Method and a system for determination of particles in a liquid sample

• US 8,432,550 B2
• Filed: 03/28/2012
• Issued: 04/30/2013
• Est. Priority Date: 05/05/1997
• Status: Expired due to Fees

• Current Assignee: ChemoMetec A/S
• Original Assignee: ChemoMetec A/S
• Inventors: Hansen, Frans Ejner Ravn, Jeppesen, Jesper Myron, Glensbjerg, Martin, Arnvidarson, Brkur
• Primary Examiner(s): Toatley, Gregory J
• Assistant Examiner(s): MERLINO, AMANDA H
• Application Number:
  US13/433,010
• Publication Number:
  US 20120223260A1
• Time in Patent Office: 398 Days
• Field of Search: None
• US Class Current: 356/436
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/025 Methods for single or grouped particles
• CPC Class Codes: G01N 2015/1452 Adjustment of focus; Alignment
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2021/5957 using an image detector type detector, e.g. CCD
• CPC Class Codes: G01N 2021/6491 Measuring fluorescence and transmission; Correcting inner filter effect
• CPC Class Codes: G01N 21/31 Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry G01N21/72 takes precedence
• CPC Class Codes: G01N 21/59 Transmissivity G01N21/25 takes precedence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/04 Dairy products

Abstract

A method for the assessment of quantity and quality parameters of biological particles in a liquid analyte material. The method comprises applying a volume of a liquid sample to an exposing domain from which exposing domain electromagnetic signals from the sample in the domain can pass to the exterior, and exposing, onto an array of active detection elements such as CCD-elements, a spatial representation of electromagnetic signals having passed from the domain, the representation being detectable as an intensity by individual active detection elements, under conditions permitting processing of the intensities detected by the array of detection elements during the exposure in such a manner that representations of electromagnetic signals from the biological particles are identified as distinct from representations of electromagnetic signals from background signals.

Claims

1. A system for the assessment of at least one quantity parameter and/or at least one quality parameter of biological particles in a liquid analyte material, comprising

a sample compartment having a wall part defining an exposing area, the wall part allowing electromagnetic radiation from a liquid sample in the sample compartment to pass through the wall and to be exposed to the exterior, the volume of the liquid sample from which electromagnetic radiation is exposed onto an array of active detection elements being in the range of 0.01 μ

l to 20 μ

l,

one or more LEDs arranged to transmit light onto the sample,

an array of active detection elements, onto which an at least one-dimensional spatial image of said volume can be exposed, the image being one which is detectable as an intensity by individual active detection elements, under conditions which will permit processing of the intensities detected by the array of detection elements during the exposure in such a manner that images of electromagnetic signals from the biological particles are identified as distinct from images of electromagnetic signals from background signals,

a lens consisting of one or several elements for focusing an image of electromagnetic signals from at least part of the exposing domain onto the array of detection elements, the spatial representation exposed onto the array of detection elements being subject to such a linear enlargement that the ratio of a linear dimension of the image on the array of detection elements to the original linear dimension in the exposing domain is smaller than 10;

1,

at least one monochromatic device for separating electronic radiation into substantially one or more wavelength components or wavebands allowing the system to receive fluorescence, and

a computer unit for processing the intensities detected by the detection elements in such a manner that signals from the biological particles are identified as distinct from background signals, and for correlating the results of the processing to the at least one quantity parameter and/or the at least one quality parameter of the liquid analyte material.

2. The system according to claim 1, wherein at least a major part of the electromagnetic radiation from the light source, has a direction which is transverse to the wall of the sample compartment or a plane defined by the domain.

3. The system according to claim 1, comprising a plurality of light emitting diodes.

4. The system according to claim 3, wherein the light emitting diodes emit electromagnetic radiation of substantially the same wavelength.

5. The system to claim 3, wherein at least two light emitting diodes have substantially different radiation properties in at least one waveband, the light emitting diodes being operated in such a way that all transmit substantially simultaneously or at least one source transmits while at least one diode is not transmitting, or at least one diode is transmitting at a time.

6. The system of claim 1, comprising one or more valves which can control the flow of sample or any other component.

7. The system according to claim 1, wherein the array of detection elements is arranged in two directions in such a way that the detection elements form a series of substantially parallel straight lines, the series forming a rectangle.

8. The system according to claim 1, wherein the ratio is at the most 6:

1.

9. The system according to claim 1, wherein the ratio is smaller than 4.1.

10. The system according to claim 1, wherein the ratio is in the range between 10:

1 and 1;

10.

11. The system according to claim 1, wherein the ratio is in the range between 1:

1 and 1;

100.

12. The system according to claim 1, wherein the interior of the sample compartment has an average thickness of 20 μ

m to 200 μ

m.

13. The system according to claim 1, wherein the sample compartment has dimensions, in a direction substantially parallel to the array of detection elements, in the range between 1 mm by 1 mm and 10 mm by 10 mm.

14. The system according to claim 1, wherein the volume of the liquid sample from which electromagnetic radiation is exposed onto the array is in the range of 0.01 μ

l to 1 μ

l.

15. The system according to claim 1, wherein the volume of the liquid sample from which electromagnetic radiation is exposed onto the array is in the range of 0.04 μ

l to 4 μ

l.

16. The system according to claim 1, comprising a pump, said pump being situated either upstream to the sample compartment or downstream to the sample compartment.

17. The system according to claim 16, wherein the pump is one or several of the following:

peristaltic pump piston pump, membrane pump, centrifugal pump, and hypodermic syringe.

18. The system according to claim 1, wherein the computer unit is capable of assessing the number of the biological particles per volume of the liquid analyte material.

19. The system according to claim 1, wherein the computer unit is capable of assessing the size and/or shape of the biological particles in the liquid analyte material.

20. The system according to claim 1, wherein the computer unit is capable of assessing is the presence or non-presence of a particular type of particles in the liquid analyte material.

21. The system according to claim 1, wherein the duration of the exposure is in the range from 100 milliseconds to 5 seconds.

22. The system according to claim 1, wherein the exposure is performed as a single exposure.

23. The system according to claim 1, where said array of detection elements is one or several of the following types:

full frame CCD, frame transfer CCD, interline transfer CCD, line scan CCD, and CMOS image sensor.

24. The system according to claim 1, where the lens has the effect of substantially increasing the intensity of said electromagnetic radiation in or at said sample.

25. The system according to claim 1, further comprising a container for one or more chemical components, the container being connected to a flow system where the sample flows, at least a portion of the sample flowing through the chemical container and thus allowing the mixing of the chemical components with the sample.

26. The system according to claim 1, wherein at least one monochromatic device is selected from interference filters, colored filters, an optical grating, a prism, and optionally active crystals.

27. The system according to claim 1, wherein the sample compartment is a removable sample compartment, which can be readily removed from the system.

28. The system according to claim 1, wherein the sample compartment comprises one or more compartments, where chemical or physical components can be stored such that the chemical or physical components can be added to any sample present in the sample compartment one at a time or more than one at a time.

29. The system according to claim 1, comprising at least 2 of the following:

interference filters, colored filters, an optical grating, a prism, and optically active crystals.

30. The system according to claim 1, being adapted to keep the sample in the sample compartment at stand still during the exposure.