公开(公告)号：US20190119737A1

公开(公告)日：2019-04-25

申请号：US16301687

申请日：2017-05-16

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Janay Kong ,  Aydogan Ozcan ,  Omai Garner

IPC: C12Q1/6844 ,  C09B29/16 ,  C09B23/04 ,  C09B15/00 ,  C09B69/04

Abstract: A fluorescent dye and quencher mixture for reporting on nucleic acid amplification from a sample includes a fluorescent intercalating dye, a dye sequestering or quenching agent such as hydroxynapthol blue (HNB) or caffeine, and primers, dNTPs, and a nucleic acid polymerizing enzyme or fragment thereof. The presence of the dye in combination with the dye sequestering or quenching agent improves the overall dynamic range of the fluorescent signal as well as shortens the time needed for visualization or image capture of amplified nucleic acid. The fluorescent dye and quencher mixture also enables the detection of nucleic acids in samples having low copy numbers.

公开(公告)号：US20180266452A1

公开(公告)日：2018-09-20

申请号：US15763765

申请日：2016-09-30

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Chueh-Yu Wu

IPC: F15D1/14 ,  B01J19/00 ,  B01L3/00 ,  B01J19/12 ,  G03F7/027

CPC classification number: F15D1/14 ,  B01J19/0093 ,  B01J19/123 ,  B01L3/502707 ,  B01L3/502761 ,  B01L3/502776 ,  B01L2200/12 ,  B01L2400/043 ,  B01L2400/086 ,  G03F7/027

Abstract: A method of forming three-dimensional shaped microparticles in a microfluidic device includes flowing a mixture of a monomer and photoinitiator in a microfluidic channel having a plurality of pillars disposed therein to define a flow stream having a pre-defined shape and temporarily stopping the same. One or more portions of the flow stream are polymerized by passing polymerizing light through one or more masks and onto the flow stream, the polymerization process forming a plurality of three-dimensional shaped microparticles. The three-dimensional shape of the microparticle may be geometrically complex by using non-rectangular 2D orthogonal shapes for the flow and/or masked light source. The microparticles may include protected regions on which cells can be adhered to and protected from shear forces. The flow stream is restarted to flush out the newly formed microparticles and prepare the device for the next cycle of particle formation.

公开(公告)号：US20160231224A1

公开(公告)日：2016-08-11

申请号：US14552256

申请日：2014-11-24

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Daniel R. Gossett ,  Henry T.K. Tse

IPC: G01N15/14 ,  G01N33/487 ,  B01L3/00

CPC classification number: G01N15/1463 ,  B01L3/502715 ,  B01L3/50273 ,  B01L3/502776 ,  B01L2200/0605 ,  B01L2200/0647 ,  B01L2200/0652 ,  B01L2300/0654 ,  B01L2300/0864 ,  C12Q1/02 ,  G01N15/1404 ,  G01N15/1459 ,  G01N15/1484 ,  G01N33/487 ,  G01N33/574 ,  G01N2015/0065 ,  G01N2015/1006 ,  G01N2015/1415 ,  G01N2015/149 ,  G01N2015/1495 ,  G01N2015/1497 ,  G01N2800/7028 ,  G06F19/10

Abstract: A system is disclosed that enables the automated measurement of cellular mechanical parameters at high throughputs. The microfluidic device uses intersecting flows to create an extensional flow region where the cells undergo controlled stretching. Cells are focused into streamlines prior to entering the extensional flow region. In the extensional region, each cell's deformation is measured with an imaging device. Automated image analysis extracts a range of independent biomechanical parameters from the images. These may include cell size, deformability, and circularity. The single cell data that is obtained may then be used to in a variety of ways. Scatter density plots of deformability and circularity may be developed and displayed for the user. Mechanical parameters such as deformability and circularity may be gated or thresholded to identify certain cells of interest or sub-populations of interest. Similarly, the mechanical data obtained using the device may be used as cell signatures.

Abstract translation: 公开了一种能够以高产量自动测量细胞机械参数的系统。 微流体装置使用相交流来产生细胞经受受控拉伸的拉伸流动区域。 在进入延伸流动区域之前，细胞被聚焦成流线。 在拉伸区域中，每个单元的变形用成像装置测量。 自动图像分析从图像中提取一系列独立的生物力学参数。 这些可能包括细胞大小，变形能力和圆形度。 然后可以以各种方式使用所获得的单细胞数据。 可以为用户开发和显示可变形性和圆形度的散射密度图。 诸如可变形性和圆形度的机械参数可以被门控或阈值化以识别感兴趣的某些细胞或感兴趣的亚群。 类似地，使用该设备获得的机械数据可以用作单元签名。

公开(公告)号：US09333510B2

公开(公告)日：2016-05-10

申请号：US14346290

申请日：2012-09-27

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Mahdokht Masaeli ,  Elodie Sollier

IPC: B03B5/48 ,  G01N1/40 ,  G01N35/08 ,  G01N15/02 ,  B01L3/00 ,  G01N15/00

CPC classification number: B03B5/48 ,  B01L3/502746 ,  B01L3/502761 ,  B01L2200/0636 ,  B01L2200/0652 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2300/088 ,  B01L2400/0487 ,  G01N1/4077 ,  G01N15/02 ,  G01N35/08 ,  G01N2015/0065 ,  G01N2015/0288 ,  G01N2015/0294

Abstract: A particle sorting system includes an inlet and an inertial focusing microchannel disposed in a substrate and having a downstream expanding region at a distal end, wherein the inlet is connected to an upstream end of the microchannel. A source of different shaped particles is connected to the inlet, wherein the source of different shaped particles are configured for continuous introduction into the inlet. A plurality of outlets is connected to the microchannel at the downstream expanding region. Fluidic resistors are located in the respective outlets. Different resistances may be used in the outlets to capture enriched fractions of particles having particular particle shape(s).

Abstract translation: 颗粒分选系统包括设置在基板中的入口和惯性聚焦微通道，并且在远端具有下游扩张区域，其中入口连接到微通道的上游端。 不同成形颗粒的源连接到入口，其中不同成形颗粒的源被构造成用于连续引入入口。 多个出口在下游扩展区域连接到微通道。 流体电阻位于相应的出口。 可以在出口中使用不同的电阻来捕获具有特定颗粒形状的颗粒的富集部分。

公开(公告)号：US20140315287A1

公开(公告)日：2014-10-23

申请号：US14058028

申请日：2013-10-18

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Daniel R. Gossett ,  Henry T.K. Tse ,  Aram Chung

IPC: G01N15/14

CPC classification number: G01N15/1404 ,  G01N15/1434 ,  G01N15/1436 ,  G01N15/1459 ,  G01N15/147 ,  G01N15/1484 ,  G01N21/64 ,  G01N21/6428 ,  G01N21/645 ,  G01N33/50 ,  G01N33/5091 ,  G01N2015/0065 ,  G01N2015/1006 ,  G01N2015/1415 ,  G01N2015/1495 ,  G01N2021/6439 ,  G06K9/4604 ,  G06T7/0004 ,  G06T7/0016

Abstract: A system for deforming and analyzing a plurality of particles carried in a sample volume includes a substrate defining an inlet, configured to receive the sample volume, and an outlet; and a fluidic pathway fluidly coupled to the inlet and the outlet. The fluidic pathway includes a delivery region configured to receive the plurality of particles from the inlet and focus the plurality of particles from a random distribution to a focused state, a deformation region defining an intersection located downstream of the delivery region and coupled to the outlet, and wherein the deformation region is configured to receive the plurality of particles from the delivery region and to transmit each particle in the plurality of particles into the intersection from a single direction, a first branch fluidly coupled to the deformation region and configured to transmit a first flow into the intersection, and a second branch fluidly coupled to the deformation region and configured to transmit a second flow, substantially opposing the first flow, into the intersection, wherein the first flow and the second flow are configured to induce extension of one or more particles in the plurality of particles.

Abstract translation: 用于变形和分析载体在样品体积中的多个颗粒的系统包括限定入口的衬底，其构造成接收样品体积和出口; 以及流体连接到入口和出口的流体通路。 流体通道包括配置成从入口接收多个颗粒并将多个颗粒从随机分布聚焦到聚焦状态的输送区域，限定位于输送区域下游并且耦合到出口的交叉的变形区域， 并且其中所述变形区域被配置为从所述递送区域接收所述多个颗粒并且将多个颗粒中的每个颗粒从单个方向传播到所述交叉点，所述第一分支流体耦合到所述变形区域并且被配置为传输第一 流入所述十字路口的第二分支和流体耦合到所述变形区域并被配置为将基本上与所述第一流相对的第二流传送到所述交叉点的第二分支，其中所述第一流和所述第二流被配置为诱导一个或多个 多个颗粒中的颗粒。

公开(公告)号：US20140113324A1

公开(公告)日：2014-04-24

申请号：US14057942

申请日：2013-10-18

Applicant: The Regents Of The University Of California

Inventor： Dino Di Carlo ,  Daniel R. Gossett ,  Henry T.K. Tse ,  Aram Chung

IPC: G01N33/50

CPC classification number: G01N15/1404 ,  G01N15/1434 ,  G01N15/1436 ,  G01N15/1459 ,  G01N15/147 ,  G01N15/1484 ,  G01N21/64 ,  G01N21/6428 ,  G01N21/645 ,  G01N33/50 ,  G01N33/5091 ,  G01N2015/0065 ,  G01N2015/1006 ,  G01N2015/1415 ,  G01N2015/1495 ,  G01N2021/6439 ,  G06K9/4604 ,  G06T7/0004 ,  G06T7/0016

Abstract: A system for deforming and analyzing particles includes a substrate defining an inlet, and an outlet; a fluidic pathway fluidly coupled to the inlet and the outlet and defining a delivery region upstream of a deformation region configured to deform particles, wherein the fluidic pathway comprises a first branch configured to generate a first flow, and a second branch configured to generate a second flow that opposes the first flow, wherein an intersection of the first flow and the second flow defines the deformation region; a detection module including a sensor configured to generate a morphology dataset characterizing deformation of the particles, and a photodetector configured to generate a fluorescence dataset characterizing fluorescence of the particles; and a processor configured to output an analysis of the plurality of particles based at least in part on the deformation dataset and the fluorescent dataset for the plurality of particles.

Abstract translation: 用于变形和分析颗粒的系统包括限定入口的基板和出口; 流体路径，流体地联接到入口和出口，并且限定被配置为变形颗粒的变形区域上游的输送区域，其中流体路径包括被配置为产生第一流量的第一分支和被配置为产生第二流量的第二分支 所述第一流与所述第一流相对，其中所述第一流和所述第二流的交点限定所述变形区; 检测模块，其包括被配置为生成表征所述粒子的变形的形态学数据集的传感器，以及被配置为产生表征所述粒子的荧光的荧光数据集的光电检测器; 以及处理器，被配置为至少部分地基于所述多个粒子的所述变形数据集和所述荧光数据集来输出所述多个粒子的分析。

公开(公告)号：US20250155354A1

公开(公告)日：2025-05-15

申请号：US19025868

申请日：2025-01-16

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Daniel R. Gossett ,  Henry T.K. Tse

IPC: G01N15/1433 ,  B01L3/00 ,  C12Q1/02 ,  G01N15/01 ,  G01N15/10 ,  G01N15/14 ,  G01N15/1404 ,  G01N15/149 ,  G01N33/487 ,  G01N33/574 ,  G16B99/00

Abstract: A system is disclosed that enables the automated measurement of cellular mechanical parameters at high throughputs. The microfluidic device uses intersecting flows to create an extensional flow region where the cells undergo controlled stretching. Cells are focused into streamlines prior to entering the extensional flow region. In the extensional region, each cell's deformation is measured with an imaging device. Automated image analysis extracts a range of independent biomechanical parameters from the images. These may include cell size, deformability, and circularity. The single cell data that is obtained may then be used to in a variety of ways. Scatter density plots of deformability and circularity may be developed and displayed for the user. Mechanical parameters such as deformability and circularity may be gated or thresholded to identify certain cells of interest or sub-populations of interest. Similarly, the mechanical data obtained using the device may be used as cell signatures.

公开(公告)号：US12007321B2

公开(公告)日：2024-06-11

申请号：US17200728

申请日：2021-03-12

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Dino Di Carlo ,  Daniel R. Gossett ,  Henry T. K. Tse ,  Aram Chung

IPC: G01N15/14 ,  G01N15/1404 ,  G01N15/1434 ,  G01N21/64 ,  G01N33/50 ,  G06T7/00 ,  G01N15/01 ,  G01N15/10

CPC classification number: G01N15/147 ,  G01N15/1404 ,  G01N15/1434 ,  G01N15/1436 ,  G01N15/1459 ,  G01N15/1484 ,  G01N21/6428 ,  G01N21/645 ,  G01N33/50 ,  G01N33/5091 ,  G06T7/0004 ,  G01N15/01 ,  G01N2015/1006 ,  G01N2015/1415 ,  G01N2015/1495 ,  G01N21/64 ,  G01N2021/6439 ,  G06T7/0016

Abstract: A system for deforming and analyzing a plurality of particles carried in a sample volume includes a substrate defining an inlet, configured to receive the sample volume, and an outlet; and a fluidic pathway fluidly coupled to the inlet and the outlet. The fluidic pathway includes a delivery region configured to receive the plurality of particles from the inlet and focus the plurality of particles from a random distribution to a focused state, a deformation region defining an intersection located downstream of the delivery region and coupled to the outlet, and wherein the deformation region is configured to receive the plurality of particles from the delivery region and to transmit each particle in the plurality of particles into the intersection from a single direction, a first branch fluidly coupled to the deformation region and configured to transmit a first flow into the intersection, and a second branch fluidly coupled to the deformation region and configured to transmit a second flow, substantially opposing the first flow, into the intersection, wherein the first flow and the second flow are configured to induce extension of one or more particles in the plurality of particles.

公开(公告)号：US20230190995A1

公开(公告)日：2023-06-22

申请号：US17935096

申请日：2022-09-24

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Donald R. Griffin ,  Westbrook Weaver ,  Tatiana Segura ,  Dino Di Carlo ,  Philip Scumpia

IPC: A61L26/00 ,  A61L27/18 ,  A61L27/58 ,  A61K47/62 ,  A61K9/06 ,  A61K31/795 ,  A61L27/22 ,  A61L27/52 ,  A61L27/54 ,  A61L27/56

CPC classification number: A61L26/008 ,  A61L26/0019 ,  A61L26/0085 ,  A61L26/009 ,  A61L27/18 ,  A61L27/58 ,  A61K47/62 ,  A61K9/06 ,  A61K31/795 ,  A61L26/0066 ,  A61L26/0047 ,  A61L27/227 ,  A61L27/52 ,  A61L27/54 ,  A61L27/56 ,  A61L2430/34 ,  A61L2300/252 ,  A61L2300/412 ,  A61L2400/06 ,  A61L2430/00

Abstract: A microporous gel system for certain applications, including biomedical applications, includes an aqueous solution containing plurality of microgel particles including a biodegradable crosslinker. In some aspects, the microgel particles act as gel building blocks that anneal to one another to form a covalently-stabilized scaffold of microgel particles having interstitial spaces therein. In certain aspects, annealing of the microgel particles occurs after exposure to an annealing agent that is endogenously present or exogenously added. In some embodiments, annealing of the microgel particles requires the presence of an initiator such as exposure to light. In particular embodiments, the chemical and physical properties of the gel building blocks can be controlled to allow downstream control of the resulting assembled scaffold. In one or more embodiments, cells are able to quickly infiltrate the interstitial spaces of the assembled scaffold.

公开(公告)号：US20230173449A1

公开(公告)日：2023-06-08

申请号：US17996927

申请日：2021-04-26

Applicant: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Inventor： Joseph De Rutte ,  Dino Di Carlo ,  Sohyung Lee

IPC: B01J13/14 ,  G01N33/58

CPC classification number: B01J13/14 ,  G01N33/585

Abstract: A method of fabricating shaped particles is disclosed. The method involves generating a plurality of droplets within dispersion media (e.g., oil and surfactant), the plurality of droplets formed from a mixture of precursor materials that are in a miscible state. A stimulus or change of conditions is then introduced to the droplets so as to cause the mixture of precursor materials to become immiscible and phase-separate from one another. The phase-separated droplets are then crosslinked to form shaped particles. The stimulus or change of conditions may include one or more of the following: a change in temperature, a change in pH, a change in osmolarity, a change composition of the droplets, a change in the composition of the dispersion media. The shaped particles may be washed to remove un-crosslinked material and one or more affinity capture agents may be immobilized onto the shaped particles.