Systems and Methods for Automated Single Cell Cytological Classification in Flow

Patent metadata

Field	Value
Publication number	US-2017333902-A1
Application number	US-201715600618-A
Country	US
Kind code	A1
Filing date	May 19, 2017
Priority date	May 19, 2016
Publication date	Nov 23, 2017
Grant date	—

Abstract

Systems and methods in accordance with various embodiments of the invention are capable of rapid analysis and classification of cellular samples based on cytomorphological properties. In several embodiments, cells suspended in a fluid medium are passed through a microfluidic channel, where they are focused to a single stream line and imaged continuously. In a number of embodiments, the microfluidic channel establishes flow that enables individual cells to each be imaged at multiple angles in a short amount of time. A pattern recognition system can analyze the data captured from high-speed images of cells flowing through this system and classify target cells. In this way, the automated platform creates new possibilities for a wide range of research and clinical applications such as (but not limited to) point of care services.

First claim

What is claimed is:

1 . A cytological classification system comprising:
an imaging system; a flow cell comprising:
an inlet;
an outlet; and
a microfluidic channel comprising an imaging region, wherein the microfluidic channel receives flow via the inlet and having channel walls formed to:
focus cells from a sample into a single stream line;
space cells within a single stream line; and
rotate cells within a single stream line; and

a perfusion system configured to inject a sample into the flow cell via the inlet.

2 . The cytological classification system of claim 1 , wherein a sample injected into the flow cell includes a concentration of cells within the range of 1×10 5 cells/mL and 5×10 5 cells/mL.

3 . The cytological classification system of claim 1 , wherein a sample injected into the flow cell includes cells from the group consisting of fixed cells and cells stained with colored dyes.

4 . The cytological classification system of claim 1 , wherein the flow cell is implemented as a microfluidic device.

5 . The cytological classification system of claim 1 , wherein the imaging system comprises a light source configured to illuminate the imaging region of the microfluidic channel.

6 . The cytological classification system of claim 5 , wherein the imaging system further comprises an objective lens system configured to magnify the cells passing through the imaging region of the microfluidic channel.

7 . The cytological classification system of claim 5 , wherein the imaging system further comprises a high-speed camera system configured to capture images at between 100,000 and 500,000 frames/s.

8 . The cytological classification system of claim 1 , wherein the microfluidic channel is formed so that the imaging system captures a sequence of images of a rotating cell within the imaging region of the microfluidic channel that provides full 360° views of the cell.

9 . The cytological classification system of claim 1 , wherein the imaging system captures at least 10 images of a cell within the imaging region of the microfluidic channel.

10 . The cytological classification system of claim 1 , wherein the imaging system captures of images of at least 1000 cells/second and the computing system classifies at least 1000 cells/second.

11 . The cytological classification system of claim 1 , wherein the microfluidic channel further comprises a filtration region.

12 . The cytological classification system of claim 1 , wherein a subsection of the channel walls comprises a focusing region formed to focus cells from a sample into a single stream line of cells using inertial lift forces.

13 . The cytological classification system of claim 12 , wherein the inertial lift forces act on cells at Reynolds numbers where laminar flow occurs.

14 . The cytological classification system of claim 12 , wherein the focusing region includes contracted and expanded sections.

15 . The cytological classification system of claim 14 , wherein the contracted and expanded sections have an asymmetrical periodic structure.

16 . The cytological classification system of claim 1 , wherein a subsection of the channel walls comprises an ordering region formed to space cells within a single stream line using inertial lift forces and secondary flows that exert drag forces on the cells.

17 . The cytological classification system of claim 16 , wherein the ordering region forms at least one pinching region.

18 . The cytological classification system of claim 16 , wherein the ordering region forms a sequence of curved channels and pinching regions.

19 . The cytological classification system of claim 1 , wherein a subsection of the channel walls comprises a cell rotation region formed to rotate cells by applying a velocity gradient to the cells within the single stream line of cells.

20 . The cytological classification system of claim 19 , wherein the cell rotation region applies a velocity gradient to cells using a co-flow.

21 . The cytological classification system of claim 19 , wherein the cell rotation region applies a velocity gradient to cells by increasing at least one dimension of the channel.

22 . A cytological classification system comprising:
a two-layered flow cell comprising:
an inlet;
an outlet; and
a microfluidic channel comprising:
a focusing region for focusing cells from a sample into a single stream line;
an ordering region for spacing cells within a single stream line;
a cell rotation region for rotating cells within a single stream line; and
an imaging region that provides a field of view of rotating cells;

a perfusion system configured to inject a sample into the flow cell via the inlet; an imaging system comprising:
a camera configured to collect images of the imaging region;
a light source for illuminating the imaging region; and
an objective lens system configured to provide magnification of the imaging region; and
a computing system configured to receive images from the imaging system and to analyze the received images.

Assignees

• Univ Leland Stanford Junior

Inventors

• Masaeli Mahdokht
• Ashley Euan A

Classifications

• G06V20/698

  Matching; Classification · CPC title

• B01L3/50273

  characterised by the means or forces applied to move the fluids · CPC title

• B01L2400/086

  using baffles or other fixed flow obstructions · CPC title

• B01L2300/0681

  Filter · CPC title

• G01N15/1484

  microstructural devices · CPC title