Chamber for imaging cells and method of use thereof

Patent metadata

Field	Value
Publication number	US-10479972-B2
Application number	US-201615567132-A
Country	US
Kind code	B2
Filing date	Apr 6, 2016
Priority date	Apr 20, 2015
Publication date	Nov 19, 2019
Grant date	Nov 19, 2019

Abstract

One aspect of the invention provides a sample chamber for use in the dynamic imaging of cells. In one embodiment, the chamber includes a holding mechanism for positioning the cells against a viewing window in the chamber. A housing is positioned against the viewing window to define the closed chamber. The housing may include access ports allowing for the perfusion of fluid through the chamber, the control of the internal temperature of the chamber and the micromanipulation and electrical stimulation of the cells.

First claim

We claim:

1. An apparatus for imaging cells comprising:
a chamber base assembly comprising a viewing window;
a cell-holding bay positioned on top of the viewing window, wherein the viewing window forms at least a portion of a base of the cell-holding bay;
a first cell-holding assembly extending through a first conduit in the chamber base assembly and having a first end positionable at a first position in the cell-holding bay;

a second cell-holding assembly extending through a second conduit in the chamber base assembly and having a first end positionable at a second position in the sample holding bay;
a chamber perfusion and manipulation manifold mounted on the chamber base assembly on top of the cell-holding bay,
wherein the first ends of the first and second cell-holding assemblies are movable between a first position holding the cells against the viewing window and a second position allowing placement and removal of the cells from the cell-holding bay.

2. The apparatus of claim 1 , wherein the first cell-holding assembly comprises:
a cell-holding wire forming the first end of the first cell-holding assembly;
a linkage member having a first end connecting to the cell-holding wire and extending through the first conduit to a second end at an exterior of the chamber base assembly: and
a handle connecting to the second end of the linkage member.

3. The apparatus of claim 2 , wherein the linkage member comprises:
a flexible member having a first end attaching to the handle; and
a cylindrical member positioned within the first conduit and having a first end attaching to a second end of the flexible member.

4. The apparatus of claim 3 , wherein the cell-holding wire is a step shaped wire comprising a first end portion, second end portion and a central portion joining the first end portion to the second end portion.

5. The apparatus of claim 4 , wherein first end portion of the cell-holding wire extends substantially perpendicularly from an eccentric hole in a second end of the cylindrical member and wherein rotation of the cylindrical member allows for positioning of the second end portion of the cell-holding wire to holding the cells against the viewing window or positioning of the second end portion of the cell-holding wire away from the viewing window.

6. The apparatus of claim 1 , further comprising a fluid entry line extending from the exterior of the chamber housing, through the chamber housing to a fluid entry line end positioned to deliver fluid into the cell-holding bay, and a fluid exit line extending from a fluid exit port in the cell-holding bay, through the chamber housing to a fluid aspiration port position exterior to the chamber housing.

7. The apparatus of claim 1 , further comprising a temperature control unit positioned in the chamber base and providing for temperature control of the cell-holding bay.

8. The apparatus of claim 1 , further comprising a first and a second electrical connector positioned on the base of the cell-holding bay, wherein the first and second electrical connectors are in electrical contact with electrical terminals exterior to the chamber and allow for electrical stimulation of a cell positioned in the cell-holding bay.

9. The apparatus of claim 1 , wherein the cell-holding bay comprises a depressed portion of the floor of the chamber base assembly and wherein a transparent window form at least a portion of the floor of the cell-holding bay.

10. The apparatus of claim 1 , wherein the chamber housing comprises a removable lid.

11. The apparatus of claim 1 , wherein at least a portion of at least one of the chamber base assembly and the chamber housing is made using 3D printing.

12. A microscope system for imaging cells comprising:
an optical microscope unit; and
a chamber comprising:
a chamber base assembly comprising a viewing window;
a cell-holding bay positioned on top of the viewing window;
a first cell-holding assembly extending through a first conduit in the chamber base assembly and having a first end positionable at a first position in the cell-holding bay;

a second cell-holding assembly extending through a second conduit in the chamber base assembly and having a first end positionable at a second position in the sample holding bay;
a chamber housing mounted on the chamber base assembly and enclosing the cell-holding bay,
wherein the first ends of the first and second cell-holding assemblies are movable between a first position holding the cells against the viewing window and a second position allowing removal of the cells from the cell-holding bay,
wherein the chamber is positioned to allow cells held against the viewing window to be viewed through the microscope unit.

13. A method of viewing a cell, the method comprising:
providing an apparatus comprising:
a chamber base assembly comprising a viewing window;
a cell-holding bay positioned on top of the viewing window, wherein the viewing window form a base of the cell-holding bay;
a first cell-holding assembly extending through a first conduit in the chamber base assembly and having a first end positionable at a first position in the cell-holding bay;
a second cell-holding assembly extending through a second conduit in the chamber base assembly and having a first end positionable at a second position in the sample holding bay;
a chamber housing mounted on the chamber base assembly and enclosing the cell-holding bay,
wherein the first ends of the first and second cell-holding assemblies are movable between a first position holding the cell against the viewing window and a second position allowing removal of the cell from the cell-holding bay;

placing the first ends of the first and second cell-holding assemblies in the second position;
positioning the cell on the base of the cell-holding bay between the viewing window and the first ends of the first and second cell-holding assemblies;
moving the first ends of the first and second cell-holding assemblies to the first position to contact differing portions of the cell so as to position the cell against the viewing window; and
viewing the cell through the viewing window, wherein the viewing is viewing through a microscope.

14. The method of claim 13 , wherein the cell is a muscle cell.

15. The method of claim 13 , wherein moving the first ends of the first and second cell-holding assemblies to the first position results in a stretching force being applied to a region of the cell.

16. The method of claim 13 , wherein the contacting is such that the electrical integrity and electrical functions of the cell, including electrical excitability, is preserved.

17. The method of claim 16 , wherein the cell is a nerve cell.

Assignees

• Univ Rush Medical Center
• Loyola Univ Chicago

Inventors

• Rios Eduardo
• Caron Peter
• Detombe Pieter
• Manno Carlo

Classifications

• C12M29/10

  Perfusion · CPC title

• G02B21/34

  Microscope slides, e.g. mounting specimens on microscope slides · CPC title

• C12M41/36

  of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404) · CPC title

• G02B21/0004

  specially adapted for specific applications · CPC title

• C12M35/02

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title