IP Library Granted Patent US 9,752,181
Granted Patent B2
US 9,752,181 · App. 14/163,185 · Granted Sep 5, 2017

System and method for capturing and analyzing cells

Inventors: Kalyan Handique (Ann Arbor, MI); Priyadarshini Gogoi (Ann Arbor, MI); Saedeh Sepehri Javdani (Ypsilanti, MI); Yi Zhou (Plymouth, MI)
Assignee: DeNovo Sciences, Inc.
C12Q1/6841C12Q1/686C12Q1/6844G01N33/48721G01N33/574G01N33/57492G01N27/44782G01N2333/70585G01N2333/70596
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Quick Facts
Patent No.
US 9,752,181
App. No.
14/163,185
Granted
Sep 5, 2017
Kind
B2
Abstract

A system and method for capturing and analyzing a set of cells, comprising: an array including a set of parallel pores, each pore including a chamber including a chamber inlet and a chamber outlet, and configured to hold a single cell, and a pore channel fluidly connected to the chamber outlet; an inlet channel fluidly connected to each chamber inlet of the set of parallel pores; an outlet channel fluidly connected to each pore channel of the set of parallel pores; a set of electrophoresis channels fluidly coupled to the outlet channel, configured to receive a sieving matrix for electrophoretic separation; and a set of electrodes including a first electrode and a second electrode, wherein the set of electrodes is configured to provide an electric field that facilitates electrophoretic analysis of the set of cells.

Claims (18)

1. A method for capturing and analyzing a set of cells, comprising:

capturing the set of cells, including a subpopulation of cells, at a set of parallel pores of a substrate, each pore in the set of parallel pores including a chamber comprising:

an inlet and an outlet, and a set of walls defining a chamber volume directly corresponding to the volume of a single cell of the set of cells and thereby configured to hold the single cell of the set of cells;

transmitting a reagent volume to the set of parallel pores at an inlet channel fluidly coupled to the set of parallel pores, wherein the reagent volume is configured to distinguish the subpopulation of cells from the set of cells;

transmitting excitation wavelengths of light into each pore of the set of parallel pores, thereby enabling identification of at least one cell of the subpopulation of cells;

guiding a cell removal tool into the inlet channel and to a pore of the set of parallel pores containing a cell of the subpopulation of cells;

transmitting a force configured to drive the cell of the subpopulation of cells from the pore and into the cell removal tool; and

performing reverse transcription polymerase chain reaction (RT-PCR) for the cell of the subpopulation of cells, wherein the cell comprises a circulating stem cell (CSC), and wherein performing RT-PCR includes determining threshold cycle values for a group of target genes selected from a group consisting of Her2, ALDH1, and TWIST1.

2. The method of claim 1 , wherein capturing the set of cells includes capturing a set of circulating tumor cells (CTCs), and wherein the subpopulation of cells includes a subpopulation of circulating stem cells (CSCs).

3. The method of claim 2 , wherein transmitting the reagent volume includes transmitting an antibody cocktail including CD24 and CD44 antibodies.

4. The method of claim 3 , wherein transmitting the antibody cocktail further includes transmitting an additional antibody to the set of parallel pores, wherein the additional antibody enables identification of contaminating cells captured at the set of parallel pores.

5. The method of claim 3 , further including facilitating antigen retrieval of the set of cells, with at least one of citrate buffer, sodium dodecyl sulfate, and enzymatic treatment.

6. The method of claim 3 , further including transmitting a bright field stain with the antibody cocktail, thereby mitigating interference due to spectral overlapping of fluorophores.

7. The method of claim 3 , wherein transmitting excitation wavelengths of light further includes enabling identification of a CD44 + /CD24 − phenotype characterizing cells of the subpopulation of CSCs.

8. The method of claim 1 , wherein transmitting excitation wavelengths of light includes transmitting light by way of a reflector configured to reflect incident light at a substantially 90 degree angle longitudinally into each pore of the set of parallel pores.

9. The method of claim 1 , wherein guiding the cell removal tool includes receiving the cell removal tool into the inlet channel at a self-sealing portion configured to form a hermetic seal about the cell removal tool upon penetration of the inlet channel by the cell removal tool.

10. The method of claim 1 , wherein transmitting the force includes providing a positive pressure at an outlet channel fluidly coupled to the set of parallel pores, thereby pushing the cell of the subpopulation of cells into the cell removal tool.

11. The method of claim 1 , wherein transmitting the force includes providing a negative pressure at the cell removal tool, thereby drawing the cell of the subpopulation of cells into the cell removal tool.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 5, 2020
From: CELSEE, INC.
To: BIO-RAD LABORATORIES, INC.
Reel/Frame 054269/0742 →
CHANGE OF NAME Recorded Sep 24, 2020
From: CELSEE DIAGNOSTICS, INC.
To: CELSEE, INC.
Reel/Frame 053881/0555 →
CHANGE OF NAME Recorded Aug 27, 2018
From: DENOVO SCIENCES, INC.
To: CELSEE DIAGNOSTICS, INC.
Reel/Frame 047470/0615 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 3, 2014
From: HANDIQUE, KALYAN; GOGOI, PRIYADARSHINI; ZHOU, YI; JAVDANI, SAEDEH SEPEHRI
To: DENOVO SCIENCES, INC.
Reel/Frame 032120/0512 →
Continuity (3)
Provisional Application 61757141 · Jan 26, 2013
Provisional Application 61757139 · Jan 26, 2013
Related Publication 20140212881A1 · Jul 31, 2014