IP Library Granted Patent US 9,709,479
Granted Patent B2
US 9,709,479 · App. 14/523,376 · Granted Jul 18, 2017

Method and apparatus for tracking cell identity

Inventors: Marc Castellarnau (Charlestown, MA); Gregory Lee Szeto (Cambridge, MA); Darrell J. Irvine (Arlington, MA); John Christopher Love (Tbilisi, MA); Joel Voldman (Belmont, MA)
Assignee: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
G01N15/1463G01N15/10G01N35/00732G01N15/1484G01N2015/008G01N2015/0065G01N2015/1006G01N2015/149G01N2035/00811G01N2035/00831
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Quick Facts
Patent No.
US 9,709,479
App. No.
14/523,376
Granted
Jul 18, 2017
Kind
B2
Abstract

A method of tracking cell identity across analytical platforms uses stochastic barcoding (SB). SB uses a randomly generated code based on one or more of the number, color and position of beads encapsulated together with a set of cells of interest. SB use is demonstrated in an application where cells are transferred from a microwell array into a microtiter plate while keeping their identity, and obtained an average identification accuracy of 96% for transfer of 100 blocks. Model scaling of the method up to 1000 blocks demonstrated that SB is able to achieve approximately 90% accuracy.

Claims (28)

1. A method of tracking cell identity comprising:

adding a polymer solution with a plurality of markers into a block of a single cell microwell arrays;

encapsulating the single cell in a matrix including a plurality of markers; and

imaging the encapsulated cell to create a stochastic barcode determined by number, color, and position of the markers within the matrix, wherein the stochastic barcode is assigned to the single cell.

2. The method of claim 1 , wherein the markers include polymer beads.

3. The method of claim 2 , wherein the polymer beads include emissive polymer beads.

4. The method of claim 2 , wherein the polymer beads are colored.

5. The method of claim 4 , wherein the number of colors is two to six.

6. The method of claim 4 , wherein the number of colors is three.

7. The method of claim 4 , wherein the number of colors is four.

8. The method of claim 1 , wherein the matrix is a hydrogel.

9. The method of claim 1 , further comprising:

imaging of arrays to assign a random code based on the plurality of markers;

transferring arrays into a microtiter plate; and

imaging arrays after transfer to read the code.

10. The method of claim 9 , wherein the random code is assigned based on the number, color, size, and/or position of markers in the matrix.

11. The method of claim 9 , wherein the encapsulation step includes photopolymerizing or chemical polymerization or thermal polymerization.

12. The method of claim 9 , wherein assigning a random code includes using a machine learning algorithm.

13. The method of claim 9 , wherein the markers include polymer beads.

14. The method of claim 13 , wherein the polymer beads include emissive polymer beads.

15. The method of claim 14 , wherein the polymer beads are colored.

16. The method of claim 15 , wherein the number of colors is two to six.

17. The method of claim 15 , wherein the number of colors is three.

18. The method of claim 15 , wherein the number of colors is four.

19. The method of claim 9 , wherein the matrix is a hydrogel.

20. The method of claim 13 , wherein approximately one hundred cells in an array may be uniquely coded with an average of eight beads per encapsulated cell.

21. The method of claim 20 , wherein an average of fifteen beads per encapsulated cell yields an approximate 100 percent matching accuracy.

22. The method of claim 9 , wherein an average of fifteen beads per encapsulated cell yields an approximate 90 percent matching accuracy for one thousand cells.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 9, 2017
From: CASTELLARNAU, MARC; SZETO, GREGORY LEE; IRVINE, DARRELL J.; LOVE, JOHN CHRISTOPHER; VOLDMAN, JOEL
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 042663/0491 →
Continuity (2)
Provisional Application 61895732 · Oct 25, 2013
Related Publication 20150253237A1 · Sep 10, 2015