[go: up one dir, main page]

WO2003082117A1 - Dispositif et procede de quantification de corps par des ultrasons - Google Patents

Dispositif et procede de quantification de corps par des ultrasons Download PDF

Info

Publication number
WO2003082117A1
WO2003082117A1 PCT/EP2003/003227 EP0303227W WO03082117A1 WO 2003082117 A1 WO2003082117 A1 WO 2003082117A1 EP 0303227 W EP0303227 W EP 0303227W WO 03082117 A1 WO03082117 A1 WO 03082117A1
Authority
WO
WIPO (PCT)
Prior art keywords
bubbles
bodies
ultrasound
signals
transducer
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.)
Ceased
Application number
PCT/EP2003/003227
Other languages
German (de)
English (en)
Inventor
Michael Reinhardt
Peter Hauff
Andreas Briel
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Bayer Pharma AG
Original Assignee
Schering AG
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Schering AG filed Critical Schering AG
Priority to EP03745194A priority Critical patent/EP1487345A1/fr
Priority to AU2003232191A priority patent/AU2003232191A1/en
Priority to JP2003579664A priority patent/JP2005527270A/ja
Publication of WO2003082117A1 publication Critical patent/WO2003082117A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B8/00—Diagnosis using ultrasonic, sonic or infrasonic waves
    • A61B8/48—Diagnostic techniques
    • A61B8/481—Diagnostic techniques involving the use of contrast agents, e.g. microbubbles introduced into the bloodstream
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B8/00—Diagnosis using ultrasonic, sonic or infrasonic waves
    • A61B8/48—Diagnostic techniques
    • A61B8/483—Diagnostic techniques involving the acquisition of a 3D volume of data
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6891—Pre-targeting systems involving an antibody for targeting specific cells
    • A61K47/6897—Pre-targeting systems with two or three steps using antibody conjugates; Ligand-antiligand therapies
    • A61K47/6898—Pre-targeting systems with two or three steps using antibody conjugates; Ligand-antiligand therapies using avidin- or biotin-conjugated antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
    • A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
    • A61K49/223—Microbubbles, hollow microspheres, free gas bubbles, gas microspheres
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01S—RADIO DIRECTION-FINDING; RADIO NAVIGATION; DETERMINING DISTANCE OR VELOCITY BY USE OF RADIO WAVES; LOCATING OR PRESENCE-DETECTING BY USE OF THE REFLECTION OR RERADIATION OF RADIO WAVES; ANALOGOUS ARRANGEMENTS USING OTHER WAVES
    • G01S7/00—Details of systems according to groups G01S13/00, G01S15/00, G01S17/00
    • G01S7/52—Details of systems according to groups G01S13/00, G01S15/00, G01S17/00 of systems according to group G01S15/00
    • G01S7/52017—Details of systems according to groups G01S13/00, G01S15/00, G01S17/00 of systems according to group G01S15/00 particularly adapted to short-range imaging
    • G01S7/52023—Details of receivers
    • G01S7/52036—Details of receivers using analysis of echo signal for target characterisation
    • G01S7/52038—Details of receivers using analysis of echo signal for target characterisation involving non-linear properties of the propagation medium or of the reflective target
    • G01S7/52041—Details of receivers using analysis of echo signal for target characterisation involving non-linear properties of the propagation medium or of the reflective target detecting modification of a contrast enhancer, e.g. detecting the destruction of a contrast agent by an acoustic wave, e.g. loss of correlation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B8/00—Diagnosis using ultrasonic, sonic or infrasonic waves
    • A61B8/42—Details of probe positioning or probe attachment to the patient
    • A61B8/4209—Details of probe positioning or probe attachment to the patient by using holders, e.g. positioning frames
    • A61B8/4218—Details of probe positioning or probe attachment to the patient by using holders, e.g. positioning frames characterised by articulated arms

Definitions

  • the invention relates to a device and a method for the quantification of bodies contained in an examination object, which can be excited to independent, characteristic signals by diagnostic ultrasound, in particular those signals which are associated with the destruction of a body, preferably of bubbles. Furthermore, the invention also relates to a use of the device for ultrasound diagnosis and / or therapy on humans or animals and a use of the device and a method for the in-vivo and ex-vivo mapping of physiologically high or low-regulable molecular markers in Organs and tissues from postembryonic ontogenesis and from pathologically high or down-adjustable molecular markers in organs and tissues during pathogenesis and for the characterization of cell cultures by in-mapping of up and down-adjustable molecular markers of cells.
  • Ultrasound diagnostics has found a permanent place in everyday clinical practice. Details can be shown better in ultrasound recordings by injection of contrast media, since stronger ultrasound signals can be generated in this way.
  • All very small bodies (micro-bodies) that deliver a detectable signal that can be distinguished from the tissue signal, in particular gas-filled very small (micro) bubbles, are suitable as contrast agents. These bodies are fed to the examination object and can be destroyed in the ultrasound field, for example in the case of bubbles, under certain conditions. These bubbles emit independent, characteristic ultrasound signals. If the bubbles are destroyed by the ultrasound field, the independent, characteristic signals are referred to as "stimulated acoustic emission (SAE)".
  • SAE stimulated acoustic emission
  • Bubbles that can emit independent characteristic signals are described, for example, in EP 0 398 935 B1, EP 0 644 777 B1, EP 0 458 745 A1 and WO 01/68150 A1.
  • SAE stimulated acoustic emission
  • FIG. 2 an ultrasound sectional image from which the size of the SAE signals can be seen
  • FIG. 3 a conventionally recorded ultrasound sectional image.
  • FIG. 1 shows a first sound wave with low amplitude 1 and a second sound wave with high amplitude 2.
  • a bladder 3 that is sonicated remains intact. A scatter signal is generated.
  • the bladder 4 is destroyed when a sound wave with high amplitude is irradiated, a SAE signal being generated.
  • SAE signals of various bubbles can be detected according to US 5,425,366 A, US 6,186,951 B1 and DE 198 13 174 A1 using color Doppler mode.
  • SAE signals can also be displayed independently of the movement of the bubbles with other ultrasound device modes, which were originally developed primarily to represent movement.
  • the spectral Doppler, power Doppler, color Doppler and harmony method (2nd, sub, wideband, pulse inversion, ultraharmonic, color Doppler, harmonic power Doppler method) are mentioned as examples.
  • the SAE signal strength is so high that even individual bubbles can be detected.
  • the individual SAE signal is shown on the monitor considerably larger (diameter approx. 0.5 - 1 mm depending on the device, transducer and settings) than it corresponds to the actual size of the undestroyed bladder ( ⁇ 10 ⁇ m). 2 shows the size of SAE signals in the sectional view.
  • the layer thickness of a single image is approximately 1 mm, depending on the device setting. This is illustrated in Figure 3.
  • the layer thickness of a conventional sectional image can be seen there.
  • a bladder concentration of approx. 2,000-3,000 blisters / ml tissue / blood leads to a complete SAE contrast in the sonographic cross-section.
  • the saturation bubble concentration is determined by the layer thickness of the ultrasound sectional image (shown by way of example in FIG. 3) and by the size of an SAE signal shown in the ultrasonic sectional image when a single bubble is destroyed (several individual bubbles are shown in FIG. 2).
  • a low saturation concentration / saturation bladder concentration can be advantageous for qualitative imaging, since in particular, even at low bladder concentrations, a complete contrast is created in the sectional image.
  • a low saturation concentration / saturation bubble concentration is disadvantageous for the quantification of bodies, since even small concentrations of bodies, in particular bubbles, lead to contrast saturation in the image and their concentrations above the saturation concentration in the image are no longer absolute determine or no longer distinguish bodies, especially bubbles.
  • the saturation concentration / saturation bladder concentration is usually exceeded because the enriching tissue or the enriching structures are so dense, that the bodies accumulate there in a concentration above the saturation concentration / saturation bubble concentration.
  • the saturation concentration / saturation bladder concentration in the target area could also be undercut here, but the enriched number of bodies, in particular bubbles, would be in an unknown ratio to the maximum saturation. Such a measurement would therefore neither be reproducible nor would it provide a value that correlates with the number of enrichment factors present there. Therefore, a method for determining the number of bodies (number of bubbles) above the saturation concentration / saturation bubble concentration is desirable.
  • Bodies in particular bladders, can accumulate non-specifically or specifically on or in cells, tissues, organs or other structures, for example.
  • This enrichment mechanism is often referred to in the literature as passive or active targeting (Lanza, G., Wickline, S .: “Targeted ultrasonic contrast agents for molecular imaging and therapy”.
  • Dayton, PA, Ferrara, KW “Targeted imaging using ultrasound”. J. Magn. Reson. Imaging, 2002, 16: 362-377).
  • phagocytosis of bodies / bladders in organs of the reticuloendothe- lial system (RES) or in other cells which e.g. disease-related, such as in tumor-associated macrophages or macrophages in atherosclerotic processes, or tumor cells themselves or cells that are stimulated to phagocytosis, e.g. Endothelial cells of the RES.
  • RES reticuloendothe- lial system
  • Liver with certain diseases of the liver b) by adhesion or by slowly rolling bodies / blisters along, for example on the vascular endothelium; c) by immigration of bodies / blisters in or by fenestrations, eg of the endothelium, in "vascular cul-de-sacs" or "blind” blood vessels; this can occur in the transition area to necrosis, in the case of severely edematous or inflammatory processes, hemorrhages, hematomas or bulges of the vascular system, such as aneurysms, vascular tumors, such as hemangiomas, or by washing into necrosis; d) by transporting phagocytized or adherent bodies / bladders, which are phagocytosed, for example, by monocytes or adhered to cells of the blood, such as leukocytes, through endothelial barriers, such as the blood-brain barrier, to pathological areas, for example.
  • a specific enrichment of bodies, in particular bubbles can be achieved by a covalent or non-covalent, including an electrostatic bond, hydrogen bond or Van der Waais bond, to physiological or pathophysiological structures.
  • the bodies, in particular bubbles carry target-specific binding molecules on their surface in these cases.
  • 3D-Echotech, Halbergmoos, Germany has also attempted to satisfy the need for the quantifiability of bubbles with an ultrasound device for the quantification of bubble signals in the blood.
  • the video image of the ultrasound device is digitized, broken down into color and gray values and optionally evaluated according to the average brightness value or the number of screen pixels within a defined measurement window (ROI).
  • ROI measurement window
  • the field of application of this measuring system is to quantify the blistering in blood vessels based on the brightness values within a definable ROI.
  • this method is also unsuitable for quantifying the bubbles above a concentration of approx. 3,000 particles or bubbles / ml. Even below this concentration, a reproducible measurement is not possible with this device.
  • a disadvantage of the known methods and devices is that it is not possible to display small structures, since the sound lobes emanating from a transducer have minimum dimensions that limit the resolution. For example, the thickness of the measuring layer detected by the sound waves is approximately 1 mm, so that finer structures can no longer be resolved.
  • the cell cultures are generally immortalized cells (which can be expanded in vitro indefinitely by special manipulation) and cannot reflect the real Vo ratios.
  • Another very common method of identifying molecular markers is by immunohistochemical examination of tissues. Tissues of special interest, for example tumors, are histologically processed after their removal and immunohistologically specifically examined for the expression of one or more molecular markers. Only after histological identification of such markers can they be imaged later in vivo by using appropriate antibodies which are coupled to corresponding signal generators.
  • the present invention is therefore based on the object of finding a method and a device for the quantification of bodies in an examination object, with which the concentration of the bodies can be determined in vivo, ex vivo and in vitro above the saturation bubble concentration is possible.
  • the drive and the device should be able to reproducibly quantify body quantities and determine their concentration.
  • the object is achieved by the device for quantifying bodies contained in an examination object according to claim 1, their use for ultrasound diagnosis and / or ultrasound therapy on humans or animals according to claim 10, their use for in-vivo and ex-v / Vo mapping of molecular markers in organs and tissues and in w ' fro mapping of cells for the characterization of cell cultures according to claim 11 and the method for quantifying bubbles contained in a test object according to claim 12 solved.
  • Preferred embodiments of the invention are specified in the subclaims.
  • Fig. 7 Multi-element transducer with defined overlapping sound fields (2D array)
  • a: SAE layer thickness 10 ⁇ m to 100 ⁇ m
  • Sectional image 15 displacement distance 100 ⁇ m
  • Fig. 10 Dependence of the MU CO rr. on the SAE layer thickness (agar phantom, example 1)
  • Fig. 11 Dependence of the SAE CO rr. on the SAE layer thickness (agar phantom, example 1)
  • Bodies in the sense of the present application are solid particles and / or bubbles with a diameter ⁇ 10 ⁇ m.
  • Solid particles contain at least one surface-active substance (e.g. surfactant), at least one carbohydrate, protein, lipid and / or natural and / or synthetic polymer.
  • Bubbles in the sense of the present application are gas-containing phases with a diameter ⁇ 10 ⁇ m, which are either free or coated.
  • the phases can be substantially spherical.
  • the envelope can contain at least one surface-active substance (eg surfactant), at least one protein, lipid and / or natural and / or synthetic polymer.
  • Bubbles according to the present application can also, for example, gas-filled microcapsules, gas-filled microparticles, gas-filled microspheres, microballoons and surfactant-stabilized microbubbles.
  • Ultrasound contrast media are agents that contain bodies, in particular bladders, for use in ultrasound diagnostics and / or therapy.
  • An independent, characteristic signal is an echo signal of a body, in particular a bladder, which is independent of the scatter signal.
  • This also includes all non-linear signals such as 2nd harmonic, subharmonic or ultra-harmonic, as well as in particular SAE signals, such as those which arise when bubbles are destroyed in the ultrasound field.
  • Bubbles preferred according to the invention can be destroyed under certain conditions when excited with ultrasound. This creates an independent signal that deviates from the transmission signal and is independent of the scattering and movement of the bubbles. This independent, characteristic signal is called stimulated acoustic emission (SAE) (synonym: Loss of Correlation (LOC)).
  • SAE stimulated acoustic emission
  • LOC Loss of Correlation
  • the saturation concentration / saturation bubble concentration is the concentration of bodies / bubbles per milliliter of object under examination, e.g. tissue or blood, above which the independent, characteristic signals, in particular the SAE signals, are superimposed within a sound field and above which one It is impossible to quantify the bubbles because the number of bodies / bubbles can no longer be inferred from the sum of the CS signals, even when using arithmetic correction formulas.
  • CS / SAE layer thickness is the concentration of bodies / bubbles per milliliter of object under examination, e.g. tissue or blood, above which the independent, characteristic signals, in particular the SAE signals, are superimposed within a sound field and above which one It is impossible to quantify the bubbles because the number of bodies / bubbles can no longer be inferred from the sum of the CS signals, even when using arithmetic correction formulas.
  • the CS / SAE layer thickness is equal to the displacement distance per ultrasound sectional image. This layer thickness is preferably significantly less than the cut layer.
  • the cut layer is created in the examination object by irradiation of a sound lobe (sound field).
  • piezo crystal elements emit sound pulses which are coupled and focused into the medium to be sonicated by suitable means.
  • a cut layer can be gradually moved away from the sound lobe and the corresponding image can be generated by the sound signals arriving at the receiver.
  • the layer thickness is defined by the feed between two images.
  • Morphometric units are a measure of the area, corresponding to a section layer, expressed in the number of screen pixels that the CS signals represented in the video image occupy.
  • the screen pixels are usually counted automatically by video densitometry. If the number of screen pixels of an individual CS signal is known, the number of bodies / number of bubbles can be concluded from the morphometric units.
  • Corrected SAE SAE CO rr.
  • corrected MU MU corr .
  • CS signals can be superimposed in the video image.
  • the probability of such a superimposition increases with increasing concentration of bodies / bubbles and layer thickness of the ultrasound sectional image, because in this case a larger number of bodies / bubbles is detected. Both parameters have a direct influence on the percentage color area of the CS signals in the video image. If the bodies / bubbles are randomly distributed in the examined volume, the value of MU can be corrected if CS signals are superimposed as long as there is no complete color saturation in the ROI:
  • FP color screen pixels that lie within the measurement window (ROI) in the area of the CS effects shown
  • MU FP within the measurement window (ROI)
  • GP Screen gray pixels that are within the measurement window (ROI) but outside the CS effects shown.
  • the corrected MU (MU cor r.) Can be converted into a the number of CS signals (SAE cor r.) are converted:
  • a 2D array consists of a one-dimensional row of piezo elements.
  • a 3D array consists of several adjacent rows of piezo elements.
  • a transducer 23 is shown schematically. The directions in relation to the transducer are defined as follows:
  • x transverse to the transducer (reference number 24: 2D array)
  • y lateral (reference number 25)
  • z axial (direction of sound propagation) (reference number 26)
  • Intra-, inter- and / or extracellular molecular structures for example receptors, ligands, enzymes or their structural components.
  • the device according to the invention and the method according to the invention serve to quantify bodies, preferably bubbles, contained in an examination object.
  • the device comprises at least one transducer for emitting an ultrasound field, which excites the bodies contained in a layer in the examination object, preferably bubbles, for emitting characteristic ultrasound signals. At least one of the transducers is also used to receive the characteristic signals.
  • at least one data processing system for determining the number of bodies, preferably bubbles is provided in the layer from the characteristic signals.
  • the ultrasound field after excitation of the bodies preferably bubbles
  • the ultrasound field after excitation of the bodies can be displaced in a first layer in the examination object such that bodies / bubbles can be excited in a second layer after the displacement, the first layer and the overlap second layer and are shifted parallel and defined against each other.
  • Ultrasound signal data records are thus obtained from different cut layers in the examination object, which contain information about the presence of bodies / bubbles in the individual cut layers, in particular about the spatial arrangement of the bubbles in the cut layers.
  • the layers are preferably shifted against one another in such a way that they overlap in the transverse direction (x direction) to the rows of piezocrystals.
  • the transducer can either be designed to emit ultrasound signals as well as to receive the signals originating from the bodies, preferably bubbles. Alternatively, the transducer can also be designed exclusively for emitting the ultrasound signals. In this case, a further receiving head is required to detect the incoming ultrasound signals from the bodies, preferably bubbles.
  • bodies / bubbles contained in a layer are excited with an ultrasound field to produce characteristic ultrasound signals.
  • the characteristic ultrasound signals emitted by the bodies / bubbles are received, and the number of bodies / bubbles is determined from the received ultrasound signals, in each case by the Displacement of the ultrasound field after excitation of bodies / bubbles in a first layer in the examination object bodies / bubbles in a second layer overlapping the first layer and excited in a defined and defined manner against this shifted layer.
  • the device according to the invention and the method according to the invention are not limited to embodiments using imaging or the video image.
  • the CS signals within a region of interest (ROI) can also already be counted on the basis of the received signals (radio frequency (RF) data), although the concentration of saturation bubbles when the CS signals are counted at the level of the received signals from the Can distinguish saturation bubble concentration when counting the CS signals in the two-dimensional ultrasound image.
  • the device according to the invention and the counting of the received signals can be integrated into an ultrasound device.
  • the device and the method according to the invention determine concentrations of bodies, preferably bubbles, which are above a limit which is in the range from 1000 to 3000 bodies (bubbles Vml:
  • the spatial resolution in the x direction or x and y direction (in a plane perpendicular to the sound lobe) when using a single-crystal ultrasound transducer with a round cross section is in the mm range in these cases.
  • the thickness of the Sound field at the narrowest point in the sound beam emitted by the transducer in the mm range is approximately 1 mm 3 . If a body, preferably a bubble, in this volume is destroyed by ultrasound, the exact spatial position of the corresponding CS signal within this volume cannot be determined exactly. Consequently, this signal is shown in the image in a size that is also in the millimeter range.
  • the present invention enables a significantly higher spatial resolution than with known methods and devices.
  • calculable volumes are used. This makes it possible for the first time to quantify bodies, preferably bubbles, in an examination volume even if the saturation concentration / saturation bubble concentration would normally be exceeded.
  • the Kretztechnik device is also unsuitable for quantification, since the overlap of the sound fields in the axial direction (z) is not uniform. In the close range, several adjacent sound fields overlap almost completely, while gaps arise with increasing distance from the transducer, which are then interpolated. As a result, the signal yield in an examined tissue also depends on the distance to the transducer. The aforementioned object is only achieved by the present invention.
  • the principle of the method according to the invention can also be used for such bodies, preferably bubbles, which can be excited with diagnostic ultrasound to form an independent and characteristic signal independent of destruction.
  • a signal can be, for example, a harmonic or subharmonic signal.
  • no independent, characteristic signals of solid particles can be detected after excitation; This can be achieved, for example, by starting the scan at a position that is free of particles, or by moving the transducer out of an area in which independent, characteristic signals of solid particles can be detected after excitation, before the scan begins. is moved to an adjacent area, in which this is not the case, b) in the subsequent images / layers, the solid particles become like this stimulated that they produce an independent, characteristic signal, c) as long as the cumulative scan distance is less than or equal to the layer thickness of the sound field, the newly added signals are determined in each image / layer.
  • the determination of the newly added signals can be determined by subtracting the respective pre-value or by simple, preferably automatic counting of the signals after sending a destruction pulse.
  • the number of signals in the overlap area can generally be determined by simple, preferably automatic, counting, as in the case of SAE signals.
  • the sectional images overlap by 99%.
  • the second sectional image only the newly added bodies, preferably bubbles, that are in the 10 ⁇ m layer are detected.
  • the distance of the shift between two images is at least 5 ⁇ m, preferably at least 10 ⁇ m and particularly preferably at least 20 ⁇ m. In a very particularly preferred embodiment, the displacement distance is at least 50 ⁇ m.
  • the shift distance is smaller than the scan thickness (approx. 1 - 2 mm).
  • the optimal distance shift depends on the highest body concentration / ml, preferably bladder concentration / ml in the scanned volume. It is advantageous to set an overlap of the volumes between 20% and 99.99%, preferably between 40% and 99.9% and particularly preferably between 70% and 99%. If the displacement distance is chosen too large or the ultrasound sectional images do not overlap at all (FIG. 5a), absolute quantification is also not possible with the method according to the invention, since there is an uncorrectable superposition of the CS signals in the ultrasonic sectional image.
  • the displacement distance can be chosen to be too large, for example, or the ultrasound sectional images may not overlap if the degree of accumulation of the body, preferably bubbles, is underestimated. In these cases, the displacement distance per ultrasound sectional image must be reduced in order to ensure absolute quantifiability.
  • 9 to 11 show the relationship between the displacement distance and the morphometric units (MU) or the number of independent, characteristic signals in the sectional image.
  • the concentration of bodies, preferably bubbles, is preferably determined at those CS / SAE layer thicknesses whose sectional images each have 20 to 80%, preferably 30 to 70%, color saturation. In favor of a reduced examination time, however, the displacement distance can be increased as long as the saturation bubble concentration is not reached in any sectional image.
  • a defined program that varies the CS / SAE layer thickness can ensure an optimal compromise between the examination time and the accuracy of the measurement.
  • the optimal displacement distance controlled by the program can be determined, for example, on the basis of a sufficiently large population of patients (examination objects).
  • the standardized scan process with a defined parallel shift of the sound fields and the higher spatial resolution of the method according to the invention enable a reproduced measurement.
  • Such a measurement can be sufficient, for example, for a comparison between different patients or for a therapy course check for the same patient. If not all bubbles are destroyed in a sectional view, you can also send one or more further destruction pulses before moving to the next position.
  • the overlap and displacement of the volumes of the cut layers can be achieved by displacing the transducer and the examination object against one another (relative to one another).
  • the defined parallel shift can be achieved by
  • the tissue or the examination object for example a patient, is shifted relative to a fixed transducer or
  • the transducer is moved relative to the fixed examination object to be examined, for example a tissue, or III.
  • a transducer is used, which is able to create overlapping and shifted sectional images one after the other, i.e. the volumes are shifted and overlapped with a transducer, which itself takes overlapping and parallel shifted ultrasound images;
  • the transducer can be controlled in such a way that overlapping and in particular transversely shifted ultrasound sectional images or sound lobes
  • Cut layer are included.
  • the examination object (patient, animal, an eccentric or isolated organ or a cell culture dish) is automatically moved past the fixed transducer (e.g. driven by a servo motor or DC motor).
  • the fixed transducer e.g. driven by a servo motor or DC motor.
  • the transducer is automatically moved over / opposite the examination object (patient, animal, an eccentric or isolated organ or a cell culture dish) with the help of a special device.
  • the device can be a frame within which the transducer is fixed in a holder and is moved by a motor drive along a parallel guide.
  • a single crystal can be shifted in the x and y directions with defined overlap areas.
  • This variant offers e.g. for culture dishes of cells on the surface of which bodies, in particular bladders, have accumulated specifically or inside which have become nonspecific, for example by phagocytosis. Here it is sufficient to state how many bodies, in particular bubbles, per area or volume have accumulated. The data does not have to be converted into pictures.
  • adjacent sound lobes can be brought to overlap within a line in the transverse direction in a defined manner with a transducer containing a 2D array.
  • the transducer can also contain a 2D array, which is moved within the transducer in parallel with a defined overlap in the transverse direction (x direction).
  • the lateral resolution can be increased within a 2D sectional image if the individual scan lines from which the sectional image is constructed also overlap in a defined manner.
  • the individual sound fields which are each generated by a group of neighboring piezo elements, are overlapped more strongly and above all by selecting small increments (down to a piezo element).
  • a sound wave in the form of a sound lobe 9 is emitted into the examination object.
  • Undestroyed bubbles 8 located within the sound lobe are destroyed and thereby emit independent, characteristic signals 9.
  • the destroyed bubbles are identified by the reference number 10.
  • a newly formed group of piezo elements of the array is excited to vibrate, which differs from the group used in the first excitation by only one piezo element by which the group is now offset (FIG. 7b). Due to the overlap of the resulting sound lobe with the previously emitted sound lobe, only the newly detected bubbles are excited into independent, characteristic signals.
  • the resolution increases by the factor in which the displacement is related to the width of the piezo element group.
  • At least one motor drive is therefore provided for displacing the examination object or the at least one transducer relative to one another.
  • the transducer If the transducer is to be moved relative to the examination object, it must be moved in a defined way.
  • a movable holder for the transducer is provided.
  • the examination object is fixed in a holding device.
  • the object is held practically motionless by the holding device.
  • a patient can be placed on a couch on which the patient rests during the examination.
  • Such holding devices can be used for holding the examination object both when it is not moved during the examination, but only the transducer, and when the examination object is moved and the transducer is not.
  • the above-mentioned holding devices can also be used for the examination object in the event that both the transducer and the examination object are moved against one another at the same time.
  • an image evaluation system for data processing and image display is provided in a preferred embodiment, with which the quantification can be carried out by video densitometry.
  • the data sets formed from the ultrasound signal sets can be reshaped in a known manner in such a way that a video image of the individual cut layers is created, each cut layer containing the information from the areas which do not overlap with a previously excited cut layer (new cut layer area (CS / SAE-layer)).
  • the bodies represented in the individual video images, in particular bubbles can then be counted manually or obtained by video-densitometric determination of the color image points (color pixels) and division of the value obtained therefrom by the average number of color pixels per bubble representation.
  • FP are the color pixels representing the body, in particular bubble signals, in a locally resolved screen representation within the overlap layer and GP are screen gray pixels in a locally resolved screen representation, which are not the pixels representing the body, in particular bubbles.
  • the device according to the invention is particularly suitable for ultrasound diagnosis and / or therapy on humans or animals. Not only in the areas of clinical diagnostics, but also in basic medical and biological research and pre-clinical research and development, new possibilities of investigation open up:
  • the bodies preferably bubbles, can be tested in vivo in animal experiments, ex vivo in harvested organs or tissues and in vitro e.g. be quantified in cell cultures in order to clarify pathological or physiological processes at the molecular level.
  • the method according to the invention can be used to test which binding systems work at all, which binding molecules are present under which conditions at which locations and in what quantity, which influencing variables determine the binding capacity and accumulation, what kinetics the body, preferably bubbles, or , in the case of ultrasound-induced active ingredient release, what amounts of an encapsulated active ingredient are released.
  • the device according to the invention and the method according to the invention can be used, for example, for the / nv / Vo mapping of physiologically high or low regulatable molecular markers in organs and tissues from postembryonic ontogenesis (ontogenesis: total development of an individual from the unfertilized / fertilized egg cell to to natural death: this comprises four phases: 1. embryonic development, 2. postembryonic development, 3. time of sexual maturity and reproduction and 4. time of aging).
  • both passively and actively accumulative bodies preferably bubbles
  • the bodies can be provided with binding molecules specific for the markers that are located on the surface (actively accumulable bodies / bubbles).
  • passively accumulative bodies can also be used, i.e. Bodies / bubbles without such specific binding molecules.
  • molecular markers can be localized in vivo on different cells in humans and animals and their influence on ontogenesis can be determined. Furthermore, a time-controlled mapping of known and newly identified molecular markers in the whole organism can be carried out in vivo. This enables a previously unknown function of the molecular marker in ontogenesis to be determined in vivo.
  • the spatially high-resolution sonography according to the invention can be used to investigate the signs of aging from which these molecular markers originate and which they can lead to, furthermore what basis these receptors represent for age-related changes or diseases of the whole organism and whether there are any findings for development be derived from medications that alleviate age-related complaints or even help to avoid them.
  • a / n-wVo mapping of pathologically high or low regulatable molecular markers in organs and tissues during pathogenesis can also be carried out using passively and actively accumulative bodies, in particular bladders ,
  • disease-associated molecular markers can be localized in vivo on different cells in humans and animals and therefore their influence on the pathogenesis of diseases in vivo can be determined.
  • a time-controlled mapping of known and newly identified disease-associated molecular markers in the whole organism can be carried out in vivo during the pathogenesis.
  • a function of the molecular markers in pathogenesis in vivo which has not yet been known can be determined.
  • new therapeutic drugs and / or treatment strategies can be derived from this knowledge by the fact that the determined surface states of cells (status of the cells with regard to the ac- tivierung / the rest state) regulated and these states are tracked with the inventive method.
  • the above methods can also be performed ex vivo on harvested organs and tissues.
  • mapping of the molecular markers in the organs and tissues from postembryonic ontogeny and during pathogenesis can of course be applied to various animal species and / or animal models, in particular in gerontology or for disease models.
  • the mapping can generally also be used for all animal species, for example agricultural livestock, domestic and domestic animals and wild animals, and for humans. Since mapping can be used in vivo, there is often no need for time-consuming and financially expensive conventional in-house processes, for example immunohistochemistry, which often have the additional disadvantage of having to identify and identify molecular markers at exactly one point in time to characterize.
  • the device according to the invention and the method according to the invention can also be used for the characterization of cell cultures by ro-mapping of molecular markers of cells of different origin that can be regulated up and down, for example under different cultivation conditions or for characterizing new cell lines or for re-characterizing existing ones Cell lines after different passages or immediately before their application, especially when implanting tumor cells in mice, rats and other animal models.
  • plant cells can also be characterized in addition to animal and human cells.
  • the bodies preferably bubbles
  • the bodies can be provided with binding molecules specific for the markers.
  • binding molecules specific for the markers.
  • a therapy progress check e.g. in chemotherapy
  • the bodies in particular bubbles, are usually contained in ultrasound contrast media and are preferably applied in the form of suspensions.
  • Bubbles with functionalized polyalkyl cyanoacrylates which are produced according to the process described in WO 01/68150 A1, are particularly preferred. The method described there is therefore expressly included in the disclosure content of the present application.
  • the polymers described in WO 01/68150 A1 are used to produce the bubbles preferred according to the invention. Therefore, these starting materials for producing the bubbles are expressly included in the disclosure content of the present application.
  • the method according to the invention can also be used for ultrasound contrast media which contain different types of body, preferably types of bubbles.
  • - type of shell of a body / bladder e.g. Material thickness, elasticity ,.
  • such bubble mixtures or their distribution patterns in the examination object can also be quantified with high spatial resolution using the method according to the invention, a) by sending different pulse shapes in succession at a position and / or receiving different characteristic signals; if the bubbles are not completely destroyed, a destruction pulse can be sent before moving to the next position; or, b) by the area to be examined being scanned several times with a correspondingly defined displacement, each scan being carried out with a different pulse shape required for the respective bubble type.
  • the bubbles differ simultaneously in the binding molecule on the surface and in their destructibility by sound, the following can be used to determine the in-vivo, ex-vivo and in-v / fro distribution of the binding sites in the examination region:
  • Bubble type A destructible with low sound pressure and with binding molecule
  • the saturation bubble concentration is increased by the factor in which the distance covered between two images or sound lobes is related to the layer thickness of the entire sound field. With a 10 ⁇ m shift and a layer thickness of 1 mm, the saturation bubble concentration increases by a factor of 100. At the same time, the layer thickness of the tissue from which the newly added bubbles originate is reduced by the same factor.
  • Histological resolution can be achieved in this way. Since this principle can also be used in vivo, one can speak of an “in wVo sono histology” or, in the case of a specific bladder accumulation, a sonographic “/ ⁇ -Vo immunohistology” with regard to the signals shown.
  • the measured MU can be evaluated for diagnosis as follows, for example:
  • Layer thickness and / or dose in a special measurement window (liver, tumor surface, vascular endothelium, etc.) based on the largest possible population.
  • the population can consist of individuals who do not differ in the distribution and the degree of enrichment of the bubbles in the measurement window, i.e. which is homogeneous or which is grouped in this regard. These dependencies generally have to be determined with various ultrasound device settings (transducer, detection mode, etc.).
  • Fig. 14 Dependence of the MU on the SAE layer thickness
  • Rat 1 (Example 2): 1 * 10 8 bubbles / ml; Ex vivo measurement, 30 min.
  • Rat 1 (Example 2): 1 * 10 8 bubbles / ml; Ex vivo measurement, 30 min.
  • Fig. 16 Dependence of the SAE CO r r on the SAE layer thickness rat 1 (example 2): 1 * 10 8 biases / ml; Ex vivo measurement, 30 min.
  • Fig. 17 Dependence of the MU on the SAE layer thickness
  • Rat 2 (Example 2): 1 * 10 7 bubbles / ml; Ex vivo measurement, 30 min.
  • Fig. 18 Dependence of the MU corr on the SAE layer thickness
  • Rat 2 (Example 2): 1 * 10 7 bubbles / ml; Ex vivo measurement, 30 min.
  • Rat 2 (Example 2): 1 * 10 7 bubbles / ml; Ex vivo measurement, 30 min.
  • Example 21 Color signals of the specific bubbles enriched in the tumor (four mice, example 3) in comparison to the isotype control (four mice, example 3).
  • Example 1 Color signals of the specific bubbles enriched in the tumor (four mice, example 3) in comparison to the isotype control (four mice, example 3).
  • Example 1 Color signals of the specific bubbles enriched in the tumor (four mice, example 3) in comparison to the isotype control (four mice, example 3).
  • the bubbles are made according to Example 4 (a).
  • the bubble concentration was adjusted to 6 * 10 8 bubbles / ml with 0.02% (m / m) Triton X 100 solution. Thereof were carefully stirred without introduction of air bubbles into the solution Agarlö- 33 ul (* 10 7 bubbles 2). The agar solution was then placed in a glass bowl
  • a transducer (L 10-5, ATL UM9, color Doppler mode, MI: 1, 1, persistence: 0, priority: maximum, size of the color box: maximum, depth of penetration: 3 cm, focus: 2 cm) was with a Tripod mounted vertically above the phantom so that it dipped into the water.
  • the pelvis was made together with the phantom 1. moved by hand in the transverse direction under the transducer at a speed of approx. 2 cm / s, which corresponded to a displacement distance of approx. 1 mm / frame, or
  • the video images were digitized at a sample rate corresponding to the frame rate given by the ultrasound device (here 5.8 frames / second) (QuantiCon).
  • a trigger delay of 172 ms 1000 ms / 5.8 was previously entered in QuantiCon.
  • FIG. 8 shows the 3D view of the scanned agar phantom as it was generated from the 3D data set in Quanticon the number of pixels of the gray values as well as the number of pixels of the segmented color values of all images in the data set were exported and further processed with MS-Excel ® (Microsoft).
  • MS-Excel ® MS-Excel ®
  • FIG. 9 The graphic of the segmented color values (MU) created in MS Excel is shown in FIG. 9 as a function of the SAE layer thickness. Each individual color value was corrected in accordance with the correction formula defined under “corrected MU”. The resulting dependence on the SAE layer thickness is shown in FIG. 10 (MU corr .).
  • the conversion factor F sae was first determined.
  • the sum of the SAE signals was first determined by counting by hand in 20 sectional images.
  • the sum of the segmented color pixels was then divided by the sum of the SAE effects counted by hand in the corresponding images.
  • An average of 44 color pixels per SAE signal was determined.
  • Table 1 The data on which FIGS. 9 to 11 are based are listed in Table 1: Table 1:
  • the curve range between 30 and 70 ⁇ m SAE layer thickness ( ⁇ 37 - 81% color saturation gung) can be described with a linear correlation function. Correlation coefficients are 0.990 for the uncorrected MU and 0.996 for the corrected MU / SAE.
  • the method according to the invention enables bubble quantification even with bubble concentrations greater than 3,000 bubbles / ml.
  • the saturation bubble concentration increases in the ratio in which the actual layer thickness of the sound field relates to the displacement distance or to the SAE layer thickness.
  • a 10 ⁇ m shift from image to image simultaneously increases the spatial resolution and the saturation bubble concentration by a factor of 100 with a layer thickness of the sound field of 1 mm.
  • rat 1 Two two rats (Wistar SCHOE, 220g, female) were examined. The rats were injected iv with various doses of the bladder suspension prepared according to a) via the tail vein. For rat 1 a dilution to 1 * 10 8 bubbles / ml, for rat 2 to 1 * 10 7 bubbles / ml in 0.9% (m / m) NaCl solution containing 0.02% (m / m ) Triton X 100.
  • Rat 1 1 * 10 8 bubbles / kg (1 * 10 8 bubbles / ml, 220 ⁇ l iv)
  • Rat 2 1 * 10 7 bubbles / kg (1 * 10 7 bubbles / ml, 220 ⁇ l iv)
  • the rats were injected i.p. Injection of 1 ml of a 1: 1 volume mixture of Rompun 2% (20 mg xylazine / ml) and Ketavet 100 mg / ml (100 mg ketamine base / ml) killed:
  • the right lobe of the liver was removed and placed in a vessel filled with 0.9% (m / m) sodium chloride solution.
  • the vessels with the liver lobes were:
  • Rat 1 (1 * 10 8 bubbles / kg): 15, 20, 30 ⁇ m / frame
  • Rat 2 (1 * 10 7 bubbles / kg): 20, 40, 80, 120 ⁇ m / frame
  • the ultrasound examination was carried out analogously to Example 1 with the same device settings.
  • the liver was moved past the transducer at a distance of 2 cm.
  • the same ROI was used for all measurements.
  • the size of the ROI (approx. 2 mm * 6 mm, measured on the scale of the ultrasound device) was chosen so that the ROI was completely in the area of the liver tissue for all evaluated images.
  • the same magnification was selected in Quanticon ® for all measurements.
  • a square ROI of 2 mm edge length (determined using the length measurement scale shown in the ultrasound image) was measured in the ultrasound image. For this purpose, the image was enlarged accordingly using the zoom function available in Quanticon ® . The pixel sum in this ROI (MU) was 225. From this, a ratio of Embedding 56.25 pixels / mm 2 (225/4) could be calculated. The bladder concentration / ml liver was then calculated from the measured volume (area of the ROI x displacement distance / frame). The following formula was used to convert the raw data (FP, GP, S) into the bladder concentration C in the tissue:
  • FP Screen pixels that lie within the measurement window (ROI) in the area of the SAE effects shown.
  • GP Screen pixels that lie within the measurement window (ROI) outside of the SAE effects shown.
  • ⁇ sae Sum of the SAE signals shown in an ultrasound image
  • F f sum of all screen pixels in mm 2 (measured using the length scale shown in the ultrasound image)
  • Tab. 2 also contains the mean values of the percentage color saturation for the respective SAE layer thickness.
  • the number of bubbles is in a linear relationship to the SAE layer thickness.
  • the method according to the invention can also be used in removed organs in which bubbles accumulate non-specifically. There is a close correlation between the number of SAE signals shown in the sectional view and the corresponding SAE layer thickness.
  • the bladder concentration in the livers of the rats examined differs significantly depending on the dose applied.
  • Example 4 (f) The specific bubbles were prepared and characterized according to Example 4 (a) to (e).
  • the antibody coupling was carried out according to Example 4 (f).
  • the isotype control bubbles were prepared and characterized according to Example 4 (a) to (e).
  • the isotype coupling was carried out according to Example 4 (g).
  • mice were given an ultrasound contrast agent, containing specific bubbles (produced and characterized according to Example 4 (a) to (e) and antibody coupling according to Example 4 (f)), in the waking state iv in a dose of 1 * 10 9 bubbles / kg body weight injected (corresponded to 200 ⁇ l of the suspension according to Example 4 (g) each 20 g mouse).
  • the specific bubbles were gas-filled microcapsules, the shell of which consisted of functionalized polybutyl cyanoacrylate and to whose surface anti-CD105 antibodies were coupled.
  • mice As a control substance, four further mice were injected in the same way with an ultrasound contrast agent containing non-specific bubbles (produced and characterized according to Example 4 (a) to (e) and isotype coupling according to Example 4 (g)).
  • the non-specific bubbles were gas-filled microcapsules, the shell of which also consisted of functionalized polybutyl cyanoacrylate, but to the surface of which IgG 2a (isotype control) was coupled.
  • the tumors of all animals were examined 60 minutes after application of the respective ultrasound contrast agent with regard to the degree of enrichment using the method according to Example 1.
  • mice were treated with a 1: 1 volume mixture of a dilution (1 + 9) of Rompun ® 2% (20 mg xylazine / ml) and a dilution (1 + 4) of Ketavet ® 100 mg / ml (100 mg Ketamine base / ml) anesthetized in physiological NaCl solution. 200 ⁇ l each of 20 g of mouse were injected ip from this anesthetic mixture.
  • the mouse was attached to a carrier that was attached to the moving part of a servo motor (Limes 150, OWIS GmbH, motor controller DC 500) with adhesive tape.
  • the servo motor was adjusted vertically with a tripod so that the carrier reached into a water basin (cut, square plastic bottle) with the mouse.
  • the pool was washed with fresh water at a temperature of 37 ° C crowded.
  • a transducer (L 10-5, ATL UM9) was provided with coupling gel and placed horizontally on the side wall of the pool with a tripod. The mouse was moved so far into the pelvis under visual and sonographic control that the tumor had not yet reached the sound field.
  • the transducer was switched on again and the animal was scanned again with the same device settings and the same SAE layer thickness a second scan was needed to identify color signals that were not caused by SAE signals and served as a control value, such signals could have been caused by flowing blood or larger air bubbles.
  • the animal's respiratory rate was generally low enough to be able to perform a scan of approx. 3 cm with the SAE layer thickness used here within one pause without movement artifacts (Fig. 20b).
  • breathing artifacts could be avoided as follows:
  • FIG. 20b A sectional image was generated in the scan direction (FIG. 20b) and this was superimposed on the same scale with the corresponding graphic of the measured values (FIG. 20a).
  • the sum of the colored screen pixels within the tumor was evaluated with Quanticon® (3D-Echotech). Only the color signals in the tumor (MUFP: 20) and not the gray signals (MUGP: 21) were taken into account for the evaluation.
  • Lines 16 and 17 drawn in FIGS. 20a / b delimit the tumor area during the first scan. The area of the second scan is shown between lines 18-19.
  • Reference number 20 color signals (MUFP), reference number 21: gray signals (MUGP), reference number 22: sum of MUFP and MU G p, or the measured tissue volume.
  • MUFP color signals
  • MUGP gray signals
  • reference number 22 sum of MUFP and MU G p, or the measured tissue volume.
  • the method according to the invention is also suitable for the quantification of specifically enriched bubbles in the living organism.
  • a significant difference can be detected even if scanning is not carried out with maximum spatial resolution and SAE saturation occurs in some areas of some sectional images.
  • the number of signals for a specific bladder accumulation can be distinguished from residual signals of the blood vessel system and / or signals of an unspecific bladder accumulation.
  • the particle size distribution of the microcapsule suspension according to Example 4 (a) was determined using a particle counter from Particle Sizing Systems, type AccuSizer 770 (measuring medium: aqueous 0.02% (m / m) Triton X100 solution).
  • the volume-weighted particle size distribution ranged from 0.8 to 10 ⁇ m with a maximum at 1.8 ⁇ m, and the microcapsule concentration was 7.2 * 10 9 ( ⁇ 1 * 10 8 ) particles per ml.
  • Example 4 2,500 g of a microcapsule suspension according to Example 4 (a) were mixed with stirring with 501 g of sodium hydroxide solution with a concentration of 8 * 10 "2 mol / l. In the reaction mixture the pH was 12. The mixture was stirred at room temperature for 20 minutes. The pH was then adjusted to pH 3.5 with 1 N hydrochloric acid.
  • the particle size distribution of the microcapsule suspension according to Example 4 (c) was determined using a particle counter from Particle Sizing Systems, type AccuSizer 770 (measuring medium: aqueous 0.02% (m / m) Triton X100 solution).
  • the volume-weighted particle size distribution ranged from 0.8 to 10 ⁇ m with a maximum at 1.8 ⁇ m, and the microcapsule concentration was 6.0 * 10 9 ( ⁇ 1 * 10 8 ) particles per ml.
  • the mixture was then stirred for an hour with occasional pH control.
  • the mixture was stirred further at 4 ° C. overnight (approx. 17 h).
  • the suspension of gas-filled microbubbles according to Example 4 (a) to (d) to which streptavidin according to Example 4 (e) had been bound was treated with biotinylated anti-CD105 antibody (Rat Anti-Mouse CD105 / Endoglin, Biotin Conjugate, Southern Biotechnology Associates, Inc., Cat. No. 1860-08) in a ratio of 0.5 ⁇ g antibody to 1e6 microbubbles. After incubation, the suspension was diluted to 1e8 bubbles / ml with PBS buffer containing 0.02% (m / m) Triton x 100. The CD105 coupling and dilution was freshly made for each animal.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Biophysics (AREA)
  • Medical Informatics (AREA)
  • Molecular Biology (AREA)
  • Radiology & Medical Imaging (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Nanotechnology (AREA)
  • Epidemiology (AREA)
  • Surgery (AREA)
  • Pathology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Hematology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Acoustics & Sound (AREA)
  • Nonlinear Science (AREA)
  • Computer Networks & Wireless Communication (AREA)
  • General Physics & Mathematics (AREA)
  • Radar, Positioning & Navigation (AREA)
  • Remote Sensing (AREA)
  • Immunology (AREA)
  • Ultra Sonic Daignosis Equipment (AREA)

Abstract

L'invention vise à remédier au problème rencontré lors de la quantification de corps, notamment de bulles, dans les diagnostics ultrasoniques, à savoir que la concentration de corps, notamment de bulles, est souvent si grande que les données image obtenues sont saturées en ce qui concerne leur représentation, ce qui rend impossible leur quantification. A cet effet, les corps contenus dans les couches de coupe transversale d'un objet à examiner sont stimulées à l'aide d'ultrasons pour émettre des signaux caractéristiques. Ensuite, ces signaux sont enregistrés, des jeux de données sont formés à partir de ces signaux, les jeux de données sont transformés en une représentation du système de corps dans l'objet à examiner et le nombre nombre de corps est déterminé. Au moins deux jeux de signaux sont enregistrés à partir des couches de coupe transversale chevauchantes dans l'objet à examiner.
PCT/EP2003/003227 2002-03-28 2003-03-27 Dispositif et procede de quantification de corps par des ultrasons Ceased WO2003082117A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP03745194A EP1487345A1 (fr) 2002-03-28 2003-03-27 Dispositif et procede de quantification de corps par des ultrasons
AU2003232191A AU2003232191A1 (en) 2002-03-28 2003-03-27 Device and method for quantifying bodies by means of ultrasound
JP2003579664A JP2005527270A (ja) 2002-03-28 2003-03-27 超音波により物体を定量するための装置及び方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10215335.3 2002-03-28
DE10215335A DE10215335A1 (de) 2002-03-28 2002-03-28 Vorrichtung und Verfahren zur Quantifizierung von Blasen

Publications (1)

Publication Number Publication Date
WO2003082117A1 true WO2003082117A1 (fr) 2003-10-09

Family

ID=28051185

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2003/003227 Ceased WO2003082117A1 (fr) 2002-03-28 2003-03-27 Dispositif et procede de quantification de corps par des ultrasons

Country Status (5)

Country Link
EP (1) EP1487345A1 (fr)
JP (1) JP2005527270A (fr)
AU (1) AU2003232191A1 (fr)
DE (1) DE10215335A1 (fr)
WO (1) WO2003082117A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8366625B2 (en) 2006-03-10 2013-02-05 Mcgill University Ultrasound molecular sensors and uses thereof
US10585185B2 (en) 2017-02-03 2020-03-10 Rohde & Schwarz Gmbh & Co. Kg Security scanning system with walk-through-gate

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5860931A (en) * 1997-10-10 1999-01-19 Acuson Corporation Ultrasound method and system for measuring perfusion
US5947904A (en) * 1997-08-21 1999-09-07 Acuson Corporation Ultrasonic method and system for imaging blood flow including disruption or activation of a contrast agent
US6302846B1 (en) * 1999-09-20 2001-10-16 Acuson Corporation Ultrasound method for assessing ejection fraction using ultrasound contrast agents

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5425366A (en) * 1988-02-05 1995-06-20 Schering Aktiengesellschaft Ultrasonic contrast agents for color Doppler imaging
DE19813174A1 (de) * 1998-03-25 1999-05-27 Schering Ag Mikropartikel aus Polymeren und mindestens einer gerüstbildenden Komponente und ihre Herstellung und Verwendung in der Ultraschalldiagnostik und zur ultraschallinduzierten Wirkstofffreisetzung
US6186951B1 (en) * 1998-05-26 2001-02-13 Riverside Research Institute Ultrasonic systems and methods for fluid perfusion and flow rate measurement
US5971928A (en) * 1998-11-02 1999-10-26 Acuson Corporation Diagnostic medical ultrasonic system and method for image subtraction
US6340348B1 (en) * 1999-07-02 2002-01-22 Acuson Corporation Contrast agent imaging with destruction pulses in diagnostic medical ultrasound

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5947904A (en) * 1997-08-21 1999-09-07 Acuson Corporation Ultrasonic method and system for imaging blood flow including disruption or activation of a contrast agent
US5860931A (en) * 1997-10-10 1999-01-19 Acuson Corporation Ultrasound method and system for measuring perfusion
US6302846B1 (en) * 1999-09-20 2001-10-16 Acuson Corporation Ultrasound method for assessing ejection fraction using ultrasound contrast agents

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8366625B2 (en) 2006-03-10 2013-02-05 Mcgill University Ultrasound molecular sensors and uses thereof
US10585185B2 (en) 2017-02-03 2020-03-10 Rohde & Schwarz Gmbh & Co. Kg Security scanning system with walk-through-gate

Also Published As

Publication number Publication date
AU2003232191A1 (en) 2003-10-13
EP1487345A1 (fr) 2004-12-22
JP2005527270A (ja) 2005-09-15
DE10215335A1 (de) 2003-10-16

Similar Documents

Publication Publication Date Title
US6872180B2 (en) Device and process for quantifying bodies by means of ultrasound
Goertz et al. Subharmonic contrast intravascular ultrasound for vasa vasorum imaging
DE69319438T2 (de) Verbesserungen an kontrastmitteln
DE69729579T2 (de) Polymer-lipide mikroverkapselte gase und ihre anwendung bei bilderzeugenden mitteln
DE69635516T2 (de) Verwendung von Emulsionen enthaltend in lipide eingeschlossene flussige Fluorocarbone für gezielte diagnostische Verfahren in vivo
DE69732568T2 (de) Druckbeständige protein-mikrosphäre als ultraschallkontrastmittel
KR20150099823A (ko) 시간 경과에 따른 통계 분석에 기초한 영상 진단 애플리케이션에서의 분할
Tiemann et al. Stimulated acoustic emission: pseudo-Doppler shifts seen during the destruction of nonmoving microbubbles
CN107080744A (zh) 合成空心球的方法
JP2012523904A (ja) 高周波数における超音波造影剤の非線形撮像のための方法
WO2013034709A1 (fr) Dosimétrie de rayonnement in situ non invasive
Yoon et al. Angled-focused 45 MHz PMN-PT single element transducer for intravascular ultrasound imaging
Niżański et al. B-mode and Doppler ultrasonographic findings of prostate gland and testes in dogs receiving deslorelin acetate or osaterone acetate
Hughes et al. Characterization of digital waveforms using thermodynamic analogs: detection of contrast-targeted tissue in vivo
Reinhardt et al. Ultrasound derived imaging and quantification of cell adhesion molecules in experimental autoimmune encephalomyelitis (EAE) by Sensitive Particle Acoustic Quantification (SPAQ)
Rix et al. Evaluation of high frequency ultrasound methods and contrast agents for characterising tumor response to anti-angiogenic treatment
Grishenkov et al. In vitro contrast-enhanced ultrasound measurements of capillary microcirculation: Comparison between polymer-and phospholipid-shelled microbubbles
US8192362B2 (en) Methods of monitoring cellular death using low frequency ultrasound
Casciaro et al. Effectiveness of functionalized nanosystems for multimodal molecular sensing and imaging in medicine
DE4219724A1 (de) Verwendung von Mikrokapseln als Kontrastmittel für die Farbdoppler-Sonographie
EP1487345A1 (fr) Dispositif et procede de quantification de corps par des ultrasons
Linker et al. In vivo molecular imaging of adhesion molecules in experimental autoimmune encephalomyelitis (EAE)
DE60310429T2 (de) Verfahren zur identifizierung von wächter-lymphknoten
JP2022513542A (ja) 高感度分子解析のための組成及び方法
Miller et al. Diagnostic ultrasound-induced membrane damage in phagocytic cells loaded with contrast agent and its relation to Doppler-mode images

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NI NO NZ OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2003745194

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2003579664

Country of ref document: JP

WWP Wipo information: published in national office

Ref document number: 2003745194

Country of ref document: EP