Cellular diagnostic arrays, methods of using and processing for producing same

• US 7,208,120 B2
• Filed: 09/27/2001
• Issued: 04/24/2007
• Est. Priority Date: 09/27/2000
• Status: Expired due to Term

• Current Assignee: The Trustees of Princeton University (Princeton University), Trustees of Boston University
• Original Assignee: Trustees of Boston University
• Inventors: Yoshida, Tatsuro, Frank, Michael G., Derganc, Jure, Bitensky, Mark W., Gabel, Christopher V
• Primary Examiner(s): Strzelecka; Teresa E.
• Application Number:
  US09/965,644
• Publication Number:
  US 20020142285A1
• Time in Patent Office: 2,035 Days
• Field of Search: 435/4, 435/6, 435/7.24, 422/58, 422/68.1, 422/101, 422/55, 422/81, 422/82.08
• US Class Current: 422/68.1
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00657 One-dimensional arrays
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/014 Reticulocytes
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2015/1495 Deformation of particles
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

A cellular diagnostic array has been developed that provides accurate and reproducible measurements of the physical parameters of individual cells en masse, including cell membrane surface area, cell volume, and excess membrane. Three differing patterns are employed. In the first design, a gradient array having rectangular shaped channels is used to capture cells and readily show the ratio of volume to surface area for the captured cell. The second silicone rubber array provided by the present invention employs an array of wedge shaped pipettes, each of which captures an individual cell. The position of the cell within a wedge is enough to rapidly determine the cell'"'"'s actual surface area and volume. A third type of cellular diagnostic array according to the present invention employs an array of channels designed to deform a cell to study how well the cell responds to repeated deformations.

Claims

1. An array comprising a plurality of individual microchannels for capturing an individual cell therein, wherein each individual microchannel includes an entry portion for receiving said cell and an exit portion that said cell may pass through, wherein the plurality of microchannels are arranged in rows and columns and wherein there are at least two rows and at least two columns, wherein the exit portion of each individual microchannel faces the entry portion of the microchannel in a successive row.

2. The array of claim 1, wherein said microchannels each have a depth of 0.8 to 6.0 microns, a length of between 10 microns to 210 microns, said length including an entry portion for receiving said individual cell and an exit portion that said individual cell may pass through, said entry portion having an entry width of between 2.5 microns to 25 microns, and said exit portion having an exit width of between 0.5 microns to 7 microns.

3. The array of claim 1, wherein said microchannels are wedge-shaped, wherein the wedge shaped microchannels have a three dimensional entry portion that is wider than a three dimensional exit portion.

4. The array of claim 2, wherein said length is about 60 microns, said entry width is about 3.7 microns, and said exit width is about 1.5 microns and depth of about 3.4 microns.

5. The array of claim 2, wherein said length is about 35 microns, said entry width is about 3.6 microns, and said exit width is about 1.4 microns.

6. The array of claim 2, wherein said length is about 100 microns, said entry width is about 4.5 microns, and said exit width is about 1.5 microns.

7. The array of claim 2, wherein said length is about 16 microns, said entry width is about 3.6 microns, and said exit width is about 1.4 microns.

8. The array of claim 1, further comprising shunt channels, wherein said shunt channels comprise individual microchannels arranged in at least one row and at least two columns, wherein said microchannels are adapted to allow said cell to enter the entry portion and exit the exit portion in order to bypass an individual microchannel occupied with a cell.

9. The array of claim 8, wherein said shunt channels have a length of between 10 microns and 100 microns.

10. The array of claim 1, wherein said microchannels have cross-sectional dimensions adapted to temporarily deform a cell passing therethrough.

11. The array of claim 1, further comprising a means for moving said cells through said microchannels.

12. The array of claim 11, wherein said means for moving said cells through said microchannels comprises a vacuum pump for pulling said cell through said microchannels, negative pressure generated by connected water columns, or a peristaltic pump for driving said cell through the channels.

14. The array of claim 3, wherein the dimensions of the wedge-shaped microchannel are defined by area and volume, wherein the area and volume of a cell are known and the length, width and height of the microchannel is varied so that a target blood cell may enter, but not exit.

13. A gradient army for analyzing a plurality of cells comprising at least three rows of microchannels and at least two columns of microchannels, wherein the microchannels have an entry portion for receiving cells and an exit portion whereby cells may pass through, wherein the exit portion of the microchannels in the first row face the entry portion of the microchannels in a second row and the exit portion of the microchannels in the second row face the entry portion of the microchannel in a third row, and wherein the width of the microchannels in the first row is wider than the cells to be analyzed such that cells enter the entry portion and exit the exit portion of the microchannels in the first row and enter the entry portion of the microchannels in the second row, and wherein the microchannels in the first row have a first width which is larger compared to the width of the microchannels in the second row, wherein cells are either trapped within a microchannel in the second row or pass through the exit portion of the microchannel in the second row and enter the entry portion of them microchannel in the third row, wherein cells that pass through the exit portion of the second row enter the entry portion of the microchannels in the third row.

15. An array comprising a plurality of individual microchannels for capturing an individual cell therein, wherein each individual microchannel includes an entry portion for receiving said cell and an exit portion that said cell may pass through, wherein the plurality of microchannels are arranged in rows and columns and wherein there are at least two rows and at least two columns, wherein the exit portion of each individual microchannel faces the entry portion of the microchannel in a successive row, wherein each individual microchannel is a wedge shape, wherein the wedge shaped microchannels have a three dimensional entry portion that is wider than a three dimensional exit portion.