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GB2047884A - Measuring Ion Concentration in Organic Tissue - Google Patents

Measuring Ion Concentration in Organic Tissue Download PDF

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Publication number
GB2047884A
GB2047884A GB8008824A GB8008824A GB2047884A GB 2047884 A GB2047884 A GB 2047884A GB 8008824 A GB8008824 A GB 8008824A GB 8008824 A GB8008824 A GB 8008824A GB 2047884 A GB2047884 A GB 2047884A
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United Kingdom
Prior art keywords
channel
tissue
light
point
solution
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Application number
GB8008824A
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GB2047884B (en
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Science Union et Cie
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Science Union et Cie
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Publication of GB2047884B publication Critical patent/GB2047884B/en
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Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/1455Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using optical sensors, e.g. spectral photometrical oximeters
    • A61B5/1459Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using optical sensors, e.g. spectral photometrical oximeters invasive, e.g. introduced into the body by a catheter
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/1468Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using chemical or electrochemical methods, e.g. by polarographic means
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76Chemiluminescence; Bioluminescence
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/1455Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using optical sensors, e.g. spectral photometrical oximeters
    • A61B5/14551Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using optical sensors, e.g. spectral photometrical oximeters for measuring blood gases
    • A61B5/14556Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using optical sensors, e.g. spectral photometrical oximeters for measuring blood gases by fluorescence
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/40Detecting, measuring or recording for evaluating the nervous system
    • A61B5/4076Diagnosing or monitoring particular conditions of the nervous system
    • A61B5/4094Diagnosing or monitoring seizure diseases, e.g. epilepsy

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Surgery (AREA)
  • Molecular Biology (AREA)
  • Medical Informatics (AREA)
  • Optics & Photonics (AREA)
  • Biophysics (AREA)
  • Biomedical Technology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Plasma & Fusion (AREA)
  • Immunology (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
  • Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)

Abstract

Apparatus for detecting changes in the inorganic ion concentration in organic tissue comprises a tapered hollow support 5 for three channels, channel 1 being adapted to pass light or a light emitting substance into the tissue, channel 2 to pass light from the tissue and channel 3 to contain an electrically conductive saline solution, channel 2 is a bundle of optical fibres leading to photomultiplier 7, and channel 3 contains metal electrode 8 connected to an oscilloscope via an impedance adaptor. The substance carried by channel 1 will generally be one which emits light in the presence of particular cations. <IMAGE>

Description

SPECIFICATION Apparatus for Detecting Changes in inorganic Ion Concentration in Organic Tissue This invention relates to apparatus which may be used to detect continuously, in situ and with a high degree of sensitivity, variations in the concentration of certain ions and molecules in living tissue, and more especially rapid variations in the concentration of intra-organic inorganic cations by means of light emission.
The present invention provides apparatus for use in the detection of changes in the inorganic ion concentration in organic tissue, including first, second and third channels which converge generally towards a point, the first channel being adapted to pass light or a light-emitting substance into the tissue, the second channel being adapted to pass light from the tissue, and the third channel being adapted to contain an electrically-conductive saline solution.
When the first channel is adapted to carry a light-emitting substance, it is suitably provided at its end emote from the point with a device which permits a solution of the light-emitting substance to be injected into the tissue at a predetermined constant pressure so that the solution penetrates the tissue slowly by diffusion. The first channel may be, for example a glass micro-pipette.
The light-emitting substance will generally be a material which emits light in the presence of one or more particular inorganic cations, the quantity of light emitted being dependent upon the concentration of the ion(s) in the tissue.
Alernatively, the first channel is adapted to carry light into the tissue, in which case the channel suitably comprises a quartz optic fiber. In this manner, radiation, for example UV light, may be transmitted to the tissue to excite a particular substance within the tissue to fluoresce, the degree of fluorescence depending upon the concentration of that substance within the tissue.
The second channel which is adapted to carry light away from the tissue suitably comprises an optical fiber which can be connected to a photomultiplier cell and to an appropriate monitoring and/or recording device.
The third channel is adapted to contain an electrically-conductive saline solution, and enables the electrical activity within the tissue under investigation, to be measured. Such activity is suitably recorded by connecting an electrode of an inner material which contacts the saline solution with an oscilloscope via an impedance adaptor.
One form of apparatus according to the present invention is shown schematically by way of example only in Figure 1 of the accompanying drawings.
Referring to the drawing, there are shown three channels 1, 2 and 3 which taper and converge at a point 4. The channels are supported side-byside by a member 5 and may be similarly arranged or arranged in mutual contact at point 4.
The channel 1 through which is injected a solution of a light-emitting substance into the tissue, terminates at its end remote from point 4 in a device 6 which allows the solution to be injected at a fixed constant pressure sufficient to cause the liquid to penetrate by diffusion slowly into the tissue.
The channel 2 is in the form of an optic fiber which conducts out of the tissue the light resulting from the chemical reaction between the light-emitting substance and the ions present at the site of the injection, and is connected by means of a light-conducting bundle of fibers to a photo-multiplier cell 7 bymeans of which it is possible to amplify the intensity of the luminous reaction produced at the site of the injection.
The optic fiber, which may be of quartz, glass or a plastics material, comprises a core which has a high refractive index, and an enveloping layer which is of considerably lower refractive index, so that only the light emitted at the point may be transmitted to the photomultiplier cell. The optic fiber terminates in a frusto-conical point, produced by hot drawing and broken under microscopic control in such a way that the diameter of its point is between 1 and 5 microns.
The channel 3 also has a tapered end and a generally conical body which acts as a reservoir for a saline solution. Into the reservoir there is inserted an inert metal electrode 8 connected to an oscillograph scope via an impedance adaptor.
The channel 3 is most uitably a conventional recording micro-pipette.
Using this apparatus, it is possible simultaneously to monitor and record light emission and changes in potential.
The total length of the apparatus is suitably from 5 to 10 cm, and the diameter at its point is suitably from 2 to 20 /t. Its diameter at the centre will vary depending upon the precise purpose for which it is used.
In a preferred apparatus according to the present invention, the channel 1 has a point diameter of 10 zt and is filled with an aqueous solution of a light-emitting substance and the end of the channel remote from the point 4 has a micrometric plunger or screw or an electro-valve producing a constant injection pressure of the order of 20 cm H20. The optic fiber 2 is connected to a photo-multiplier cell of the cathodic type, by means of an aligning device, optical transmission being facilitated by siliconised lubricant.
The channel 3 forms a reservoir for a 2.5 M solution of sodium chloride or for any other electrically-conductive saline solution. The electrode which is immersed in the reservoir is an electrode or needle of chlorinated silver and is connected to a micro-galvanometer.
The apparatus according to the present invention is especially useful for illustrating changes in the concentration of Ca++ ions at the cellular level, especially in the cortical structure. It has been found that a protein from the luminous Medusa (Aequoria) emits light in the presence of calcium ions and that the quantity of photons produced is a function of the ionic concentration of calcium in a certain biological medium.
Thus, when the channel 1 is filled with an Aequorine solution (*Sigma) and the solution is injected at a constant pressure, a luminous reaction occurs in the tissue which is detected and measured by the photo-multiplier cell connected to the optic fiber 2.
The electrode immersed in the saline solution in the channel 3 enables simultaneous detection of the electrical phenomena accompanying the luminous reaction.
Tests carried out with such apparatus have shown that the detection efficiency for the luminous reaction is approximatley 70%.
The apparatus according to the present invention makes it possible to detect and record very rapid changes in the concentration of Cat+ ions in the cerebral cortex under the influence of a chemical stimulus capable of causing an epileptic fit of the tonic or clonic type, for example penicillin.
During a quite rest period or during deep sleep, the ionic concentration of calcium in the cerebral cortex remains constant. The application of penicillin rapidly induces typical epileptic discharges followed by electrical shocks.
Figure 2 of the accompanying drawings shown graphically results obtained using apparatus according to the invention for determining the changes which occur in the calcium ion concentration in the occipital cortex during an epileptic fit.
Referring first to Figure 2A, the upper plot is of the light signal obtained corresponding to variations in Ca++ activity, expressed in terms of the concentration of calcium ions, against time; the lower plot is the simultaneous electrocorticogram (potential E, against time).
As can be seen from Figure 2A, each isolated epileptic peak is accompanied by significant changes in the concentration of Ca++ which generally follow a sequence of three distinct phases. There is a first phase of reduction in the ionic concentration of calcium before the discharge, then a noticeable increase in Ca++ at the beginning of the peak and a third phase of reduction in the concentration of Ca++ again in the final phase of the peak.
Figures 2B and 2C shown on an enlarged scale details of intercrisis and crisis peaks, respectively.
During the epileptic fit, the concentration of Ca++ undergoes continual changes, and Figure 2C shows the characteristics of these changes and especially a clear increase in calcium in the peak phase.
The apparatus according to the invention therefore offers the advantage of very rapid measurement or detection of changes in ion concentration and a high degree of sensitivity to intracellular ionic currents.
The apparatus according to the invention may also be used for the detection of other neurobiological reactions of the central nervous system by also employing a light-emitting or fluorescent or light-absorbing chemical reaction.
It is therefore possible also to detect with a fluoroescimeter for example, the presence of catecholamine, for example adrenalin or serotonin, or of nicotinamide-reduced adenine diphosphate (NADHz).
It is also possible to detect variations in the concentration of magnesium, using luciferin and luciferase as photo-chemical reagent.
As mentioned above, the first channel may comprise a quartz optic fiber by means of which UV light can be transmitted into the tissue under examination. Under these conditions the light received and transmitted by the second optic fiber corresponds to the fluorescence of the compounds of the organism excited by the ultraviolet light. In this case, the other end of the second optic fiber is connected to a photomultiplier by way of a conventional commercial differential spectro-photometer which makes it possible to analyse the percentage of light emission as a function of the wavelength peculiar to the substance being studied, with reference to a non-specific activity.
The electrical output signals of the photomultiplier and of the electrode in the saline are then passed to a recording and reading amplification system for scientific or industrial application, for example in research into maniacogenic or toxic organic substances in the tissue or the search for indolic substances in tissue sections.
The apparatus according to the invention may also be used in industrial environments. For example, the first conduit may contain a substance which reacts specifically with an ionic component, for example arsenazo, the light absorption of which depends on the local concentration of Ca++. There is then included in the apparatus an additional optic fiber connected to a light source by a suitable monochromator.
This apparatus, connected to a spectrophotometer, makes it possible to measure variations in local concentrations of ions or biological molecules, using a reagent in which the light absorption varies with the concentration of the ion or molecule in question.

Claims (10)

Claims
1. Apparatus for use in the detection of changes in the inorganic ion concentration in organic tissue comprising a tapered hollow support which supports first, second and third channels which converge generally towards a point, the first channel being adapted to pass light or a light-emitting substance into the tissue, the second channel being adapted to pass light from the tissue, and the third channel being adapted to contain an electrically-conductive saline solution.
2. Apparatus according to claim 1, wherein the hollow support is open at its narrower end.
3. Apparatus according to claim 1 or claim 2, wherein the three channels are joined at their ends to open end of the tapered hollow support.
4. Apparatus according to any one of claims 1 to 3, wherein the first channel includes at its end remote from the point, means for injecting liquid into the tissue at a constant fixed pressure.
5. Apparatus according to any one of claims 1 to 4, wherein the first channel contains a solution which reacts with calcium or magnesium ions.
6. Apparatus according to claim 5, wherein the first channel contains a solution of aequorine.
7. Apparatus according to any one of claims 1 to 3, wherein the first channel comprises a quartz fiber.
8. Apparatus according to any one of claims 1 to 7, wherein the second channel is an optic fiber comprising a core having a high refractive index and a sheath of considerably lower refractive index, and having a diameter at its point of from 1 to 25 microns.
9. Apparatus according to any one of claims 1 to 8, wherein the third channel is a micro-pipette.
10. Apparatus according to claim 1 substantially as hereinbefore described with reference to Figure 1 of the accompanying drawings.
GB8008824A 1979-03-16 1980-03-14 Measuring ion concentration in organic tissue Expired GB2047884B (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
FR7906681A FR2451185A1 (en) 1979-03-16 1979-03-16 NEW ION CURRENT DETECTION DEVICE

Publications (2)

Publication Number Publication Date
GB2047884A true GB2047884A (en) 1980-12-03
GB2047884B GB2047884B (en) 1983-04-20

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ID=9223182

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GB8008824A Expired GB2047884B (en) 1979-03-16 1980-03-14 Measuring ion concentration in organic tissue

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BE (1) BE882244A (en)
DE (1) DE3009901A1 (en)
FR (1) FR2451185A1 (en)
GB (1) GB2047884B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19513643A1 (en) * 1995-04-04 1996-10-10 Ludolpf A C Dr Med Testing of biochemical and electrophysiological parameters esp. of human body
GB2381578B (en) * 2001-06-12 2004-04-14 Bosch Gmbh Robert Device and method for testing a material

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3040855A1 (en) * 1980-10-30 1982-06-09 Wolfgang Prof. 7500 Karlsruhe Mehlhardt Examining biological effects on foodstuffs of seeds - by measuring intensity of ultra-weak photon radiation in vitro
DE3038255A1 (en) * 1980-10-10 1982-05-19 Wolfgang Prof. 7500 Karlsruhe Mehlhardt Examining biological effects on foodstuffs of seeds - by measuring intensity of ultra-weak photon radiation in vitro
FR2593916B1 (en) * 1986-01-24 1988-05-13 France Etat Armement SPECTROPHOTOMETER FOR DETERMINATION WITHIN A LIVING ORGANISM
DE3845013B4 (en) * 1987-06-30 2006-09-07 Aaron Lewis Effecting accurate micro-manipulating - via a tube with a small tapered end in proximity to a substrate held in vibration free manner

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1389458A (en) * 1962-10-13 1965-02-19 Eschweiler & Co Apparatus for electric gas analysis
US3811777A (en) * 1973-02-06 1974-05-21 Johnson Res Foundation Medical Time-sharing fluorometer and reflectometer
DE2630606A1 (en) * 1976-07-07 1978-01-12 Alois Hoerl Luminescence radiation analyser for living organisms - uses optical light guide and waveguide on screened measuring head

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19513643A1 (en) * 1995-04-04 1996-10-10 Ludolpf A C Dr Med Testing of biochemical and electrophysiological parameters esp. of human body
GB2381578B (en) * 2001-06-12 2004-04-14 Bosch Gmbh Robert Device and method for testing a material
US7113264B2 (en) 2001-06-12 2006-09-26 Robert Bosch Gmbh Apparatus and method for testing a material

Also Published As

Publication number Publication date
BE882244A (en) 1980-09-15
GB2047884B (en) 1983-04-20
FR2451185A1 (en) 1980-10-10
DE3009901A1 (en) 1980-10-02

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PCNP Patent ceased through non-payment of renewal fee