Method of finding, recording and evaluating object structures

• US 6,529,271 B1
• Filed: 12/20/1999
• Issued: 03/04/2003
• Est. Priority Date: 12/18/1998
• Status: Expired due to Term

• Current Assignee: Leica Microsystems Heidelberg GmbH (Danaher Corporation)
• Original Assignee: Leica Microsystems Heidelberg GmbH (Danaher Corporation)
• Inventors: Engelhardt, Johann
• Primary Examiner(s): EVANS, FANNIE L
• Application Number:
  US09/467,801
• Time in Patent Office: 1,170 Days
• Field of Search: 356/317, 356/318, 356/417, 356/614, 250/458.1, 250/459.1, 250/461.1, 250/461.2
• US Class Current: 356/317
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/008 Details of detection or image processing, including general computer control
• CPC Class Codes: G06T 7/00 Image analysis

Abstract

A method of finding, recording and optionally evaluating object structures, especially on slides, preferably of fluorescent object structures such as gene spots. A microscope with a CCD camera, a scanning microscope or a preferably confocal laser scanning microscope can be used for recording and for the rapid and reliable detection of the object structures, with the image data being recorded using an illumination pattern that is projected into the object plane.

Claims

1. A method of acquiring object structures of an object, comprising:

projecting an illumination pattern onto an object plane where the object is located;

detecting an object structure of the object with a microscope having a light source and a detector;

recording image data corresponding to said object structure with said microscope;

measuring one or more intensity profiles of said object structure during said recording, wherein said intensity profiles are described mathematically by a convolution of said illumination pattern with said object structure; and

cross-correlating said illumination pattern and a shape of said object structure to determine said one or more intensity profiles.

2. A method of acquiring object structures of an object, comprising:

projecting an illumination pattern onto an object plane where the object is located;

detecting an object structure of the object with a microscope having a light source and a detector;

recording image data corresponding to said object structure with said microscope;

measuring one or more intensity profiles of said object structure during said recording, wherein said intensity profiles are described mathematically by a convolution of said illumination pattern with said object structure; and

determining a position of said object structure.

3. A method of acquiring object structures of an object, comprising:

projecting more than one illumination pattern onto an object plane where the object is located; and

detecting an object structure of the object with a microscope having a light source and a detector, wherein at least two illumination sources are used with at least two illumination masks, and wherein an image recording is paralleled using at least two illumination sources.

4. The method according to claim 3, wherein image data is recorded simultaneously with illuminating light of at least two different wavelengths, wherein image data is detected in at least two different detection channels.

5. The method according to claim 4, wherein a mathematical deconvolution on a respective detection channel is carried out with the corresponding illumination mask.

6. The method according to claim 3, further comprising:

implementing a real time data processing with the aid of a dedicated hardware device, said hardware device selected from the group consisting of a digital signal processor (DSP), a field programmable gate array (FPGA), and an application specific integrated circuit (ASIC), such that only extracted object information from the object structures needs to be stored on a data storage unit coupled to said microscope.

7. A method of finding, recording and evaluating fluorescent gene spots on slides, using a microscope with a CCD camera, a scanning microscope or a confocal laser scanning microscope, wherein image data is recorded using an illumination pattern projected into an object plane, and wherein at least two illumination sources are used with identical or different illumination masks.

8. A method of finding, recording and evaluating fluorescent gene spots on slides, using a microscope with a CCD camera, a scanning microscope or a confocal laser scanning microscope, wherein image data is recorded using an illumination pattern projected into an object plane, and wherein an image recording is paralleled using at least two illumination sources.

9. A method of finding, recording and evaluating fluorescent gene spots on slides, using a microscope with a CCD camera, a scanning microscope or a confocal laser scanning microscope, wherein image data is recorded using an illumination pattern projected into an object plane, and wherein intensity profiles measured over the object structures during the recording are described mathematically by convolution of the illumination pattern with the respective object structures.

10. The method as claimed in claim 9, wherein the shape of the intensity profiles is used, by means of suitable computing routines, to detect whether a scan ran centrally or at the edge of the object structures.

11. The method as claimed in claim 9, wherein the intensity profiles are used to determine the actual position of the individual object structures.

12. A method of finding, recording and evaluating fluorescent gene spots on slides, using a microscope with a CCD camera, a scanning microscope or a confocal laser scanning microscope, wherein image data is recorded using an illumination pattern projected into an object plane, wherein information about the object structures recorded is extracted with the aid of mathematical deconvolution of measured image data with the illumination pattern, and wherein image data is recorded simultaneously with illuminating light of at least two different wavelengths, and can be detected in at least two different detection channels.

13. The method as claimed in claim 12, wherein the mathematical deconvolution on the respective detection channel is carried out with the corresponding illumination mask.

14. A method of finding, recording and evaluating fluorescent gene spots on slides, using a microscope with a CCD camera, a scanning microscope or a confocal laser scanning microscope, wherein image data is recorded using an illumination pattern projected into an object plane, wherein illumination and detection are carried out simultaneously via at least two illumination and detection masks, and wherein individual illumination and detection masks have different shapes and sizes.

15. The method as claimed in claim 14, wherein on the basis of a predefined shape of the illumination masks, a mathematical deconvolution is implemented in real time with the aid of a dedicated hardware device, said hardware device selected from the group consisting of a digital signal processor (DSP), a field programmable gate array (FPGA), and an application specific integrated circuit (ASIC), such that only the extracted object information from the object structures is to be stored.