CA2387492A1 - Apparatus for measuring an oxygen concentration gradient and method of use thereof - Google Patents
Apparatus for measuring an oxygen concentration gradient and method of use thereof Download PDFInfo
- Publication number
- CA2387492A1 CA2387492A1 CA002387492A CA2387492A CA2387492A1 CA 2387492 A1 CA2387492 A1 CA 2387492A1 CA 002387492 A CA002387492 A CA 002387492A CA 2387492 A CA2387492 A CA 2387492A CA 2387492 A1 CA2387492 A1 CA 2387492A1
- Authority
- CA
- Canada
- Prior art keywords
- sample
- phosphorescence
- signal
- concentration gradient
- excitation light
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
- A61B5/1455—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using optical sensors, e.g. spectral photometrical oximeters
- A61B5/14551—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using optical sensors, e.g. spectral photometrical oximeters for measuring blood gases
- A61B5/14556—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using optical sensors, e.g. spectral photometrical oximeters for measuring blood gases by fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Abstract
The invention provides an improved, reliable and efficient way of measuring the oxygen concentration gradient in a sample by calculating linear oxygen concentrations within the sample, permitting diagnostic testing, for example, of the effects of a developmental or metabolic change in a cell or tissue, in vitro orin vivo, in response to disease, injury, radiation, or mechanical or chemical intervention, or simply to changed circumstances, or to measure the oxygen permeability of a membrane or plastic. The apparatus in a preferred embodiment comprises a core digital signal processor (45) having sufficient memory to perform the necessary calculations, to control output of excitation light from a light source, and to collect phosphorescent lifetime data (41) and signal processors (A/D and D/A) (44, 46).
Claims (17)
1. An apparatus for measuring the oxygen concentration gradient within a phosphor-containing sample in terms of a measured phosphorescence lifetime, wherein the apparatus comprises:
a core digital signal processor (DSP), having sufficient memory (RAM and ROM) to perform the necessary calculations, to control output of excitation light from a light source, and to collect phosphorescent lifetime data;
a first Delta Sigma signal processor (D/A, digital to analog);
an avalanche photodiode or photomultiplier;
an amplifier; and a second Delta-Sigma signal processor (A/D analog to digital) responsive to the amplified output from the photodiode or photomultiplier.
a core digital signal processor (DSP), having sufficient memory (RAM and ROM) to perform the necessary calculations, to control output of excitation light from a light source, and to collect phosphorescent lifetime data;
a first Delta Sigma signal processor (D/A, digital to analog);
an avalanche photodiode or photomultiplier;
an amplifier; and a second Delta-Sigma signal processor (A/D analog to digital) responsive to the amplified output from the photodiode or photomultiplier.
2. The apparatus according to claim 1, further comprising:
a core digital signal processor (DSP), having sufficient memory (RAM and ROM) to perform the necessary calculations, to control output of the excitation source, and to collect phosphorescent lifetime data;
a first Delta Sigma signal processor (D/A, digital to analog) for converting tabulated calculated data to current to control an excitation light signal from the selected light source;
an avalanche photodiode or photomultiplier for filtering and detecting emitted phosphorescent light from the sample following exposure to the excitation light signal;
an amplifier for amplifying the output of the photodiode or photomultiplier; a second Delta-Sigma signal processor (A/D analog to digital) responsive to the amplified output from the photodiode or photomultiplier, for digitizing the amplified photodetector output (the emitted phosphorescence), and for compiling collected data into a separate memory set, m (the tabulated calculated data), in the DSP, wherein data is summed to recover distribution of the phosphorescent lifetimes, from which oxygen concentration gradient is calculated from at least one equation.
a core digital signal processor (DSP), having sufficient memory (RAM and ROM) to perform the necessary calculations, to control output of the excitation source, and to collect phosphorescent lifetime data;
a first Delta Sigma signal processor (D/A, digital to analog) for converting tabulated calculated data to current to control an excitation light signal from the selected light source;
an avalanche photodiode or photomultiplier for filtering and detecting emitted phosphorescent light from the sample following exposure to the excitation light signal;
an amplifier for amplifying the output of the photodiode or photomultiplier; a second Delta-Sigma signal processor (A/D analog to digital) responsive to the amplified output from the photodiode or photomultiplier, for digitizing the amplified photodetector output (the emitted phosphorescence), and for compiling collected data into a separate memory set, m (the tabulated calculated data), in the DSP, wherein data is summed to recover distribution of the phosphorescent lifetimes, from which oxygen concentration gradient is calculated from at least one equation.
3. The apparatus according to claim 2, wherein the data collected by the second signal processor (the digitizer) is synchronized with the first signal processor (the D/A unit) to control the driving current controlling the selected light source.
4. The apparatus according to claim 2, wherein the emitted phosphorescence is functionally related to oxygen quenching when the sample is exposed to the excitation light.
5. An apparatus according to claim 4, wherein the light source introduces a plurality of signals into the sample, such that a set of signals is established in the sample, from which set a waveform is derived, where within said waveform, all component waveforms pass through zero.
6. An apparatus according to claim 1, wherein the photodetector or photomultiplier detects a plurality of emitted signals corresponding to a plurality of excitation signals introduced into the sample as the excitation light, and wherein the detection means determines a solution of at least one equation based upon variations in the respective values of the signal parameters of the plurality of detected emission signals.
7. An apparatus according to claim 4, wherein the predetermined signal parameter is excitation frequency, and wherein said signal source introduces a plurality of signals having different respective excitation frequencies into the sample.
8. An apparatus according to claim 6, wherein all modulation frequencies are mixed within the excitation light.
9. An apparatus according to claim 4, wherein the measured signal parameter is emitted phosphorescence from the phosphor-containing sample exposed to excitation light, wherein the phosphorescence is inversely related to oxygen quenching in the sample.
10. An apparatus according to claim 6, wherein the oxygen concentration gradient is extracted from a dependence of phosphorescence amplitude and phase angle on the modulation frequency in the plurality of detected signals.
11. An apparatus according to claim 1 wherein the second signal processor further comprises:
a means for regularizing the detected phosphorescence signals; and a means, responsive to said regularizing means, for representing the regularized signals by a solution using fast, non-iterative quadratic programming algorithm at each maximizing step to interpolate a histogram representing the best underlying distribution of the phosphorescence lifetimes.
a means for regularizing the detected phosphorescence signals; and a means, responsive to said regularizing means, for representing the regularized signals by a solution using fast, non-iterative quadratic programming algorithm at each maximizing step to interpolate a histogram representing the best underlying distribution of the phosphorescence lifetimes.
12. The apparatus according to claim 11, wherein the histogram representing the best underlying distribution of the phosphorescence lifetimes is converted into a distribution of oxygen concentrations by the Stern-Volmer relationship.
13. The apparatus according to claim 1, wherein one or more elements of the apparatus operate automatically and interconnectively.
14. A method for determining an oxygen concentration gradient in a sample comprising:
dissolving or introducing a hydrophilic phosphorescent compound in the sample, wherein quenching constant and lifetime at zero oxygen are known or previously determined for the phosphorescent compound;
illuminating the sample with a pulsed or modulated excitation light at an intensity and frequency sufficient to cause the phosphorescent compound to emit a measurable phosphorescence;
measuring the emitted phosphorescence; and calculating the phosphorescence lifetime and oxygen concentration gradient in the sample.
dissolving or introducing a hydrophilic phosphorescent compound in the sample, wherein quenching constant and lifetime at zero oxygen are known or previously determined for the phosphorescent compound;
illuminating the sample with a pulsed or modulated excitation light at an intensity and frequency sufficient to cause the phosphorescent compound to emit a measurable phosphorescence;
measuring the emitted phosphorescence; and calculating the phosphorescence lifetime and oxygen concentration gradient in the sample.
15. An article for determining oxygen concentration gradient from detected phosphorescence lifetimes in a phosphor-containing sample based upon a signal that has propagated through at least a portion of the sample, wherein the signal varies with respect to excitation frequencies from an excitation light source and emitted phosphorescence, and wherein the emitted phosphorescence varies in an inverse direct relationship to oxygen quenching in the sample, wherein the article comprises:
a computer-readable storage medium;
means in the medium for analyzing the emitted phosphorescence signal detected from the sample to determine variations in the signal with respect to a predetermined quenching constant and maximal lifetime at zero oxygen for the phosphor, means in the medium for constructing one or more equations at least partially based upon the signal, wherein an equation extracts the dependence of phosphorescence amplitude and phase angle with the summation of modulation frequencies in the excitation light;
means in the medium for determining a solution of the one or more equations which have been constructed to resolve the variations in phosphorescence amplitude and phase angle with respect to modulation frequencies and the quenching constant and maximal lifetime at zero oxygen for the selected phosphor;
means in the medium recovering an algorithmically-determined histogram which maximally resembles the phosphorescence lifetime distribution of the selected phosphor in the sample; and means in the medium for algorithmically-converting the phosphorescence lifetime distribution into a corresponding oxygen concentration gradient based upon the Sterne-Volmer relationship.
a computer-readable storage medium;
means in the medium for analyzing the emitted phosphorescence signal detected from the sample to determine variations in the signal with respect to a predetermined quenching constant and maximal lifetime at zero oxygen for the phosphor, means in the medium for constructing one or more equations at least partially based upon the signal, wherein an equation extracts the dependence of phosphorescence amplitude and phase angle with the summation of modulation frequencies in the excitation light;
means in the medium for determining a solution of the one or more equations which have been constructed to resolve the variations in phosphorescence amplitude and phase angle with respect to modulation frequencies and the quenching constant and maximal lifetime at zero oxygen for the selected phosphor;
means in the medium recovering an algorithmically-determined histogram which maximally resembles the phosphorescence lifetime distribution of the selected phosphor in the sample; and means in the medium for algorithmically-converting the phosphorescence lifetime distribution into a corresponding oxygen concentration gradient based upon the Sterne-Volmer relationship.
16. A method of using the apparatus according to claim 1 to detect phosphorescence lifetimes in the phosphor-containing sample.
17. A method of using the apparatus according to claim 1 to determine the oxygen concentration gradient in the phosphor-containing sample.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US15961699P | 1999-10-14 | 1999-10-14 | |
| US60/159,616 | 1999-10-14 | ||
| PCT/US2000/041171 WO2001027585A2 (en) | 1999-10-14 | 2000-10-13 | Oxygen concentration gradient measurement apparatus and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2387492A1 true CA2387492A1 (en) | 2001-04-19 |
| CA2387492C CA2387492C (en) | 2012-07-10 |
Family
ID=22573286
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2387492A Expired - Fee Related CA2387492C (en) | 1999-10-14 | 2000-10-13 | Apparatus for measuring an oxygen concentration gradient and method of use thereof |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP1220639A4 (en) |
| AU (1) | AU766128B2 (en) |
| CA (1) | CA2387492C (en) |
| WO (1) | WO2001027585A2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1742038A1 (en) | 2005-07-06 | 2007-01-10 | Academisch Medisch Centrum bij de Universiteit van Amsterdam | Device and method for determining the concentration of a substance |
| EP2318823B1 (en) | 2008-08-15 | 2015-12-02 | Erasmus University Medical Center Rotterdam | Methods and devices for assessment of mitochondrial function |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5186173A (en) * | 1990-08-14 | 1993-02-16 | Drexel University | Method for in vivo measurement of oxygen concentration levels |
| US5281825A (en) * | 1991-09-05 | 1994-01-25 | The University Of Maryland School Of Medicine | Phase fluorometry using a modulated electroluminescent lamp as a light source |
| US5462879A (en) * | 1993-10-14 | 1995-10-31 | Minnesota Mining And Manufacturing Company | Method of sensing with emission quenching sensors |
| US5515864A (en) * | 1994-04-21 | 1996-05-14 | Zuckerman; Ralph | Method and apparatus for the in vivo measurement of oxygen concentration levels by the indirect determination of fluoescence lifetime |
| US5515694A (en) * | 1995-01-30 | 1996-05-14 | Carrier Corporation | Subcooler level control for a turbine expansion refrigeration cycle |
| US5863460A (en) * | 1996-04-01 | 1999-01-26 | Chiron Diagnostics Corporation | Oxygen sensing membranes and methods of making same |
| US5830138A (en) * | 1996-12-16 | 1998-11-03 | Trustees Of The University Of Pennsylvania | Intravascular catheter probe for clinical oxygen, pH and CO2 measurement |
| US6165741A (en) * | 1997-05-30 | 2000-12-26 | The Trustees Of The University Of Pennsylvania | Method for rapid detection of bacterial growth in cultures |
-
2000
- 2000-10-13 AU AU19649/01A patent/AU766128B2/en not_active Ceased
- 2000-10-13 EP EP00982645A patent/EP1220639A4/en not_active Withdrawn
- 2000-10-13 CA CA2387492A patent/CA2387492C/en not_active Expired - Fee Related
- 2000-10-13 WO PCT/US2000/041171 patent/WO2001027585A2/en active IP Right Grant
Also Published As
| Publication number | Publication date |
|---|---|
| EP1220639A4 (en) | 2006-06-07 |
| CA2387492C (en) | 2012-07-10 |
| AU1964901A (en) | 2001-04-23 |
| WO2001027585A2 (en) | 2001-04-19 |
| EP1220639A2 (en) | 2002-07-10 |
| AU766128B2 (en) | 2003-10-09 |
| WO2001027585A3 (en) | 2002-01-03 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20141014 |