NZ730251B2 - Device for real-time in vivo molecular analysis - Google Patents
Device for real-time in vivo molecular analysisInfo
- Publication number
- NZ730251B2 NZ730251B2 NZ730251A NZ73025115A NZ730251B2 NZ 730251 B2 NZ730251 B2 NZ 730251B2 NZ 730251 A NZ730251 A NZ 730251A NZ 73025115 A NZ73025115 A NZ 73025115A NZ 730251 B2 NZ730251 B2 NZ 730251B2
- Authority
- NZ
- New Zealand
- Prior art keywords
- laser
- capillary
- mass spectrometer
- transfer tube
- analysis
- Prior art date
Links
- 238000007479 molecular analysis Methods 0.000 title claims abstract description 8
- 238000001727 in vivo Methods 0.000 title description 21
- 238000004458 analytical method Methods 0.000 claims abstract description 49
- 238000012546 transfer Methods 0.000 claims abstract description 49
- 239000000523 sample Substances 0.000 claims abstract description 36
- 239000000835 fiber Substances 0.000 claims abstract description 25
- 239000012620 biological material Substances 0.000 claims abstract description 24
- 239000002245 particle Substances 0.000 claims abstract description 24
- 230000003287 optical effect Effects 0.000 claims abstract description 5
- 238000002679 ablation Methods 0.000 claims description 15
- 239000002904 solvent Substances 0.000 claims description 13
- 238000002560 therapeutic procedure Methods 0.000 claims description 10
- 150000001793 charged compounds Chemical class 0.000 claims description 8
- 238000005286 illumination Methods 0.000 claims description 7
- 229910052751 metal Inorganic materials 0.000 claims description 7
- 239000002184 metal Substances 0.000 claims description 7
- 239000000463 material Substances 0.000 abstract description 10
- 238000000034 method Methods 0.000 description 38
- 150000002500 ions Chemical class 0.000 description 29
- 239000012071 phase Substances 0.000 description 11
- 210000000056 organ Anatomy 0.000 description 10
- 230000008901 benefit Effects 0.000 description 8
- 238000004949 mass spectrometry Methods 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 7
- 238000000688 desorption electrospray ionisation Methods 0.000 description 7
- 230000001965 increasing effect Effects 0.000 description 7
- 239000007789 gas Substances 0.000 description 6
- 210000004185 liver Anatomy 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 238000002663 nebulization Methods 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 238000005040 ion trap Methods 0.000 description 4
- 238000000608 laser ablation Methods 0.000 description 4
- 238000000091 laser ablation electrospray ionisation Methods 0.000 description 4
- 238000002647 laser therapy Methods 0.000 description 4
- 230000007170 pathology Effects 0.000 description 4
- 239000004809 Teflon Substances 0.000 description 3
- 229920006362 Teflon® Polymers 0.000 description 3
- 238000001574 biopsy Methods 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000001698 laser desorption ionisation Methods 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000010223 real-time analysis Methods 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 229910003327 LiNbO3 Inorganic materials 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000003795 desorption Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000005684 electric field Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 150000004665 fatty acids Chemical class 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 229910000040 hydrogen fluoride Inorganic materials 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- XUKUURHRXDUEBC-KAYWLYCHSA-N Atorvastatin Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-KAYWLYCHSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 208000001654 Drug Resistant Epilepsy Diseases 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 1
- 229910052689 Holmium Inorganic materials 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 238000001069 Raman spectroscopy Methods 0.000 description 1
- 229910052769 Ytterbium Inorganic materials 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 229910052797 bismuth Inorganic materials 0.000 description 1
- JCXGWMGPZLAOME-UHFFFAOYSA-N bismuth atom Chemical compound [Bi] JCXGWMGPZLAOME-UHFFFAOYSA-N 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000002828 effect on organs or tissue Effects 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000007792 gaseous phase Substances 0.000 description 1
- 230000007274 generation of a signal involved in cell-cell signaling Effects 0.000 description 1
- KJZYNXUDTRRSPN-UHFFFAOYSA-N holmium atom Chemical compound [Ho] KJZYNXUDTRRSPN-UHFFFAOYSA-N 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 238000010249 in-situ analysis Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000009616 inductively coupled plasma Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000002595 magnetic resonance imaging Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 239000004810 polytetrafluoroethylene Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001004 secondary ion mass spectrometry Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- NAWDYIZEMPQZHO-UHFFFAOYSA-N ytterbium Chemical compound [Yb] NAWDYIZEMPQZHO-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/02—Instruments for taking cell samples or for biopsy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
-
- 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
- A61B5/14503—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 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
-
- 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
- A61B5/14546—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 for measuring analytes not otherwise provided for, e.g. ions, cytochromes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N2001/045—Laser ablation; Microwave vaporisation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/566—Immunoassay; Biospecific binding assay; Materials therefor using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/02—Details
- H01J49/04—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components
- H01J49/0459—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components for solid samples
- H01J49/0463—Desorption by laser or particle beam, followed by ionisation as a separate step
Abstract
The invention relates to a biological material molecular analysis device characterized in that it includes: - a laser (34) optionally using an optical parametric oscillator (OPO), configured to emit a wavelength between 2.5 ym and 12 put, said configured laser (34) being intended to ablate said biological material by ejecting charged and/or uncharged particles; - a mass spectrometer (31); and - a probe (S, 10) comprising at least one first analysis fiber (A, 14), connected to the laser (34), and a transfer tube (T, 21), connected to the mass spectrometer (31). ogical material by ejecting charged and/or uncharged particles; - a mass spectrometer (31); and - a probe (S, 10) comprising at least one first analysis fiber (A, 14), connected to the laser (34), and a transfer tube (T, 21), connected to the mass spectrometer (31).
Description
DEVICE FOR REAL TIME IN VIVO MOLECULAR ANALYSIS FIELD The present invention relates to a device for in vivo lar is in real time.
The field of the invention is that of the analysis of cells of a living organism.
BACKGROUND Early diagnosis of pathologies is a crucial step for surgeons and clinicians. The diagnostics should lead to making a clear decision as rapidly as possible on the physiopathological condition of the patient (human or animal). This step should be carried out within a short lapse of time and this with a minimum of damages for the patient and without imposing on him/her additional complications.
For 25 years, several non-invasive diagnostic tools have been developed, notably magnetic resonance imaging, the scanner, Positron Emission aphy (PET) or sinography. These techniques are effective for observing, localizing and determining the size of abnormal regions from a tissue point of view like cancer regions. Certain of these techniques may even give more specific information like the production of new blood vessels inside these regions (neoangiogenesis) or cell catabolism.
However, none of these techniques has the capacity of giving information on the molecular contents of the relevant region. This information is notably absent for posing a diagnostic or even a se of the pathology. Such a widely used strategy within the hospital is to proceed with excision of tissues y) in the abnormal region and then carrying out ex-vivo is on the tissue with different techniques and notably histological techniques (e.g. anatomopathological ation) for seeking morphological, cell, tissue or molecular teristics. Within the scope of cancers, such ce gives the possibility of confirming the presence of malign tumors and to obtain their histological classification (type, . Other more targeted techniques may be applied for obtaining a diagnostic such as immunohistochemistry (IHC) or PCR techniques in order to search for specific markers of the pathology or particular mutations.
Although this gy is widely used, it may prove to be long, g during this period the patient in the operating room while waiting for his/her diagnostic. Therefore there is a real interest for the development of techniques ng the collection of in-vivo molecular information. The sought device should give the possibility of obtaining this invivo information but also in real time during the time of presence in the ing room.
From among the techniques which may give the possibility of obtaining in-vivo molecular information, there is a need for oscopic techniques. Raman, IR or 18244134_1 (GHMatters) P105417.NZ fluorescence oscopies are techniques g these criteria. However, these techniques have certain drawbacks which are either the requirement of using a tracer for viewing the region of interest, or of collecting complex profiles (i.e. each le has in the analyzed region gives a complex spectrum and the spectrum of the analyzed cell region is a superposition of the whole of the spectra of the molecules making up the region) not always giving the possibility of observing molecular variations between a normal and pathological area and requiring the use of extremely complex statistical processing ions.
On the other hand, r oscopic method, mass spectrometry, may meet this need for a rapid diagnostic in-vivo in real time. Mass spectrometry is a technique based on the measurement of the molecular weight of the s. Conventionally, the mass ement is carried out according to the diagram, generation of ions in a gas phase from the sample (in-vitro) by the source of producing ions of the instrument, separation of the formed ions according to the ratio m/z in the analyzer portion and then detection of the ion current. The ed samples may be solid, liquid or gaseous.
However, the source of ions used will be adapted to the condition in which is found the sample. From complex mixtures, mass spectrometry provides the advantage of allowing observation of a signal for each species since the latter are separated according to m/z except if the compounds have the same raw formula or if the performances of the ment are insufficient. Historically, mass spectrometry leads to the arrival of different technologies for sources of ion production and analyzers, the sources and analyzers being able to be combined with each other in different ways allowing the creation of instruments with characteristics defined in terms of compounds which may be analyzed, of sample condition, of instrumental performances. More recently, mass ometry techniques have evolved so as to give the possibility of passing from the analysis of invitro extracts to the analysis of organisms or organism portions ex-vivo. The development of these ques has contributed to the arrival of a new field of research entitled Imaging by Mass Spectrometry. Presently, the sources for producing ions the most currently used since compatible in this field are the s of so called Secondary Ion Mass Spectrometry (SIMS), Laser Desorption Ionization (LDI) sources, the Matrix Assisted Laser Desorption/Ionization (MALDI) sources, the Desorption Electrospray Ionization sources (DESI) and the Laser Ablation-Inductively Coupled Plasma (LA-ICP).
These technologies as such allow analysis of sms or organism portions o for which they give the possibility of molecular characterization but they cannot be used in- vivo on living organisms. 18244134_1 (GHMatters) P105417.NZ Indeed, in the case of DESI "for Desorption Electrospray Ionization", a jet of charged droplets of solvent produced by an electro-nebulization process is directed onto the surface of the sample. The droplets will rebound at the surface of the sample with a process for capturing surface molecules during the interaction phase of the droplets with the surface. The molecules are sucked up by a capillary intended for entering the mass spectrometer. The DESI source has proven its ties for multiple biological samples like tissues or organs. This is illustrated in the article of Calligaris D et al., 2013, J Mass Spectrometry, 48 (II), 1178-87 wherein DESI is combined with conventional in-vivo imaging. An instrumental modification was carried out for attempting the use of DESI in vivo: Chen CH et al, 2013, Anal. Chem 85(24), 11843-50. In this case, a pressurized solvent jet is directed onto the tissue. The jet is positioned inside a transfer tube giving the possibility of ensuring the transport of the charged molecules generated by the jet towards the mass spectrometer. In spite of its use in-vivo, this ment requires a contact with the area to be analyzed. The analysis of the surface in a uous way may induce contamination effect for characterization of biological tissues.
In order to circumvent this problem, a solution is laser ablation as a desorption method: Nemes P, Vertes A, 2007 Anal. Chem., 79(21), 8098-106 – Park SG, Murray KK, 2011 J. Am. Soc. Mass om. 22(8), 1352-62. A laser on technique followed by capture of the ablated molecules by an electro-nebulization solvent jet, a technique known under the acronym of LAESI (for "Laser on oSpray Ionization") was introduced by the same team and the team of Pr. K. Murray in the same year (2007). Ablation is accomplished with a pulsed laser ng in the infrared domain. The d molecules are ionized with an electrospray jet and transferred towards the inlet of the mass spectrometer. The advantage consists in the excitation of nt biological molecules like water with a low space resolution. This technique was already used for living sms. This technique is difficult to miniaturize and requires the use of a solvent with the drawbacks which this includes within a focus for in-vivo use.
The invasive techniques will be left aside in the case of the present device and y those resorting to a bistoury regardless of its nature (manual, electrical, …).
Document US 2010/0012831 teaches an ex-vivo 3D molecular imaging method based on the LAESI technique. In the LAESI method an infrared laser is used for ablating the molecules. The ablated molecules are captured by a jet of charged organic solvent ts produced by ionization by Electrospray (ESI) and then t towards the mass analyzer via an interface using electric fields. Here, the ablated material, in order to be analyzed, should be captured by a jet of c t droplets, which is not compatible with the in-vivo use, as well as the application of an ic field. 18244134_1 (GHMatters) P105417.NZ The devices used for the LAESI technique are not compatible because of their structure for in-vivo use since they are essentially ed for o use.
Document US 7,910,881 is d to the preceding nt and has an ex-vivo analysis method based on the LADC (Laser Ablation Droplet Capture) technique. Here the sample is desorbed from the surface and then captured in a solvent placed in a capillary above the ablation point. The sample to be analyzed is ore ved in a solvent before being transferred to the mass spectrometer. A real time analysis cannot be contemplated with this device because of the time taken by the sample to arrive to the analyzer and possible material losses during the transfer. nt US 2012/0156712 proposes a system for analyzing tissues in-vivo in real time. The ablation of the tissue to be analyzed is accomplished by means of an electrode or an electric bistoury. This is therefore an invasive method.
The article "In Situ, Real-Time fication of ical Tissues by iolet and Infrared Laser Desorption Ionization Mass Spectrometry" Anal. Chem. 2011, 83, 1632-40, discusses an in situ analysis method for biological tissues within the scope of diagnostics or surgical operations of diverse types of cancers. The device consists of a laser coupled with a transfer tube which is connected to a mass spectrometer via an ionization source.
First, this ionization source has the drawback of damaging the molecules having a high molecular weight. Secondly, this device is bulky so that if it allows analysis in situ, it is not adapted to in-vivo is. Thirdly, it cannot provide the relevant information in real time.
Thus, it would be desirable to develop a device for analyzing ical material in vivo in real time based on mass spectrometry.
SUMMARY OF THE INVENTION According to the invention, a device for molecular analysis of biological material comprises: - a laser configured for emitting a wavelength comprised between 2.5 μm and 12 μm, said thereby configured laser being intended to ablate said biological material by ejecting charged and/or non-charged particles; - a mass spectrometer; and - a probe including: at least one first analysis fiber, and a transfer tube connected to the mass spectrometer, said probe being connected to the laser via said at least one first analysis fiber, said probe being connected to the mass spectrometer via said transfer tube.
Thus a urized instrument adapted for in vivo analysis is thereby available. 18244134_1 (GHMatters) P105417.NZ By analysis in the sense of the invention is meant the obtaining of lar data giving the possibility of information on the physiological condition of a patient at an instant t (diagnostic) or future instant (prognose). These molecular data may directly stem from the patient but also stem from symbiotic organisms of the t , bacterium, …).
The is fiber is connected to an ablation laser.
As an example, the wavelength of the ablation laser may be comprised between 2.8 μm and 3.2 μm. More generally, the laser is configured for providing a wavelength sed n 2.5 μm and 12 μm.
The transfer tube is connected to a mass spectrometer.
According to an additional characteristic of the device, the laser is a pulsed laser configured for generating a beam for which the energy is comprised between 2 se and 15 mJ/pulse and the surface between 30 μm2 and 3 mm2. According to an additional characteristic, a system for focussing and erring ions is interposed between said transfer tube and the mass spectrometer.
According to another additional characteristic, a metal grid, advantageously extremely thin, is introduced between the transfer tube and the mass spectrometer.
This grid advantageously gives the possibility of increasing the ivity of the analysis by increasing the production of ions.
According to another additional characteristic of the invention, a nebulization capillary is connected to the de-pressurization capillary.
Thus, the nebulization capillary is ted to a solvent distribution means.
It may be necessary to provide a second analysis fiber.
According to the invention, the device further includes a therapy fiber for laser therapy.
Advantageously, the therapy fiber is connected to a laser with a wavelength adapted for destroying cells.
The advantage of the system is to give the possibility of combining an analysis system and that of a therapy system. According to the results obtained via the analytical part, the tissues may be processed via the therapy fiber. ing to a particular embodiment, the probe r includes an illumination channel and an image shooting l.
This embodiment will quite particularly be suitable for endoscopic use.
BRIEF DESCRIPTION OF THE DRAWINGS 18244134_1 (GHMatters) P105417.NZ The present invention will now appear in a more ed way within the scope of the description which follows of an ary embodiment given as an illustration with reference to the appended figures which illustrate: - Fig. 1 shows a perspective scheme of a probe according to a f irst embodiment of the invention, - Fig. 2 shows a perspective scheme of an endoscopic probe acco rding to the invention, - Fig. 3 shows a scheme of this probe connected to the equipmen t required for its application, - Fig. 4 shows a scheme of the transfer line which connects the probe to the mass spectrometer, - Fig. 5 shows a spectrometry diagram relative to the analysis of a biological tissue ex vivo and notably of a bovine liver, more particularly: o Fig. 5a shows the total ion current over the whole of the acquisition period, o Fig. 5b, illustrates the spectrum obtained during the laser irradiation period, - Fig. 6 shows a comparison of an ex vivo analysis between the liver and the brain of a rat, more particularly, o Fig. 6a shows the s obtained on the liver, o Fig. 6b shows the s obtained on the brain, - Fig. 7 shows an analysis of a cancer biopsy of a lymphoma of a dog - Fig. 8 shows analyses of digital imprints, a comparison man/w oman (in vivo is on fingers).
DETAILED DESCRIPTION The elements present in several figures are assigned a single and same reference.
With reference to Fig. 1, a probe has been illustrated according to the invention in quasi-contact with a ical material to be analyzed P, the skin of a t in this case (but this may quite also be an organ).
The probe S appears as a cylinder in which appear an analysis fiber A and a transfer tube T. Both of these elements are flushed with the front face of the probe, the one which comes in proximity to the biological material to be analyzed. The function of these elements is explained later on. 18244134_1 (GHMatters) P105417.NZ This is the ment at the basis of the invention which gives the possibility of carrying out an al analysis on the skin, the hair, the nails or internally on an organ during open surgery.
According to a development of the ion, the probe is in fact an endoscopic probe which includes additional elements.
With reference to Fig. 2, the endoscopic probe 10 appears here as a cylinder having an axial recess 11.
It has an analysis face or a front face which is visible in the figure and which also has an opposite face, the rear face which does not appear in the figure.
The actual recess 11 is also cylindrical and a transfer tube 21 is ed therein.
The function of this transfer tube is detailed further on.
In parallel with the recess 11 are placed l elements which then also are cylindrical.
First, an illumination channel 12 such as an optical fiber opens onto the rear face so as to be connected to an illumination device not shown in this figure.
Secondly, an image shooting channel 13 is positioned in proximity to the illumination channel 12. It includes an image shooting apparatus such as a camera and the output on a rear face is accomplished through a video link 23.
Thirdly, a first optical analysis fiber 14 which is flushed with the analysis face opens onto the rear face. Its connection is explained uently.
Advantageously, a second optical analysis fiber 15 may be provided which also opens onto the rear face.
For the case when the probe is also used for ent by laser therapy, the latter further includes a laser therapy fiber 16 which again opens onto the rear face.
With reference to Fig. 3, the different connections of the probe 10 are explained.
The illumination channel 12 is ted to an illumination device 32.
The video link 23 is connected to a viewing screen 33.
The transfer tube 21 is connected to a mass spectrometer 31. The detail of this connection is provided later on.
This transfer tube preferably ts of a material aiming at minimizing the absorption phenomena in order to ensure an efficient transfer of the ablated material through the analysis fiber. This material is for example PTFE.
The first analysis fiber 14 is connected to a first ablation laser 34. This laser 34 has the function of sampling the tissue which it irradiates thereby causing ejection of charged particles (molecular ions) and/or non-charged particles in a gaseous phase. 18244134_1 ters) P105417.NZ The wavelength of this laser may be selected in the domain which extends from infrared to ultraviolet, preferentially in the infrared.
For example this is a laser with a wavelength comprised n 2.8 μm and 3.2 μm, typically an erbium-YAG laser emitting at a wavelength of 2.94 μm.
Mention may also be made of: - Neodymium-YAG lasers: 1.064 μm; 0.532 μm; 0.355 μm; 0.266 μm - Xe-Ne lasers: from 2 μm to 4 μm - HF (Hydrogen fluoride) : 2.6 μm - Fiber lasers of the Ytterbium type, doped with bismuth, thuli um or holmium: from 1.07 μm to 2.1 μm.
These lasers may be used as direct sources of emission or coupled with an OPO (Optical Parametric ator). An OPO actually allows from a laser wave of a given wavelength to produce two waves with a larger wavelength. This therefore gives the possibility of widening the range of wavelength which may be used for the relevant laser, seen by the biological al to be ablated.
Generally, it is sought to cover the range of wavelengths from 2.5 μm to 12 μm.
Indeed, in this range of wavelengths, the absorption bands of O-H, N-H, C-H, C=O, C=N, C=C, C-O, C-N and C-C bonds are covered, which may be present in the biological al to be ablated.
In particular, in this range of wavelengths from 2.5 μm to 12 μm, the laser gives the possibility of ablating the biological material by generating at least charged particles, in particular molecular ions, in a gas phase.
However it is possible to limit f to the range of wavelengths comprised between 2.5 μm and 3.5 μm. Indeed, in this range of wavelengths, the absorption bands of the O-H, N-H and C-H bonds are covered.
It is le to limit oneself to a range of more limited wavelength, comprised n 2.8 μm to 3.2 μm. Indeed, in this range of wavelengths, the absorption bands of the O-H and N-H bonds are covered.
Moreover, the on laser 34 will advantageously be a pulsed laser configured for generating a beam for which the energy is advantageously comprised between 2 mJ/pulse and 15 mJ/pulse, the surface of the beam (focussing) being sed between μm2 to 3 mm2.
The energy of the beam may be comprised between 5 mJ/pulse and 12 mJ/pulse, or further between 5 mJ/pulse and 10 mJ/pulse, the surface area of the beam (focussing) being comprised between 30 μm2 to 3 mm2. 18244134_1 (GHMatters) P105417.NZ For the ranges of energies considered earlier, the laser beam may moreover have a e area comprised between 100 μm2 and 3 mm2, between 10-3 mm2 and 3 mm2, between 10-2mm2 and 3 mm2, between 10-1mm2 and 3 mm2, between 0.5 mm2 and 3 mm2, or between 0.5 mm2 and 2 mm2. In particular, the beam of the laser may typically ablate a volume of biological material for which the base e is of the order of the 1 mm2, which substantially corresponds to a beam for which the surface area or focussing is also of the order of 1 mm2.
The ors have actually been able to ascertain that this selection in the energy provided by a pulse of the ablation laser 34 and in the surface area (focussing) of this beam ensured better production of molecular ions and therefore brought a synergistic effect with the selection of the range of ngths of the laser specified above.
Moreover it should be noted that the penetration depth of the laser beam is typically of a few microns per laser pulse.
If a second diagnostic fiber is provided, the latter is connected to a second ablation laser of a type different from the first. The possibilities of the diagnostic are thereby increased. Let us mention as an example the Neodymium-YAG laser at a wavelength of 0.532 μm. Let us also mention as an example also r laser, optionally coupled with an OPO for acting in the range from 2.5 μm to 3.5 μm. Said second analysis fiber advantageously giving the possibility of increasing the analysis ilities.
The thereby ejected particles are d by the transfer tube 21 which will forward them to the mass spectrometer 31.
The laser therapy fiber 16 is connected to a therapy laser 36. Actually, if the analysis carried out earlier reveals that the tissues have to be treated, the treatment may take place immediately, this without using any piece of additional equipment. Thus, for example it is le to use for therapy, a laser (laser diode) emitting at 980 nm, as proposed by Gonzalez-Mertinez et al., "Robot-assisted stereoactic laser ablation in medically intractable epilepsy: operating technique, Neurosurgery, 2014, suppl 2: 167- With nce to Fig. 4, a connection example is ed between the probe 10 and the mass ometer 31.
The transfer tube 21 is extended with a de-pressurization capillary 41 on the opposite side to that of the probe 10. The capillary is preferably a metal capillary. It has a smaller internal diameter than that of the transfer tube 21. This gives the possibility of sing the de-pressurization of the mass spectrometer 31, which s an acceleration of the particles. If this capillary 41 is a metal capillary, it is possible to 18244134_1 (GHMatters) P105417.NZ generate an electric field for generating a potential difference between this capillary and the inlet of the mass spectrometer 31.
The de-pressurization capillary 41 is extended at the inlet of the mass spectrometer 31 with a system for focussing and transferring ions 42 integrated inside the mass spectrometer.
Alternatively, this system 42 may be positioned outside the mass spectrometer, upstream from the , with nce to the path of charged or non-charged particles.
Moreover, also alternatively, this system 42 may be a system for focalizing ions or a system for transferring ions.
This system 42 gives the possibility of guiding the aerosol including charged particles or not towards the mass spectrometer.
The transfer tube 21 may be provided with a heating means 44 for increasing its temperature.
It is also possible to provide a nebulization ary 45 which will be connected to the de-pressurization capillary 41. This nebulization capillary 45 is supplied with a distributor of solvent 46. A l member 47 is provided for regulating the distributor 46 so that the solvent flow rate is the d one. This solvent gives the possibility of ucing a conventional electrospray process in order to increase the production yield of charged molecules (molecular ions). The advantage of introducing the solvent in this location is the e of toxicity both towards the users and towards biological tissues.
However, this is only proposed as an option. Also, it is possible to contemplate that no electrospray means connected to a t distribution means is ed n the transfer tube T, 21 and the mass spectrometer 31. , an advantage of the invention is that the ablation process of the biological material gives the possibility of ejecting at least charged particles (molecular ions) in a sufficient amount of their subsequent analysis with the mass ometer.
Moreover, it is also possible to provide a distribution capillary which will be connected to the de-pressurization capillary. This distribution capillary is then supplied by a gas butor containing GH+ ions. This gives the possibility of inducing, by collision, a transfer protons to the particles to be analyzed and thereby increase the yield of the ion production. This case is not illustrated in the appended figures, but the implantation of this distribution capillary and of the gas distributor may be similar to that of the electrospray ary 45 and to its solvent distributor 46, respectively. r, this is only proposed as an option. Indeed, it may be contemplated that no distribution capillary connected to a gas distributor is provided between the transfer tube T, 21 and the mass spectrometer 31. Indeed and as a er, an advantage of the 18244134_1 (GHMatters) P105417.NZ invention is that the ablation s of the biological material gives the possibility of ejecting at least charged particles (molecular ions) in a sufficient amount for their subsequent analysis by the mass spectrometer.
It is also possible to provide a metal grid 48 in the transfer line which goes from the probe 10 to the mass spectrometer 31.
This grid is for example positioned between the transfer tube 21 and the depressurization capillary 41, as illustrated in Fig. 4.
This is an ely thin grid of the type of those which are used in electron microscopy. Its function is to break the particles or the aggregates of particles ejected by the ablation laser 34 so that they are finally accelerated intended for the mass spectrometer 31. This grid 48 does not break the molecules, some of them are in an ionic form in the transfer tube T, 21, but larger particles.
A mass spectrometer conventionally ses and in the following order: - a source - a system for transferring and focussing ions - at least one mass analyzer It also comprises a detection system.
According to a particular embodiment of the invention, the mass spectrometer does not include any source and comprises at least one system for focussing and/or transferring ions interposed between the transfer tube and said mass analyzer.
In a non-limiting way, mention may be made as examples of system for focussing and/or erring ions, a transfer capillary, a skimmer, a focussing lens, a transfer system with multipolar , an ion funnel, an electrostatic lens.
The mass spectrometer may also e elements aiming at improving its performances such as for e an ion mobility system.
According to a possible embodiment, the system for ing and transferring ions is a transfer capillary.
The spectrometer ses a mass analyzer 49. The mass analyzer used may be of any type but should be simple (eg. Simple Magnetic Sector (B) or with double focussing (BE, EB), pole (Q), Ion trap (IT), time of flight (TOF), cyclotronic ion resonance (CIR), orbitrap), combined (eg. Triple Quadripole) or hybrid (eg. Q-orbitrap, QTOF Alternatively, the mass analyzer used may be another system for separating ions (e.g. ion mobility).
Let us now tackle the contemplated strategy upon using the present invention. 34_1 (GHMatters) P105417.NZ The device according to the invention operates on the basis of a laser ablation process giving the possibility of sampling the biological material to be analyzed. This leads to the ejection of les in a gas phase (either charged or not). The ablated material is delivered in real time to the mass spectrometer 31 via the transfer tube 21.
Indeed, the ablation process and the ejection of particles, related to this ablation, as well as the transit time in the transfer tube are short and may be described as real time. This gives the possibility of ting the molecular profiles (signals ng from the is of the biological material corresponding to biomolecules of the types of c compounds, amino acids, metabolites, lipids, peptides, …) characteristic of the analyzed area.
Advantageously, these profiles will be compared with a data bank of lar profiles ed by the use of the present device, in real time allowing information to be obtained in a rapid way.
The bank of molecular data is established by using the present device in an ex- vivo way on biopsies of patients illustrating different grades and stages of the relevant pathology. A cohort of samples from healthy patients or not is also integrated into the data bank.
However it is possible to establish the data bank in another way.
According to a particular use, the n will displace the probe at the surface or at the inside of the patient over the relevant biological material in order to ine whether it is located in a cancer area or not ng him/her to rapidly contemplate a treatment for the patient and notably the areas which he/she will have to remove surgically. These areas to be removed may advantageously be removed by the therapy fiber according to a particular embodiment of the device according to the invention.
The invention gave the possibility of ing the following results.
Result 1: Analysis of biological tissues ex vivo A laser emitting nanosecond pulses at a frequency of 10 Hz (Quantel Easy Brillant, Les Ulis, France), connected to an OPO system with a crystal of the LiNbO3 (variable in wavelength between 2.5 and 4.5 μm, LaserSpec, Malonne, Belgium) adjusted to a wavelength of 2,940 nm is used. A Teflon transfer tube (inner diameter 10 mm) is used for erring charged and non-charged particles, and is ly connected to a mass spectrometer of the ion trap type for which the source has been withdrawn (HCT Ultra, Bruker Daltonics, Bremen, Germany). The l of N2 of the mass spectrometer was disconnected in order to allow the addition of a pump aiming at increasing the suction flow 18244134_1 (GHMatters) P105417.NZ rate in the transfer tube. The analysis of the nds stemming from laser irradiation is d out with the mass spectrometer in a negative mode over a mass over charge (m/z) ratio range comprised between 150 and 1,000.
The first experiment shown in Fig. 5 is the analysis ex vivo of a piece of bovine liver. On the latter, an irradiation of 7 mJ/laser shot over an area of 1 mm2 (1 laser shot = 1 laser pulse) is achieved. A number of 3 phases was selected during the acquisition step: a first step in absence of laser irradiation, a phase with laser irradiation and another phase in the e of laser ation. Fig. 5A illustrates the total ion current over the whole of the acquisition period and Fig. 5B shows the spectrum obtained during the laser irradiation period. The total ion current shows that the presence of a detected signal is in correlation with laser irradiation. ore, there are no compounds which adhere to the internal wall of the transfer tube and this shows a rapid analysis, in real time. Typically, the analysis time is less than 1 second.
In Fig. 6, a comparison of 2 organs in rat, the brain and the liver is carried out on an acquisition on each organ. An irradiation of 30 seconds at 7 mJ/laser shot over a region of each organ. Fig. 6 shows a m/z ratio difference between these 2 organs which shows a specificity of molecular composition of the brain as compared with the rat liver. In Fig. 7, an analysis of a cancer biopsy of a lymphoma of dogs is carried out. An irradiation of 30 seconds at 7 mJ/laser shot is carried out over a region of this . The recorded spectrum is selected at the laser irradiation period. It shows a signal generation ponding to lipids and to fatty acids and a few signal ascribable to peptides. The present invention gives the possibility of carrying out analysis in real time without contamination at the wall of the Teflon tube but also between different organs stemming from a same animal. The real time analysis carried out on the different organs shows a significant number of s which may pond to fatty acids, metabolites and lipids.
With reference to Fig. 6, the present invention has the capability of detecting different profiles according to the investigated organs and to their logical status (e.g. healthy versus carcinogenic). The advantage of this analysis is a large disparity in the es of detected molecules which may add a significant value both on the data bank of molecular profiles but also on the characterization of the biological tissues.
Result 2: Analysis of biological s in vivo In Fig. 8, a real time analysis in-vivo of the tissues of the skin of individuals of different gender is carried out at the fingers. An irradiation of 10 seconds at 9 mJ/laser shot (1 laser shot = 1 laser pulse) for each individual is carried out by means of a laser ng 18244134_1 (GHMatters) P105417.NZ nanosecond pulses at a frequency of 10 Hz (Quantel Easy Brillant, Les Ulis, France), connected to an OPO system with a crystal of the LiNbO3 type (variable in ngth between 2.5 and 4.5 μm, LaserSpec, Malonne, m) adjusted to a wavelength of 2,940 nm. A Teflon transfer tube (inner diameter 10 mm) is used for transferring either charged and arged particles, and is directly connected to a mass ometer of the ion trap type for which the source has been awn (HCT Ultra, Bruker Daltonics, Bremen, Germany). The arrival of N2 of the mass spectrometer was disconnected in order to allow the addition of a pump aiming at increasing the n flow rate in the er tube. The analysis of the compounds stemming from laser irradiation is carried out with the mass spectrometer in a negative mode over a range of mass over charge ratio (m/z) comprised between 150 and 1,000. A number of 3 phases was selected during the acquisition step: a first phase in the absence of laser irradiation, a phase with laser ation and another phase in the absence of laser irradiation. A distinction of the different individuals is viewed in Fig. 8. The present invention gives the possibility of carrying out analysis in vivo in real time on duals and to be able to observe molecular profile specific according to the gender of the individual. This analysis in vivo on individuals shows the non-invasive and painless effect of the t invention during several seconds of irradiation.
Another advantage of the present invention is the non-invasive effect on organs during the actual irradiation for several tens of seconds on a same point. Based on the molecular profile, the present invention is used for differentiation of very d areas (diameter of the irradiation area of 400 μm) of biological tissues.
The invention also relates to a method for molecular analysis of biological material, characterized in that it comprises the following steps: - emitting a laser beam at a wavelength comprised between 2.5 μ m and 12 μm towards a biological material, the interaction between this beam and the biological material causing the ablation of the latter and the on of at least charged particles, notably lar ions; - directing the ablated biological material including at least said charged particles towards a mass spectrometer, via a transfer tube; and - analyzing the composition of the ablated biological material in the mass spectrometer.
This molecular analysis method of biological material may be carried out in vivo.
The laser beam may have a wavelength comprised between 2.5 μm and 3.5 μm or between 2.8 μm and 3.2 μm. 18244134_1 (GHMatters) P105417.NZ Moreover, the laser beam may be pulsed and have an energy comprised between 2 mJ/pulse and 15 mJ/pulse, the surface of the beam (focussing) being comprised between 30 μm2 and 3 mm2.
The energy of the beam may be comprised between 5mJ/pulse and 12 mJ/pulse, or further between 5 mJ/pulse and 10 se, the surface of beam (focussing) being comprised between 30 μm2 and 3 mm2.
For the ranges of energies considered earlier, the laser beam may moreover have a surface area comprised between 100 μm2 and 3 mm2, between 10-3 mm2 and 3 mm2, between 10-2mm2 and 3 mm2, between 10-1mm2 and 3 mm2, between 0.5 mm2 and 3 mm2, or between 0.5 mm2 and 2 mm2. In particular, the beam of the laser typically gives the possibility of ablating a volume of biological material for which the base e is of the order of the 1 mm2, which substantially corresponds to a beam also of the order of 1 mm2.
Further, this method may e a step for heating the ablated biological al.
This method may also provide a step during which the ablated biological material is sifted with a grid, for example a metal grid.
Finally, it should be noted that the invention also proposes the use of a lar analysis device according to the invention for ablating said ical material by ejecting charged and/or non-charged particles, and more specially for ablating said biological material by ejecting at least charged particles, y molecular ions.
In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the word "comprise" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e. to specify the presence of the stated features but not to preclude the presence or addition of r es in various embodiments of the invention.
It is to be tood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art in any country. 18244134_1 (GHMatters) P105417.NZ
Claims (22)
1. ) A molecular analysis device for a biological material comprising: 5 - a laser configured for emitting a wavelength sed betwee n 2.5 μm and 12 μm, said thereby configured laser being intended to ablate said biological material by ng charged and/or non-charged particles; - a mass spectrometer; and - a probe including: 10 o at least one first analysis fiber, and o a transfer tube, said probe being connected to the laser via said at least one first analysis fiber, said probe being connected to the mass spectrometer via said transfer tube. 15
2. ) The device according to claim 1, wherein the laser is configured for emitting a wavelength comprised between 2.5 μm and 3.5 μm.
3. ) The device according to any one of the preceding claims, wherein the laser is configured for emitting a wavelength comprised between 2.8 μm and 3.2 μm.
4. ) The device according to any one of the preceding claims, n the laser is a pulsed laser configured for generating a beam for which the energy is comprised between 2 mJ/pulse and 15 mJ/pulse and the surface of which is sed between 30 μm2 and 3 mm2.
5. ) The device according to any one of the preceding claims, n the laser is a pulsed laser configured for generating a beam for which the energy is comprised between 5 mJ/pulse and 12 mJ/pulse, for example n 5 mJ/pulse and 10 mJ/pulse and the surface is comprised between 30 μm2 and 3 mm2.
6. ) The device according to one of claims 4 or 5, wherein the laser is a configured for generating a beam for which the surface area is comprised between 0.5 mm2and 2 mm2. 34_1 (GHMatters) P105417.NZ
7. ) The device according to any one of the preceding claims, wherein a system for focussing and/or transferring, molecular ions intended to be formed during ablation of biological material, is interposed between said transfer tube and the mass spectrometer. 5
8. ) The device according to claim 7, wherein said focussing and/or transfer system is a transfer capillary.
9. ) The device ing to any one of the preceding claims, wherein said transfer tube is provided with a g means.
10. ) The device according to any one of claims 6 to 8, wherein a depressurization capillary is inserted between said transfer tube and said focussing and/or transfer system, the inner diameter of this de-pressurization capillary being less than that of the transfer tube.
11. ) The device according to claim 10, wherein the de-pressurization capillary is a metal capillary and in that a means is provided for generating a potential difference between said de-pressurization capillary and the mass spectrometer. 20
12. ) The device according to any one of the preceding claims, wherein a metal grid is introduced between said transfer tube and said mass ometer.
13. ) The device according to any one of claims 10 to 12, wherein an electrospray ary is ted to said de-pressurization capillary and further 25 connected to a means for distributing t.
14. ) The device according to any one of claims 1 to 12, wherein no electrospray capillary ted to a means for distributing solvent is provided n the transfer tube and the mass spectrometer.
15. ) The device according to any one of claims 1 to 13, wherein a distribution capillary is connected to said de-pressurization capillary and further connected to a gas distributor. 18244134_1 (GHMatters) P105417.NZ
16. ) The device according to any one of claims 1 to 14, wherein no bution capillary connected to a gas butor is provided between the er tube and the mass spectrometer. 5
17. ) The device according to any one of the preceding claims, wherein said probe includes a second analysis fiber.
18. ) The device according to any one of the preceding claims, wherein said probe further includes a therapy fiber.
19. ) The device according to claim 18, wherein said therapy fiber is connected to a therapy laser.
20. ) The device according to any one of the preceding claims, wherein said 15 probe further includes an illumination channel and an image shooting channel.
21. ) The device according to any one of the preceding claims, characterized in that the laser is assisted with an optical parametric oscillator (OPO). 20
22. ) A molecular analysis device as claimed in claim 1 and substantially as herein bed with reference to the drawings. 34_1 (GHMatters) P105417.NZ WO 46748 WO 46748 II. 33 10 . .. . .1
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR1458925 | 2014-09-22 | ||
FR1458925A FR3026189B1 (en) | 2014-09-22 | 2014-09-22 | REAL TIME IN VIVO MOLECULAR ANALYSIS DEVICE |
PCT/IB2015/057301 WO2016046748A1 (en) | 2014-09-22 | 2015-09-22 | Device for real-time in vivo molecular analysis |
Publications (2)
Publication Number | Publication Date |
---|---|
NZ730251A NZ730251A (en) | 2022-03-25 |
NZ730251B2 true NZ730251B2 (en) | 2022-06-28 |
Family
ID=
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2015323373B2 (en) | Device for real-time in vivo molecular analysis | |
JP6594288B2 (en) | System and method for identification of biological tissue | |
KR101934663B1 (en) | An inlet instrument device for an ion analyzer coupled to a rapid evaporation ionization mass spectrometry (" REIMS ") device | |
JP6753862B2 (en) | Improved ionization of gas samples | |
JP6858705B2 (en) | Collision surface for improved ionization | |
CN107645938B (en) | Image-guided ambient ionization mass spectrometry | |
CN110312486B (en) | Laser apparatus and tissue characterization method | |
CN107636794A (en) | Liquid trap or separator for electrical surgical appliance | |
EP3265819A1 (en) | Chemically guided ambient ionisation mass spectrometry | |
CN107635477A (en) | For being coupled to rapid evaporation MALDI-MS(“REIMS”)The entrance instrument of the ion analyser of device | |
KR20170129179A (en) | In vivo endoscopic tissue identification tool | |
Ye et al. | A vision for better health: mass spectrometry imaging for clinical diagnostics | |
US20210033623A1 (en) | Tissue analysis by mass spectrometry | |
Simon et al. | Sample preparation free tissue imaging using laser desorption–rapid evaporative ionisation mass spectrometry (LD-REIMS) | |
NZ730251B2 (en) | Device for real-time in vivo molecular analysis | |
US11583182B2 (en) | Method for multimodal tissue imaging based on resonance Raman effect on metal based MRI contrast agents and method for ionizing laser plumes through atmospheric pressure chemical ionization | |
Simon et al. | Enabling Cellular Resolution Molecular Pathology for Surgical Interventions Using Laser Desorption–Rapid Evaporative Ionization Mass spectrometry | |
Ogrinc et al. | Ambient Mass Spectrometry Imaging by Water-Assisted Laser Desorption/Ionization for Ex Vivo and in Vivo Applications | |
KR20190052844A (en) | A method for treating biological tissue, a laser processing apparatus, and an atmospheric pressure mass spectrometry imaging system |