Method and device for 3 dimensional imaging of suspended micro-objects providing high-resolution microscopy

• US 20060126921A1
• Filed: 10/24/2003
• Published: 06/15/2006
• Est. Priority Date: 10/25/2002
• Status: Active Grant

• Current Assignee: Institut Pasteur, Perkinelmer Cellular Technologies Germany GmbH (Revvity, Inc.)
• Original Assignee: Perkinelmer Cellular Technologies Germany GmbH (Revvity, Inc.)
• Inventors: Schnelle, Thomas, Muller, Torsten, Shorte, Spencer
• Granted Patent: US 7,738,695 B2
• Time in Patent Office: Days
• US Class Current: 382/154
• CPC Class Codes: G01N 2015/144 Imaging characterised by its optical setup
• CPC Class Codes: G01N 2015/1445 Three-dimensional imaging, imaging in different image planes, e.g. under different angles or at different depths, e.g. by a relative motion of sample and detector, for instance by tomography
• CPC Class Codes: G01N 2015/1452 Adjustment of focus; Alignment
• CPC Class Codes: G02B 21/22 Stereoscopic arrangements
• CPC Class Codes: G02B 21/367 providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison

Abstract

A method for high-resolution image recording of at least one object with a microscope, includes the steps of: (a) positioning the object in a receptacle being arranged in the optical axis of the microscope, (b) generating at least two first data sets per object which represent intermediate images of the object with at least two different orientations relative to the optical axis of the microscope, wherein the different orientations of the object are provided by moving the object relative to the receptacle, and (c) evaluating the data sets for obtaining quantitative three dimensional information.

Claims

1. A method for high-resolution image recording of at least one object with a microscope, comprising the steps of:

positioning the at least one object in a receptacle arranged in an optical axis of the microscope,

generating at least two first data sets per object, wherein the at least two first data sets represent intermediate images of the at least one object with at least two different orientations relative to the optical axis of the microscope, and the at least two different orientations of the object are provided by moving the at least one object relative to the receptacle, and

evaluating the data sets for obtaining quantitative three dimensional information.

2. The method according to claim 1, wherein said moving of the at least one object relative to the receptacle comprises a translation and/or rotation of the at least one object by an influence of electric field forces.

3. The method according to claim 2, wherein said translation comprises at least one translation parallel and/or perpendicular relative to the optical axis.

4. The method according to claim 2, wherein said rotation comprises at least one rotation with a rotation axis parallel to the optical axis.

5. The method according to claim 2, wherein said rotation comprises at least one rotation with a rotation axis slanted relative to the optical axis.

6. The method according to claim 5, wherein said rotation axis is slanted within an angle range of up to 90°

.

7. The method according to claim 2, wherein said rotation comprises:

a rotation in a continuous mode or for predetermined time periods and angles, and/or

a rotation with changing rotational axes.

8. The method according to claim 2, wherein said rotation is conducted by holding the at least one object at a fixed position by said electric field forces and by rotating the at least one object by optical forces.

9. The method according to claim 1, further comprising steps of generating further intermediate images of the object, each with another focal plane, respectively, wherein each said focal planes is adjusted by scanning an objective of the microscope parallel to the optical axis.

10. The method according to claim 9, wherein said at least two different orientations of the object and said scanning an objective are conducted in an alternating mode.

11. The method according to claim 1, wherein said positioning comprises suspending said at least one object in a liquid in said receptacle.

12. The method according to claim 1, wherein said evaluating of the resulting data sets comprises a procedure intended to remove out-of-focus light and/or reconstruct a three dimensional map/image of the imaged object.

13. The method according to claim 1, wherein said at least one object comprises at least one eukaryotic cell, at least one prokaryotic cell and/or at least one artificial particle.

14. The method according to claim 1, wherein said microscope is used as a fluorescence microscope, a phase contrast microscope, a differential interference contrast microscope or a confocal microscope.

15. An imaging device for high-resolution image recording of at least one object, comprising:

an microscope imaging system with an optical axis,

a receptacle for accommodating said at least one object, said receptacle being arranged in said optical axis, and

a control circuit being arranged for;

(a) generating at least two first data sets per object, wherein said at least two first data sets represent intermediate images of the at least one object with at least two different orientations relative to the optical axis, and (b) evaluating the data sets for obtaining an object image, and

a driving device adapted to move the at least one object relative to the receptacle.

16. The imaging device according to claim 15, wherein said receptacle comprises a chamber of a fluidic microsystem and said driving device comprises microelectrodes arranged at walls of said chamber and connected with said control circuit.

17. The imaging device according to claim 16, wherein said driving device comprises at least three microelectrodes arranged in one plane in said chamber.

18. The imaging device according to claim 17, wherein said driving device comprises at least six microelectrodes arranged in two planes in said chamber.

19. The imaging device according to claim 15, wherein said control circuit comprises a switching box arranged for switching a rotation axis of the at least one object.

20. A method for high-resolution image recording of at least one object with a measuring device with a predetermined measurement field, comprising the steps of:

positioning the at least one object in a receptacle arranged in the measurement field of the measuring device,

generating at least two first data sets per object, wherein the at least two first data sets represent intermediate data of the at least one object with at least two different orientations relative to the measurement field of the measuring device, and the at least two different orientations of the object are provided by moving the at least one object relative to the receptacle, and

evaluating the data sets for obtaining quantitative three dimensional information.

21. The method according to claim 20, wherein said measuring device comprises a microscope and said measurement field is an optical axis of the microscope.

22. The method according to claim 20, wherein said measuring device comprises an impedance measurement device and said measurement field is the receptacle.