid flow into or through a fluid distribution device or system in order to optimize (i.e., increase or decrease) the volume of fluid or concentration of solids in each outlet stream.

EXAMPLES

EXAMPLE 1: Volume Distribution of a 16-output Fluid Distribution Device

[00123] In this example, a pressurized glass vessel was used to control the head pressure of a 16- output fluid distribution device (like the device shown in FIG. 5A). A pressurized glass vessel was connected through rigid tubing (1/16” Teflon tube, Upchurch, US) and standard connectors to a pressure controller. A compressor supplied compressed air to the pressure controller at a maximum pressure of 2 bar. The bottle was filled with the testing fluid, then it was sealed with screw top, and the stop valve was closed shut. Tubing was connected from the pressurized vessel to the 16-output fluid distribution device and downstream to 16 collection tubes (15mL Falcon, or 50 mL Falcon tubes, Thermo Scientific, UK). Next the glass bottle was hung upside down and secured with a pair of chemistry clamps. A pressure head was created at a constant pressure of 3.5 psi above the volume of liquid. By waiting for 15 s, after the pressure controller was switched on, the pressure is allowed to reach stable value within the pressurized vessel. The valve was switched to open, and the volume of fluid was distributed through the 16-output fluid distribution device into the collection tubes. Depending on the starting volume, fluid was collected into either 14-mL falcon tubes or 50-mL falcon tubes. The volume of water was measured gravimetrically. Namely, each tube was weighted together with its corresponding water content. Then a dry, empty tube was weighted, and the water content was calculated based on the difference. The test was repeated five times to gain statistical significance of the volume distribution results (n=5). These tests were timed also to measure the average flow rate through the network of channels. FIG. 13 shows a box plot of the resulting volume distribution of deionized water in the collection tubes. Five repetitions of this experiment were completed.

EXAMPLE 2: Parallel Splitting Between Two 16-output Fluid Distribution Devices

[00124] In this example, two 16-output fluid distribution devices (without vents) were tested in parallel using a commercially available Y-j unction with the same base setting for pressure described under Example 1. The pressure was set to a constant value of 7 psi (2 3.5psi) because the pressure scales linearly with the number of units connected. A stop valve was placed upstream from the Y- junction to prevent the liquid from rushing into the splitters. Once open, the valve let water into the connective tubing and into the two 16-output fluid distribution devices. Fifty milliliter Falcon tubes were placed at each outlet for collection. Gravimetric analysis was performed. Independent tests were run several times for statistically significant data (n=5). FIG. 14 shows a box plot for the volume distribution for this example. The means and standard deviations were plotted for each independent run. No significant differences were observed between the two devices running in parallel, or the independent run between devices. EXAMPLE 3: Jurkat Cell Distribution of a 16-output Fluid Distribution Device

[00125] In this example, fluorescently labelled Jurkat cells in a high-density culture were flowed through a 16-output fluid distribution device (like the device shown in FIG. 5A). The fluid distribution device had vents. The pressure was set to a constant value of 3.5 psi. Tests have also been conducted on a range of pressure values between 3.5psi and lOpsi. Cells were counted from each outlet. FIG. 15 demonstrates nearly even distribution of cells at each outlet with average at around 900k cells/mL. Volume distributions were within the ±5% margin. Deviation of the cells from each outlet is within the initially requested ±10% cells/mL.

EXAMPLE 4: Jurkat Cell Distribution Between Two 16-output Fluid Distribution Devices [00126] In this example, a fresh solution of cells was flowed through two separate 16-output fluid distribution devices (Rig A and Rig B), resulting in 32 outputs in total. For each outlet, the volume of solution was measured and the cells were counted. FIGs. 16A-16D demonstrate nearly even distribution of volume and cells at each outlet with average at around 160k cells/mL. The total initial volume of the cell solution was about 200mL. Volume distributions were within the ±5% margin and cell distribution deviation from each outlet is also within the initially requested ±10% cells/mL (at 4.6- 5.3% deviation across both rigs).

EXAMPLE 5: Volume Distribution of a 4-output Fluid Distribution Device

[00127] In this example, deionized water was flowed through a 4-output fluid distribution device (like the device shown in FIG. 6 A and FIG. 6B). A pressure head was created at a constant pressure of 9 psi above a volume of deionized water. By waiting for 15 s, after the pressure controller was switched on, the pressure was allowed to reach stable value within the pressurized vessel. The valve was switched to open, and the volume of deionized water was distributed through a 4-output fluid distribution device into four 50-mL collection tubes. The volume of water was measured gravimetrically. The test was repeated three times to gain statistical significance of the volume distribution results (n=3). FIG. 17 shows a box plot of the resulting volume distribution of deionized water in the collection tubes.

EXAMPLE 6: Fluorescent Particle Distribution of a 4-output Fluid Distribution Device

[00128] In this example, a mixture of 0.2 mL of 10-micron fluorescent beads in total of 200 mL of deionized water was prepared. The mixture was flowed through a 4-output fluid distribution device (like the device shown in FIG. 6A and FIG. 6B) into four separate collection tubes. The resulting fluorescent particle distribution is shown in FIG. 18.

EXAMPLE 7: Cell Distribution of a 4-output Fluid Distribution Device

[00129] In this example, a fresh solution of cells was flowed through a 4-output fluid distribution device. For each outlet, the number of cells was counted. FIG. 19 demonstrates the cell distribution after four independent runs through the 4-output fluid distribution device. Three measurements were taken per channel.

EXAMPLE 8: Volume Distribution of a 3-output Fluid Distribution Device

[00130] In this example, fluid was flowed through a 3-output fluid distribution device (like the device shown in FIG. 22A and FIG. 22B). A pressure head was created at a constant pressure of 3.5 psi above a volume of fluid. By waiting for 15 s, after the pressure controller was switched on, the pressure was allowed to reach stable value within the pressurized vessel. The valve was switched to open, and the volume of fluid was distributed through a 3-output fluid distribution device into three collection tubes. The volume of water was measured gravimetrically. The test was repeated three times to gain statistical significance of the volume distribution results (n=3). FIG. 23 shows a box plot of the resulting volume distribution of fluid in the collection tubes.

EXAMPLE 9: Cell Viability of Jurkat Cells at Output of a Vertical Fluid Distribution Device [00131] In this example, Jurkat cells in a high-density culture (3 million cells per mL) were flowed through a 16-output fluid distribution device (like the device shown in FIG. 5A). The pressure was set to a constant value of around 7 psi. Average cell viability was measured in each of three runs. FIG. 24 demonstrates an average viability of greater than 95%. Therefore, a high flow rate and corresponding shear during cell splitting through the vertical fluid distribution device did not induce significant stress on the Jurkat cells.

EXAMPLE 10: Filling Time as a Function of Number of Outputs (Constant Pressure)

[00132] In this example, the average total filling time was measured as a function of the number of outputs of a fluid distribution device. A constant head pressure was used for each fluid distribution device. As shown in FIG. 21A and FIG. 21B, the filling time of this system can linearly increase with the number of output containers (given a constant pressure/flow rate). In some cases, it takes approximately 3 seconds to fill 4 outputs and about 3 minutes (180 seconds) to fill 256 outputs.

EXAMPLE 11: Filling Time as a Function of Number of Outputs (Scaled Pressure)

[00133] In this example, the average total filling time of different outputs was measured while utilizing a scaled head pressure, i.e. the pressure was adjusted at the input channel depending on the number of outputs. For example, a 16-output fluid distribution used 7psi (483 mbar) pressure and a 32-output fluid distribution device used 14 psi (966 mbar). The linear scaling of pressure as a function of the number of outputs is shown in FIG. 25A. Next, the average filling time was recorded for each fluid distribution device. The average filling time for each device was plotted in FIG. 25B. As shown in FIG. 25B, when the pressure is adjusted to scale with the number of outputs, the average filling time will eventually reach a plateau. For this reason, the fluid distribution devices described herein can fill more containers (more outputs) in a shorter amount of time than can be achieved with sequential filling. For example, 128 outputs can be filled in approximately 24 seconds (~10 mL per output), which is only 4 seconds longer than the time to fill 64 outputs.

[00134] While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. It is not intended that the disclosure be limited by the specific examples provided within the specification. While the disclosure has been described with reference to the aforementioned specification, the descriptions and illustrations of the embodiments herein are not meant to be construed in a limiting sense. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. Furthermore, it shall be understood that all aspects of the disclosure are not limited to the specific depictions, configurations or relative proportions set forth herein which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the disclosure described herein can be employed in practicing the disclosure. It is therefore contemplated that the disclosure shall also cover any such alternatives, modifications, variations or equivalents. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

[00135] The following is a non-exhaustive list of embodiments of the invention which may or may not be claimed:

1. A vertical-flow fluid distribution device, comprising:

(a) an input channel configured to receive a fluid;

(b) one or more levels of branched channels fluidically connected to said input channel, wherein said one or more levels of branched channels are below said input channel; and

(c) a plurality of outputs fluidically connected to said one or more levels of branched channels, wherein said plurality of outputs are below said one or more levels of branched channels, wherein said device is configured to allow a fluid to flow in a substantially vertical direction from said input channel, through said one or more levels of branched channels, and to said plurality of outputs, and wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 10% of a mean volume of said plurality of outputs.

2. The device of embodiment 1, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

3. The device of embodiment 2, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

4. The device of embodiment 3, wherein said angle is about 100° to about 140°.

5. The device of embodiment 3, wherein said angle is about 115° to about 125°.

6. The device of embodiment 1, further comprising one or more vents comprising a sterile filter.

7. The device of embodiment 1, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

8. The device of embodiment 7, wherein said raised structure comprises a herringbone pattern.

9. The device of embodiment 1, wherein said system comprises an optically transparent material.

10. The device of embodiment 1, wherein said fluid comprises solids.

11. The device of embodiment 10, wherein a concentration of solids in each output is within 15% of a mean concentration of solids of said plurality of outputs.

12. The device of embodiment 10, wherein a concentration of solids in each output is within 10% of a mean concentration of solids of said plurality of outputs.

13. The device of embodiment 10, wherein said solids comprise one or more cells. 14. The device of embodiment 1, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 5% of a mean volume of said plurality of outputs.

15. A vertical-flow fluid distribution device, comprising:

(a) an input channel configured to receive a fluid;

(b) two or more levels of branched channels fluidically connected to said input channel, wherein said two or more levels of branched channels are below said input channel, and wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels; and

(c) a plurality of outputs fluidically connected to said two or more levels of branched channels, wherein said plurality of outputs are below the two or more levels of branched channels; wherein each binary split slopes downward from the upper channel directly above it at the same angle.

16. The device of embodiment 15, wherein said angle is about 100° to about 140°.

17. The device of embodiment 15, wherein said angle is about 115° to about 125°.

18. The device of embodiment 15, further comprising one or more vents comprising a sterile filter.

19. The device of embodiment 15, wherein an inner surface of said input channel or said two or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

20. The device of embodiment 19, wherein said raised structure comprises a herringbone pattern.

21. The device of embodiment 15, wherein said system is comprised of an optically transparent material. 22. The device of embodiment 15, wherein said fluid comprises solids.

23. The device of embodiment 22, wherein a concentration of solids in each output is within 15% of a mean concentration of solids of said plurality of outputs.

24. The device of embodiment 22, wherein a concentration of solids in each output is within 10% of a mean concentration of solids of said plurality of outputs.

25. The device of embodiment 22, wherein said solids comprise one or more cells.

26. The device of embodiment 15, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 10% of a mean volume of said plurality of outputs.

27. The device of embodiment 15, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 5% of a mean volume of said plurality of outputs.

28. A method for distributing a fluid, comprising:

(a) providing a vertical-flow fluid distribution device comprising an input channel, one or more levels of branched channels fluidically connected to said input channel;

(b) directing a fluid to said input channel, thereby allowing said fluid to flow in a substantially vertical direction from said input channel through said one or more levels of branched channels; and

(c) producing a plurality of output streams, wherein the volume of said fluid in each outlet stream is within 10% of a mean volume of said plurality of outlet streams.

29. The method of embodiment 28, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

30. The method of embodiment 28, wherein each binary split slopes downward from the upper channel directly above it at the same angle. 31. The method of embodiment 28, wherein said angle is about 100° to about 140°.

32. The method of embodiment 28, wherein said angle is about 115° to about 125°.

33. The method of embodiment 28, further comprising releasing air from said vertical-flow fluid distribution system through one or more vents, which consists of a sterile filter

34. The method of embodiment 28, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

35. The method of embodiment 34, wherein said raised structure comprises a herringbone pattern.

36. The method of embodiment 28, wherein said vertical-flow fluid distribution device is comprised of an optically transparent material.

37. The method of embodiment 28, wherein said fluid comprises solids.

38. The method of embodiment 37, wherein a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of said plurality of outlet streams.

39. The method of embodiment 37, wherein a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of said plurality of outlet streams.

40. The method of embodiment 37, wherein said solids comprise one or more cells.

41. The method of embodiment 36, further comprising optically observing a flow pattern of said fluid through said optically transparent material.

42. A vertical-flow fluid distribution system, comprising:

(a) a primary fluid distribution device configured to receive a fluid; and

(b) a plurality of secondary fluid distribution devices fluidically connected to said primary fluid distribution device, wherein said primary fluid distribution device is configured to distribute said fluid between each secondary fluid distribution device; wherein said vertical-flow distribution system is configured to output said fluid in a plurality of output streams, and wherein a volume of said fluid in each outlet stream is within 10% of a mean volume of said plurality of outlet streams.

43. The system of embodiment 42, wherein said primary fluid distribution device comprises: an input channel configured to receive a fluid and one or more levels of branched channels fluidically connected to said input channel, wherein said one or more levels of branched channels are below said input channel.

44. The system of embodiment 43, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

45. The system of embodiment 44, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

46. The system of embodiment 45, wherein said angle is about 100° to about 140°.

47. The system of embodiment 45, wherein said angle is about 115° to about 125°.

48. The system of embodiment 43, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

49. The system of embodiment 48, wherein said raised structure comprises a herringbone pattern.

50. The system of embodiment 42, wherein said primary fluid distribution device or said plurality of secondary fluid distribution devices are comprised of an optically transparent material.

51. The system of embodiment 42, wherein a secondary fluid distribution device of said plurality of secondary fluid distribution devices comprises an input channel configured to receive a fluid and one or more levels of branched channels fluidically connected to said input channel, wherein said one or more levels of branched channels are below said input channel. 52. The system of embodiment 51, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

53. The system of embodiment 52, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

54. The system of embodiment 53, wherein said angle is about 100° to about 140°.

55. The system of embodiment 53, wherein said angle is about 115° to about 125°.

56. The system of embodiment 51, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

57. The system of embodiment 56, wherein said raised structure comprises a herringbone pattern.

58. The system of embodiment 51, wherein said primary fluid distribution device distributes said fluid to a secondary fluid distribution device of said plurality of secondary fluid distribution devices via an input channel of said secondary fluid distribution device.

59. The system of embodiment 42, wherein said primary fluid distribution device is fluidically connected to said plurality of second fluid distribution devices through a plurality of tubes.

60. The system of embodiment 42, wherein a secondary fluid distribution device of said plurality of secondary fluid distribution devices outputs an outlet streams an outlet stream of said plurality of outlet streams into a container.

61. The system of embodiment 60, wherein when in said container, said output stream of said plurality of outlet streams is not fluidically connected with another outlet stream of said plurality of outlet streams. 62. The system of embodiment 60, wherein said container is fluidically connected to said secondary a secondary fluid distribution device of said plurality of secondary fluid distribution devices via tubing.

63. The system of embodiment 60, wherein said container comprises a centrifuge tube, cryobag, freezing bag, freeze-drying vial, or injection vial.

64. The system of embodiment 42, wherein said fluid comprises solids.

65. The system of embodiment 64, wherein a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of said plurality of outlet streams.

66. The system of embodiment 64, wherein a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of said plurality of outlet streams.

67. The system of embodiment 64, wherein said solids comprise one or more cells.

68. A method for distributing a fluid, comprising:

(a) providing a primary fluid distribution device and a plurality of secondary fluid distribution devices fluidically connected to said primary fluid distribution device, wherein said primary fluid distribution device is configured to distribute said fluid between each secondary fluid distribution device;

(b) directing a fluid to said primary fluid distribution device, thereby allowing said fluid to flow in a substantially vertical direction from said primary fluid distribution device through said plurality of secondary fluid distribution devices; and

(c) producing a plurality of output streams, wherein the volume of said fluid in each outlet stream is within 10% of a mean volume of said plurality of outlet stream

69. The method of embodiment 68, wherein said fluid flows from said primary fluid distribution device to said plurality of secondary fluid distribution devices through tubing. 70. The method of embodiment 68, further comprising releasing air from said primary fluid distribution device or said plurality of secondary fluid distribution devices through one or more vents.

71. The method of embodiment 68, wherein said primary fluid distribution device or said plurality of secondary fluid distribution is comprised of an optically transparent material.

72. The method of embodiment 71, further comprising optically observing a flow pattern of said fluid through said optically transparent material.

73. The method of embodiment 68, wherein said fluid comprises solids.

74. The method of embodiment 73, wherein a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of said plurality of outlet streams.

75. The method of embodiment 73, wherein a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of said plurality of outlet streams.

76. The method of embodiment 73, wherein said solids comprise one or more cells.

77. The method of embodiment 68, further comprising collecting an outlet stream of said plurality of outlet streams in a container, wherein when in said container, said output stream is not in fluid communication with another outlet stream of said plurality of outlet streams.

78. The method of embodiment 77, wherein said container comprises a well of a well-plate.

79. The method of embodiment 77, wherein said container comprises a test tube.

80. A non-transitory computer readable medium comprising machine-executable code that, upon execution by one or more computer processors, implements a method for distributing a fluid, the method comprising:

(a) directing a fluid to an input channel of a flow distribution device, wherein said flow distribution device comprises the input channel and a plurality of secondary channels, wherein each secondary channel of said plurality of secondary channels is fluidically connected to said input channel via a flow spliter;

(b) allowing said fluid to flow through said flow spliter and said plurality of second channels; and

81. The device of embodiment 1, wherein the number of outputs is an odd-number (e.g. 1 input to 3 outputs).

82. The device of embodiment 1, wherein a branched channel of the plurality of branched channels has a curved surface

83. The device of embodiment 1, wherein said input channel is located on a center axis of said device.

Claims

1. A vertical-flow fluid distribution device, comprising:

(a) an input channel configured to receive a fluid;

2. The device of claim 1, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

3. The device of claim 2, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

4. The device of claim 3, wherein said angle is about 100° to about 140°.

5. The device of claim 3 or claim 4, wherein said angle is about 115° to about 125°.

6. The device of any preceding claim, further comprising one or more vents comprising a sterile filter.

7. The device of any preceding claim, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

8. The device of claim 7, wherein said raised structure comprises a herringbone pattern.

9. The device of any preceding claim, wherein said system comprises an optically transparent material.

10. The device of any preceding claim, wherein said fluid comprises solids.

11. The device of claim 10, wherein a concentration of solids in each output is within 15% of a mean concentration of solids of said plurality of outputs.

12. The device of claim 10 or claim 11, wherein a concentration of solids in each output is within 10% of a mean concentration of solids of said plurality of outputs.

13. The device of any one of claims 10 to 12, wherein said solids comprise one or more cells.

14. The device of any preceding claim, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 5% of a mean volume of said plurality of outputs.

15. A vertical-flow fluid distribution device, comprising:

(a) an input channel configured to receive a fluid;

16. The device of claim 15, wherein said angle is about 100° to about 140°.

17. The device of claim 15 or claim 16, wherein said angle is about 115° to about 125°.

18. The device of any one of claims 15 to 17, further comprising one or more vents comprising a sterile filter.

19. The device of any one of claims 15 to 18, wherein an inner surface of said input channel or said two or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

20. The device of claim 19, wherein said raised structure comprises a herringbone pattern.

21. The device of any one of claims 15 to 20, wherein said system is comprised of an optically transparent material.

22. The device of any one of claims 15 to 21, wherein said fluid comprises solids.

23. The device of claim 22, wherein a concentration of solids in each output is within 15% of a mean concentration of solids of said plurality of outputs.

24. The device of claim 22 or claim 23, wherein a concentration of solids in each output is within 10% of a mean concentration of solids of said plurality of outputs.

25. The device of any one of claims 22 to 24, wherein said solids comprise one or more cells.

26. The device of any one of claims 15 to 25, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 10% of a mean volume of said plurality of outputs.

27. The device of any one of claims 15 to 26, wherein said device is configured to distribute a volume of the fluid through each output of the plurality of outputs, and wherein a volume of said fluid in each output is within 5% of a mean volume of said plurality of outputs.

28. A method for distributing a fluid, comprising:

(a) providing a vertical-flow fluid distribution device comprising an input channel, one or more levels of branched channels fluidically connected to said input channel; (b) directing a fluid to said input channel, thereby allowing said fluid to flow in a substantially vertical direction from said input channel through said one or more levels of branched channels; and

29. The method of claim 28, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

30. The method of claim 28, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

31. The method of claim 30, wherein said angle is about 100° to about 140°.

32. The method of claim 30 or claim 31, wherein said angle is about 115° to about 125°.

33. The method of any one of claims 28 to 32, further comprising releasing air from said vertical-flow fluid distribution system through one or more vents, which consists of a sterile filter

34. The method of any one of claims 28 to 33, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

35. The method of claim 34, wherein said raised structure comprises a herringbone pattern.

36. The method of any one of claims 28 to 35, wherein said vertical -flow fluid distribution device is comprised of an optically transparent material.

37. The method of any one of claims 28 to 26, wherein said fluid comprises solids.

38. The method of claim 37, wherein a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of said plurality of outlet streams.

39. The method of claim 37 or claim 38, wherein a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of said plurality of outlet streams.

40. The method of any one of claims 37 to 39, wherein said solids comprise one or more cells.

41. The method of any one of claims 36 to 40, further comprising optically observing a flow pattern of said fluid through said optically transparent material.

42. A vertical-flow fluid distribution system, comprising:

(a) a primary fluid distribution device configured to receive a fluid; and

43. The system of claim 42, wherein said primary fluid distribution device comprises: an input channel configured to receive a fluid and one or more levels of branched channels fluidically connected to said input channel, wherein said one or more levels of branched channels are below said input channel.

44. The system of claim 43, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

45. The system of claim 44, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

46. The system of claim 45, wherein said angle is about 100° to about 140°.

47. The system of claim 45 of claim 46, wherein said angle is about 115° to about 125°.

48. The system of any one of claims 43 to 47, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

49. The system of claim 48, wherein said raised structure comprises a herringbone pattern.

50. The system of any one of claims 42 to 49, wherein said primary fluid distribution device or said plurality of secondary fluid distribution devices are comprised of an optically transparent material.

51. The system of any one of claims 42 to 50, wherein a secondary fluid distribution device of said plurality of secondary fluid distribution devices comprises an input channel configured to receive a fluid and one or more levels of branched channels fluidically connected to said input channel, wherein said one or more levels of branched channels are below said input channel.

52. The system of claim 51, wherein each level of branched channels is separated from an upper channel by a binary split configured to distribute said fluid between two lower channels.

53. The system of claim 52, wherein each binary split slopes downward from the upper channel directly above it at the same angle.

54. The system of claim 53, wherein said angle is about 100° to about 140°.

55. The system of claim 53 or claim 54, wherein said angle is about 115° to about 125°.

56. The system of any one of claims 51 to 55, wherein an inner surface of said input channel or said one or more levels of branched channels comprises a raised structure configured to homogenize flow of said fluid through said input channel.

57. The system of claim 56, wherein said raised structure comprises a herringbone pattern.

58. The system of any one of claims 51 to 57, wherein said primary fluid distribution device distributes said fluid to a secondary fluid distribution device of said plurality of secondary fluid distribution devices via an input channel of said secondary fluid distribution device.

59. The system of any one of claims 42 to 58, wherein said primary fluid distribution device is fluidically connected to said plurality of second fluid distribution devices through a plurality of tubes.

60. The system of any one of claims 42 to 59, wherein a secondary fluid distribution device of said plurality of secondary fluid distribution devices outputs an outlet streams an outlet stream of said plurality of outlet streams into a container.

61. The system of claim 60, wherein when in said container, said output stream of said plurality of outlet streams is not fluidically connected with another outlet stream of said plurality of outlet streams.

62. The system of claim 60 or claim 61, wherein said container is fluidically connected to said secondary a secondary fluid distribution device of said plurality of secondary fluid distribution devices via tubing.

63. The system of any one of claims 60 to 62, wherein said container comprises a centrifuge tube, cryo-bag, freezing bag, freeze-drying vial, or injection vial.

64. The system of any one of claims 42 to 63, wherein said fluid comprises solids.

65. The system of claim 64, wherein a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of said plurality of outlet streams.

66. The system of claim 64 or claim 65, wherein a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of said plurality of outlet streams.

67. The system of any one of claims 64 to 66, wherein said solids comprise one or more cells.

68. A method for distributing a fluid, comprising:

(a) providing a primary fluid distribution device and a plurality of secondary fluid distribution devices fluidically connected to said primary fluid distribution device, wherein said primary fluid distribution device is configured to distribute said fluid between each secondary fluid distribution device; (b) directing a fluid to said primary fluid distribution device, thereby allowing said fluid to flow in a substantially vertical direction from said primary fluid distribution device through said plurality of secondary fluid distribution devices; and

69. The method of claim 68, wherein said fluid flows from said primary fluid distribution device to said plurality of secondary fluid distribution devices through tubing.

70. The method of claim 68 or claim 69, further comprising releasing air from said primary fluid distribution device or said plurality of secondary fluid distribution devices through one or more vents.

71. The method of any one of claims 68 to 70, wherein said primary fluid distribution device or said plurality of secondary fluid distribution is comprised of an optically transparent material.

72. The method of claim 71, further comprising optically observing a flow pattern of said fluid through said optically transparent material.

73. The method of any one of claims 68 to 72, wherein said fluid comprises solids.

74. The method of claim 73, wherein a concentration of solids in each outlet stream is within 15% of a mean concentration of solids of said plurality of outlet streams.

75. The method of claim 73 or claim 74, wherein a concentration of solids in each outlet stream is within 10% of a mean concentration of solids of said plurality of outlet streams.

76. The method of any one of claims 73 to 75, wherein said solids comprise one or more cells.

77. The method of any one of claims 68 to 76, further comprising collecting an outlet stream of said plurality of outlet streams in a container, wherein when in said container, said output stream is not in fluid communication with another outlet stream of said plurality of outlet streams.

78. The method of claim 77, wherein said container comprises a well of a well-plate.

79. The method of claim 77, wherein said container comprises a test tube.

(a) directing a fluid to an input channel of a flow distribution device, wherein said flow distribution device comprises the input channel and a plurality of secondary channels, wherein each secondary channel of said plurality of secondary channels is fluidically connected to said input channel via a flow splitter;

(b) allowing said fluid to flow through said flow splitter and said plurality of second channels; and

81. The device of any preceding claim, wherein the number of outputs is an odd-number (e.g. 1 input to 3 outputs).

82. The device of any preceding claim, wherein a branched channel of the plurality of branched channels has a curved surface.

83. The device of any preceding claim, wherein said input channel is located on a center axis of said device.