ANALYSIS METHOD FOR CHEMICAL AND/OR BIOLOGICAL SAMPLES

• US 20090290780A1
• Filed: 07/17/2007
• Published: 11/26/2009
• Est. Priority Date: 07/17/2006
• Status: Abandoned Application

• Current Assignee: Perkinelmer Cellular Technologies Germany GmbH (Revvity, Inc.)
• Original Assignee: Perkinelmer Cellular Technologies Germany GmbH (Revvity, Inc.)
• Inventors: Kottig, Karsten
• Application Number:
  US12/309,445
• Publication Number:
  US 20090290780A1
• Time in Patent Office: Days
• US Class Current: 382/133
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16

Abstract

An analysis method for chemical and/or biological samples, particularly chemical and/or biological samples comprising cells, includes the following steps:

• taking a sample image (46), said sample image (46) comprising a plurality of pixels,
• generating analysis data per pixel,
• determining pixels of interest for the analysis, and
• evaluating the generated analysis data per pixel of interest, preferably by a fluctuation analysis procedure,
  and is characterized in that said analysis data are generated during said taking of the sample image and comprise pixel information resolved into time series, said pixel information being used for evaluation preferably on the basis of a fluctuation analysis procedure.

Claims

1. An analysis method for chemical and/or biological samples, particularly chemical and/or biological samples comprising cells, said method including the following steps:

taking a sample image, said sample image comprising a plurality of pixels,

generating analysis data per pixel,

determining pixels of interest for the analysis, and

evaluating the generated analysis data per pixel of interest, preferably by fluctuation analysis procedures,

whereby said analysis data are generated during said taking of the sample image and comprise pixel information resolved into time series, said pixel information being used for evaluation preferably on the basis of a fluctuation analysis procedure and whereby the analysis data of a plurality of pixels of interest are combined into group analysis data to be evaluated together.

2. The analysis method according to claim 1, wherein, in a sample comprising at least one, preferably a multitude of cells, pixel types are determined in dependence on subcellular structures.

3. The analysis method according to claim 2, wherein the analysis data of pixel types are combined into group analysis data to be evaluated together.

4. The analysis method according to claim 1, wherein the pixels of interest are determined with the aid of a threshold analysis.

5. The analysis method according to claim 1, wherein the fluctuation analysis procedure comprises fluorescence correlation spectroscopy, one- or multi-dimensional fluorescence intensity distribution analysis, fluorescence intensity multiple distribution analysis, fluorescence intensity lifetime distribution analysis, one- or multi-dimensional photon counting histogram analysis, photon arrival-time interval distribution analysis, and/or burst-integrated fluorescence lifetime analysis.

6. The analysis method according to claim 1, wherein the analysis data of a plurality of pixels of interest are combined into group analysis data by adding the time series acquired for multiple pixels.

7. The analysis method according to claim 1, wherein the analysis data of a plurality of pixels of interest are combined into group analysis data by:

for each pixel, combining the time series into a histogram per pixel, and

combining said histograms into a group histogram.

8. The analysis method according to claim 1, wherein the analysis data of a plurality of pixels of interest are combined into group analysis data by:

for each pixel, combining the time series into an autocorrelation function per pixel, and

evaluating said autocorrelation functions together, preferably by combined fitting of said autocorrelation functions.

9. The analysis method according to claim 8, wherein the pixel traces have been generated beforehand by mask generation, particularly with the aid of cell recognition routines.

10. The analysis method according to claim 1, wherein said time series is resolved into individual time segments and wherein, in said time segments, preferably the arrival of photons is observed.

11. The analysis method according to claim 1, wherein the analysis data of a pixel comprise individual data which are taken per time segment and particularly are stored per time segment.

12. The analysis method according to claim 11, wherein the numbers of arrived photons per time segment are detected as said individual data.

13. The analysis method according to claim 11, wherein the temporal intervals between arrived photons are detected as said individual data, the detection being preferably performed within time segments.

14. The analysis method according to claim 11, wherein said time segments have a length of 100 ns to 10 ms.

15. The analysis method according to claim 1, wherein the time for taking a sample image is 0.1 to 100 s.

16. The analysis method according to claim 1, wherein the determining of the pixels of interest is performed temporally after the generating of the analysis data.