System and method for measuring at least one property of a particle using a mechanically coupled mechanical resonator and gravimetric sensor.

Abstract

The invention relates to a system for measuring at least one property of a particle (P), comprising: A mechanical resonator (1) having at least a first fixed part (10) and a first suspended part (11) capable of vibrating relative to the first fixed part, and a fluidic circuit integrated in its suspended part, through which a fluid containing said particle (P) is circulated; Excitation means (4) configured to vibrate the first suspended part (11) at an excitation frequency (F_smr); A first gravimetric sensor (2) comprising at least a second fixed part (20) and a second suspended part (21) capable of vibrating relative to the second fixed part (20); The first suspended part (11) of the mechanical resonator (1) and the second suspended part (21) of the first gravimetric sensor (2) being mechanically coupled to each other via a mechanical linkage element (3). Figure to be published with the abbreviation: Figure 4A

Description

Title of the invention: System and method for measuring at least one property of a particle using a mechanical resonator and a mechanically coupled gravimetric sensor. Technical field of the invention

[0001] The present invention relates to a system and method for measuring at least one property of a particle using a mechanical resonator with an integrated fluidic channel and a gravimetric sensor mechanically coupled. State of the art

[0002] To detect the presence of biomarkers of interest such as proteins, exosomes, circulating RNA, circulating DNA, viruses, or larger target species such as bacteria and cells, it is known to use biosensors.

[0003] In biosensors, there is in particular the category of gravimetric sensors. These are based on the use of a mechanical oscillator or resonator, equipped with a suspended part set into vibration at its resonance frequency. Any target that is grafted onto the surface of the suspended part of the resonator, causes the mass of the latter to increase, which decreases its resonance frequency, by an offset proportional to the mass of the captured target.

[0004] By continuously measuring the resonance frequency fluctuations, it is then possible to trace the mass adsorbed on the resonator in real time and, for example, to follow adsorption kinetics with targets.

[0005] A solution proposed in the state of the art consists of integrating and delimiting a fluidic channel within the suspended part of the resonator, while the latter oscillates in a cavity free of fluid. The advantage then lies in a very little altered quality factor, even in the presence of fluid circulating in the oscillator, and thus an optimized detection limit. This type of resonator is commonly called SMR ("Suspended MicroChannel Resonator") or SNR ("Suspended Na-nochannel Resonator"), depending on the dimensions of its fluidic channel. It can be noted that a resonator equipped with a fluidic channel whose width or thickness is less than Ipm will be referenced as an SNR, otherwise it will be an SMR.

[0006] By a pressure gradient imposed between a fluid inlet and a fluid outlet of the circuit, it is possible to control the flow of the fluid (its flow rate and direction of flow), and therefore the passage of particles in suspension in the SMR (or SNR). This type of sensor has been used for various applications, including the individual weighing of biological particles such as cells, bacteria, nanoparticles or even the detection of specific proteins by the prior functionalization of the internal walls of the SMR. This operating principle is now well known. Patent application US2021 / 046477A1 and patents US8899102B1, US8631685B2 and US8312763B2 describe this type of sensor.

[0007] Although the solution using an SNR type resonator has an improved detection limit compared to that using an SMR type resonator, it has certain limitations: - The dimensions of the suspended part of the SNR lead to limitations on the size of its fluidic channel and therefore on the size of the particles that can be injected and analyzed; - These dimensional constraints may require prior preparation of the sample to be analyzed, so that the particles can circulate freely in the fluidic channel of the SNR resonator without the risk of it becoming blocked due to the presence of particles larger than its fluidic section; in other words, the resonator is not necessarily suitable for the analysis of complex samples (such as blood for example, because it contains both cells and proteins, covering a size dynamic ranging from nanometers to several micrometers for cells typically 10 to 15qm in diameter);

[0008] There is therefore a need to have a solution that makes it possible both to release the dimensional constraints of the fluidic circuit of an SNR type resonator and to achieve a detection limit at least equivalent to that of an SNR type resonator. Statement of the invention

[0009] This aim is achieved by a system for measuring at least one property of a particle, comprising: - A mechanical resonator comprising at least a first fixed part and a first suspended part capable of vibrating relative to the first fixed part, and a fluid circuit integrated in its suspended part, in which a fluid containing said particle is caused to circulate, - Excitation means configured to vibrate the first suspended part at an excitation frequency, - A first gravimetric sensor comprising at least a second fixed part and a second suspended part capable of vibrating relative to the second fixed part at a vibration frequency, - The first suspended part of the mechanical resonator and the second suspended part of the first gravimetric sensor being mechanically coupled to each other via a mechanical connecting element, - Means for measuring variations in the vibration frequency of the second suspended part of the first gravimetric sensor.

[0010] Advantageously, the excitation frequency corresponds to the resonant frequency of the resonator or that of a higher natural mode of this resonant frequency.

[0011] According to an advantageous embodiment, the mechanical connecting element comprises a first end secured to the first suspended part at a first point corresponding to a vibration antinode of the first suspended part, when the latter is excited at its resonance frequency or at that of a higher natural mode.

[0012] According to an advantageous embodiment, the mechanical connecting element comprises a second end secured to the second suspended part on a second point corresponding to a vibration anti-node of the second suspended part, when the latter is excited at the vibration frequency.

[0013] According to a particular embodiment, the first suspended part is chosen from a beam embedded on a single fixed part, a beam embedded between two fixed parts, and a plate held between several fixed parts.

[0014] According to a particular embodiment, the second suspended part is chosen from a beam embedded on a single fixed part, a beam embedded between two fixed parts, and a plate held between several fixed parts.

[0015] According to a particular feature, the excitation means are configured to vibrate the second suspended part at a vibration frequency close to the resonance frequency of the first suspended part or to that of a higher natural mode, at more or less 30% of said resonance frequency or that of the higher natural mode.

[0016] According to one particular feature, the excitation means comprise at least one piezoceramic element.

[0017] According to another particularity, the measuring means comprise one or more piezoresistive gauges.

[0018] According to another feature, the fluidic circuit integrated into the first suspended part of the mechanical resonator comprises a main fluidic channel and a hydrodynamic trap positioned in its channel and intended to trap said particle.

[0019] According to another particular embodiment, the system comprises: - A second gravimetric sensor, comprising at least a third fixed part and a third suspended part capable of vibrating relative to the third fixed part at a second vibration frequency, - The first suspended part of the mechanical resonator and the third suspended part of the second gravimetric sensor being mechanically coupled to each other via a second mechanical connecting element, - Second means for measuring variations in the second vibration frequency of the third suspended part of the second gravimetric sensor.

[0020] According to a particular feature, the third suspended part is chosen from a beam embedded on a single fixed part, a beam embedded between two fixed parts, and a plate held between several fixed parts.

[0021] The invention also relates to a method for measuring at least one property of a particle, implemented using a measuring system as defined above, the method comprising the following steps: - Injection of a fluid containing said particle into the fluid circuit of the mechanical resonator, - Excitation of the first suspended part of the mechanical resonator at its resonance frequency, - Excitation of the second suspended part of the first gravimetric sensor at the vibration frequency, - Measurement of variations in the vibration frequency of the second suspended part of the first gravimetric sensor.

[0022] According to one feature, the method comprises a step of calibrating the system by injecting into the fluid circuit a fluid containing one or more calibrated particles.

[0023] The principle of the invention is thus based overall on the mechanical coupling of an oscillating structure carrying a fluidic channel (i.e. an SMR), with a solid oscillator (without a fluidic channel) which is much smaller (and which will be called NEMS for Nano Electro-Mechanical System or otherwise MEMS for Micro Electro-mechanical System depending on the dimensions), and which is used to measure frequency fluctuations following the passage of particles or targets circulating in the fluidic channel.

[0024] In the context of the invention, it is thus possible to exploit the small dimensions of the gravimetric sensor (i.e. the NEMS or MEMS), by minimizing its effective mass (proportional to its volume, to the density of the material which constitutes it and which is also a function of the resonance mode). It is possible to obtain an optimized detection limit compared to that of the mechanical resonator including the fluidic channel (i.e. the SMR).

[0025] At the same time, the micrometric dimensions of the fluidic channel integrated into an SMR type resonator make it possible to circulate targets or particles of micrometric size (typically 10 to 15 μm in diameter for cells, or even more), as well as more complex fluids than in the case of standard SNR type resonators, the latter being constrained in dimensions by the sub-micrometric width (i.e. less than 1 μm) of their fluidic channel.

[0026] Generally speaking, the mechanical coupling of the SMR resonator with the NEMS sensor will be carried out in such a way that the oscillation drive of the suspended part of the NEMS sensor by the suspended part of the SMR resonator is maximized, and to minimize the damping of the vibrations of one and the other, thus optimizing the quality factor. The challenge is that the mechanical coupling damps the vibrations of the NEMS sensor as little as possible, and vice versa those of the SMR resonator, and therefore that the most efficient transduction possible is obtained between the two coupled oscillators. Brief description of the figures

[0027] Other characteristics and advantages will appear in the detailed description which follows, given with reference to the appended drawings in which: Figures 1A and 1B represent, respectively seen in perspective and seen from above, a mechanical resonator of the SMR type; [Fig.2] shows examples of deformation curves of a mechanical resonator of the SMR type according to different vibration modes; Figures 3A and 3B represent, respectively seen in perspective and seen from above, a NEMS type sensor; Figures 4A and 4B show a particular embodiment of the measuring system of the invention and illustrate its operating principle; Figures 5 to 11 show several variant embodiments of the measuring system of the invention; [Fig. 12] shows the steps of an exemplary manufacturing method of the measuring system of the invention;

[0028] Detailed description of at least one embodiment

[0029] The invention relates to a system used for measuring at least one property (for example mass, volume, density) of a particle. By particle is meant for example a biological particle such as cell, exosome, virus, bacteria... By particle is also meant a particle of inorganic type such as for example a particle of Gold, Polystyrene...

[0030] The system of the invention is based on the use of at least two entities mechanically coupled together. Mechanical Resonator SMR or SNR

[0031] [Fig.lA] [Fig.lB] [Fig.2]

[0032] A first entity of the system consists of a mechanical resonator 1, better known under the name SMR ("Suspended MicroChannel Resonator" - hereinafter referred to as also SMR resonator) or SNR ("Suspended Nanochannel Resonator"), its micro or nano character depending in particular on the dimensions of its integrated fluidic channel. A resonator equipped with a fluidic channel whose width or thickness is less than Ipm will be referenced as an SNR, otherwise it will be an SMR.

[0033] Indeed, such a mechanical resonator 1 (SMR or SNR) comprises at least one fixed part 10, and a suspended part 11 connected to the fixed part 10 and capable of vibrating when it is excited. The suspended part 11 is conventionally in the form of a beam extending along a so-called longitudinal axis.

[0034] It also comprises a fluidic circuit integrated into its suspended part, this fluidic circuit comprising at least one fluidic channel 12 dug inside its suspended part 11, covered with sealed walls on the four faces (above, below, laterally) and intended to be used to inject a fluid into it in which one or more particles P to be characterized are placed.

[0035] Advantageously, the mechanical resonator 1 is of the SMR type (rather than an SNR), so as to be able to circulate larger sized particles P therein, and to analyze complex samples (see below).

[0036] The system comprises excitation means 4 configured to vibrate the suspended part 11 of the mechanical resonator 1 at an excitation frequency F_smr. This excitation frequency is advantageously the resonant frequency of the resonator or a frequency equivalent to a higher mode of this resonant frequency.

[0037] In this type of resonator, the vibration is generally carried out out of plane (as indicated by the curved arrow in [Fig.lA]), but alternatively it can be generated in the plane, or even in torsion. The in-plane or out-of-plane vibration is carried out in a horizontal or vertical plane respectively when the longitudinal axis of the beam is positioned horizontally.

[0038] The suspended part 11 may be simply fitted, connected to a single fixed part and thus provided with a free cantilevered end. In this example, we will speak of a "cantilever".

[0039] The suspended part 11 can also be double-mounted, thus joining two fixed parts.

[0040] The suspended part 11 can also be produced in the form of a plate supported by several anchor points.

[0041] Whatever its architecture, this suspended part is likely to be set into vibration to oscillate at the excitation frequency F_smr, this frequency advantageously being the resonance frequency at its fundamental natural mode or one of its higher natural modes, or even a combination of these modes.

[0042] As a reminder, such a mechanical resonator 1 can be excited according to several vibration modes, designated for example M_1, M_2, M_3 in [Fig.2]. In this example, it This is an out-of-plane bending vibration.

[0043] For each vibration mode, the resonator has vibration nodes and vibration antinodes.

[0044] As a reminder, a vibration anti-node corresponds to an area of ​​the suspended part where the vibration amplitude is maximum for the requested vibration mode.

[0045] [Fig.2] indicates the presence of vibration antinodes A_l, A_2, A_3 for each of the three vibration modes.

[0046] By a pressure gradient imposed between the ports located upstream and downstream of the fluidic channel 12 integrated into the suspended part 11, it is possible to control the flow of the fluid (its flow rate and direction of flow) in the fluidic channel 12, and therefore the passage of particles P in suspension in the fluidic channel of the mechanical resonator 1. When a particle P circulates in the fluidic channel of the resonator, it transiently alters the mass (of Am) of the suspended part 11 of the resonator 1, causing a shift in its (resonant) frequency which is proportional to the floating mass of the particle.

[0047] The shift of the frequency AF_smr ([Fig. 1 A]) of the resonator depends in fact on the added mass Am, the total mass m of the resonator as well as a correction coefficient a which depends specifically on the position of the added particle P, according to the following relation:

[0048] &F smr _ km F_smr m

[0049] Thus, when a particle of given floating mass Am is injected into the fluidic channel 12 of the resonator, a maximum vibration amplitude is obtained when this particle P is located at the level of the vibration antinode (for the vibration mode M_1 for example) and therefore a maximum frequency shift. Note that, depending on the difference in density between the particle and the carrier fluid, the floating mass Am will be positive (in the case of a denser particle) or negative (in the case of a less dense particle). Gravimetric sensor

[0050] - [Fig.3A] [Fig.3B]

[0051] The second entity of the system consists of a gravimetric sensor 2 (hereinafter also referred to as a NEMS sensor). For example, this gravimetric sensor 2 is advantageously produced in the form of a NEMS (Nano Electro-Mechanical System).

[0052] This type of gravimetric sensor is also based on a mechanical oscillator, set in vibration at a vibration frequency F_nems, and without a fluid channel.

[0053] It thus comprises a fixed part 20 and a suspended part 21 connected to its fixed part and capable of vibrating relative to the fixed part 20 when it is excited.

[0054] The suspended part 21 is also conventionally in the form of a beam extending along a so-called longitudinal axis.

[0055] Unlike the mechanical resonator 1, responsible for circulating the particles P in its fluidic channel 12, the gravimetric sensor 2 does not include a fluidic circuit. It is only used as a means of measuring frequency fluctuations (see below).

[0056] The excitation means 4 of the system are configured to vibrate the suspended part 21 of the gravimetric sensor 2 at the vibration frequency F_nems. It will be seen that the vibration of the NEMS sensor can be achieved via the mechanical resonator 1 and the mechanical connection between the two elements, without external excitation means.

[0057] In this type of sensor, the vibration is generally carried out out of plane (as indicated by the solid arrow in [Fig.3A]) or alternatively in plane (as indicated by the dotted arrow in [Fig.3A]). A vibration variant can be carried out in a torsion mode. The in-plane or out-of-plane vibration is carried out in a horizontal or vertical plane respectively when the longitudinal axis of the beam is positioned horizontally.

[0058] The vibration frequency F_nems is for example equal to a value which can range from -30% to +30% of the value of the excitation frequency F_smr of the mechanical resonator, advantageously its resonance frequency or that of its higher natural mode.

[0059] This vibration frequency F_nems is advantageously chosen to be close to the resonance frequency F_smr of the mechanical resonator 1 or that of its higher natural mode in order to harmonize the vibration of the two suspended entities. Mechanical coupling

[0060] - [Fig.2] [Fig.4A] [Fig.4B]

[0061] The principle of the invention is based on a mechanical coupling between a mechanical resonator 1 as described above with a gravimetric sensor 2 as described above.

[0062] The mechanical coupling is achieved by integrating a mechanical connection element 3 between the suspended part 11 of the mechanical resonator and the suspended part 21 of the gravimetric sensor. This mechanical connection element 3 is for example a simple beam forming the junction between the two suspended parts. It may in particular be one or more nano-beams obtained according to the manufacturing method implemented.

[0063] The mechanical resonator 1 is thus used to convey the particles P in circulation in its fluidic channel 12. It is set into vibration at its excitation frequency F_smr (advantageously its resonance frequency or a higher eigenmode thereof). The gravimetric sensor 2 is also excited at the vibration frequency F_nems, advantageously close to the excitation frequency of the mechanical resonator and is used to measure the frequency fluctuations at the level of the resonator 1 when the latter receives a particle in its fluidic channel 12.

[0064] The mechanical coupling of the mechanical resonator 1 (advantageously of the SMR type) with the gravimetric sensor 2 (advantageously of the NEMS type) and the reading of the frequency fluctuations at the gravimetric sensor 2 make it possible to: - Cover a wider range of particles P than when using a mechanical resonator alone (for example an SNR), in particular in terms of size of objects to be weighed; the fluidic channel 12 integrated into the SMR type resonator is enlarged (for example 20x20qm2 for an SMR resonator against 0.7x0.7qm2 for an SNR type resonator, i.e. a gain of approximately 816 in terms of surface ratio). The corollary of this increase in the fluidic section is the reduction in the fluidic resistance and therefore the increase in the flow rate at identical pressure gradient. - Reduce the risks of clogging inherent in the nano-fluidic channels of SNR type resonators, - Offer a mass detection limit comparable to that of SNR type resonators (currently attogram detection limit) thanks to the reading of frequency fluctuations at the level of the NEMS gravimetric sensor mechanically coupled to the SMR type resonator.

[0065] The beam (or suspended part) 12 of the mechanical resonator 1 is capable of being excited according to several vibration modes (for example M_l, M_2, M_3), each of these modes resulting in a deformation of the beam 12 locally presenting vibration nodes and antinodes (also called antinodes) at different locations predetermined by the laws of structural mechanics and dynamics. As a reminder, a vibration antinode A_l, A_2, A_3 corresponds to a zone where the vibration amplitude is maximum.

[0066] The mechanical coupling of the NEMS 2 sensor with the SMR resonator is advantageously designed to maximize the transmission of vibration waves between the two oscillators, and to limit as much as possible the damping of the oscillations of one and the other, generated by this coupling.

[0067] It is important that any fluctuation AF_smr of the excitation frequency of the SMR resonator 1 results in a variation AF_nems of frequency at the NEMS sensor 2 ([Fig.4B]). An example consists of arranging the mechanical connection at a vibration anti-node of the SMR resonator 1 on the vibration mode for which the latter is actuated in order to maximize the transduction between it and the NEMS sensor. Means of excitement

[0068] - [Fig.4A]

[0069] It should be noted that several transduction / excitation modes are possible for: - Actuating the SMR resonator and the NEMS gravimetric sensor at the chosen frequency (for example resonance frequency for the SMR and vibration frequency for the NEMS), and - Allow the measurement of frequency and its fluctuations at the NEMS gravimetric sensor.

[0070] The vibration of the NEMS sensor can be obtained solely and directly via the mechanical connection which exists between the suspended part 11 of the mechanical resonator 1 and the suspended part 21 of the NEMS sensor. It can also be obtained by controlling the excitation means 4, configured to excite both (see below the advantageous combination) the suspended part 11 of the mechanical resonator 1 at the excitation frequency (for example fundamental resonance frequency or a higher natural mode thereof) and the suspended part 21 of the NEMS sensor at the desired vibration frequency.

[0071] Preferably, an advantageous combination is based on: - Actuation / excitation by piezoelectric effect (for example by bringing a piezoceramic into contact with the rear face of the component) to simultaneously actuate the SMR resonator and the NEMS sensor. The piezoceramic is glued to a printed circuit, itself designed to apply an electrical potential to it which will deform the piezoelectric material. The latter is made to vibrate by applying, for example, an electrical signal at the resonance frequency of the SMR or NEMS sensor, or a signal comprising the different frequency signals corresponding to the modes of the oscillators present on the chip. The advantage of this configuration lies in the fact that the applied signal can collectively actuate the different oscillators present on the component (NEMS and SMR) and the different resonance modes desired for each of them. This is therefore an approach that is perfectly suited to an oscillator network. - A reading by piezoresistive effect by implanting for example the NEMS layer with dopants and a suitable concentration, either locally to delimit a gauge at the embedding of the NEMS sensor or on the full surface by exploiting the structuring of suspended nano-strain gauges connecting for example the NEMS sensor, either to the SMR resonator or to fixed pads.

[0072] Actuation by capacitive coupling is also a possible solution. In this case, it is necessary to integrate an electrode as close as possible to the NEMS sensor 2 and the SMR 1 resonator if it also needs this actuation source. This transduction mode is well suited for lateral vibration modes but requires that the suspended oscillating part and the actuation electrode which is located opposite are sufficiently conductive so that a transduction capacity can be available.

[0073] In [Fig.4A], the localization zone of a piezoresistor PZ_1 (via doping - for example with Boron) is shown at the level of the suspended part 11 of the SMR resonator 1 and of the suspended part 21 of the NEMS sensor, this zone ending with the presence of a piezoresistive gauge PZ_2 in extension of the connecting element 3 between the SMR resonator and the NEMS sensor. This solution makes it possible to determine the frequency variations at the level of the NEMS sensor 2, because the current variations in the piezoresistor will be maximum at the resonance frequency of the NEMS sensor. System implementation variants

[0074] - Figures 4A to 11

[0075] Based on these principles, the mechanical connection between the beam of the SMR resonator and that of the NEMS sensor can be made according to different configurations. It should also be noted that it is possible to mechanically couple a single SMR resonator to several NEMS sensors (for example two NEMS sensors). The measuring means (for example via piezoresistivity - PZ_1 + PZ_2) are not shown in Figures 5 to 11 but it should be considered that they are of course present.

[0076] It should be noted that in all configurations, the two suspended parts 11, 21 extend, along their longitudinal axis, in parallel to each other and the connecting element 3 extends, along its longitudinal axis, perpendicular to the two suspended parts and thus creates a junction between the two suspended parts.

[0077] Figures 4A to 11 illustrate several possible configurations. These are to be considered in a non-limiting manner. In each configuration, on the NEMS sensor 2 side, the mechanical connection element 3 is fixed at a vibration anti-node for one of its vibration modes. And on the SMR resonator 1 side, the mechanical connection element 3 is fixed in a zone of its suspended part which corresponds to an anti-node for one or more vibration modes. It should be noted that the beam forming the mechanical connection element can be formed of several nano-beams. - [Fig.4A] - An SMR resonator + a NEMS sensor: A single-embedded beam 11 of the SMR resonator 1 is mechanically coupled to the beam 21 of a NEMS sensor by a lateral mechanical connection. In this example, the mechanical connection element 3, formed by a coupling beam, is located at the first antinode of the beam 11 of the SMR resonator since its embedding, for its second mode of vibration in bending outside plan. [Fig.5] - An SMR resonator + a NEMS sensor: A single-embedded beam 11 of the SMR resonator 1 is mechanically coupled to the beam 21 of a NEMS sensor by a lateral mechanical connection. In this example, the mechanical connection element 3, formed by a coupling beam, is located at the free end of the beam 11, i.e. at an antinode for its first, second and third out-of-plane bending vibration mode. [Fig.6]: One SMR resonator + one or two NEMS sensors: A single-embedded beam of the SMR resonator is mechanically coupled to the beam of one NEMS sensor or each of the two NEMS sensors, by a lateral mechanical connection located at its free end, either at an antinode for its first, second and third out-of-plane bending vibration mode. The same references are kept for the two NEMS sensors. [Fig.7]: One SMR resonator + one or two NEMS sensors: A single-embedded beam of the SMR resonator is mechanically coupled to the beam 21 of one NEMS sensor or each of the two NEMS sensors, by a lateral mechanical connection, located at the first antinode of the SMR resonator beam from its embedding for its second out-of-plane bending vibration mode. [Fig.8]: One SMR resonator + two NEMS sensors: A single-embedded beam of the SMR resonator is mechanically coupled to the beam of each of the two NEMS sensors, by a first lateral mechanical connection located at its free end, and by a second mechanical connection at its first anti-node of the SMR beam from its embedding for its second out-of-plane bending vibration mode. [Fig.9]: SMR resonator + a NEMS sensor: The beam of the SMR resonator is double-embedded (between two fixed parts 10) and mechanically coupled to a NEMS sensor by a lateral mechanical connection located on an anti-node for its first vibration mode in out-of-plane bending (or in the plane as well). [Fig. 10] - An SMR resonator + a NEMS sensor: A single-embedded beam 11 of the SMR resonator 1 is mechanically coupled to the beam 21 of a NEMS sensor by a lateral mechanical connection. In this example, the mechanical connection element 3, formed by a single coupling beam, is located at the first antinode of the SMR resonator beam from its embedding for its second out-of-plane bending vibration mode. In addition, the NEMS sensor is made in the form of a bi- embedded (between two fixed parts 20). The connecting element 3 is connected to this beam on an anti-node of the NEMS sensor for its first vibration mode in out-of-plane bending (or in the plane as well). - [Fig.l 1] - An SMR resonator + a NEMS sensor: The fluidic circuit of the SMR resonator has one or more trapping zones, each dedicated to receiving a particle. By this means it is possible to trap one or more particles (for example cells) in the fluidic circuit and to follow in particular, with the precision of the NEMS 2 sensor coupled to it, different parameters such as the mass growth rate of the particle during its exposure to a drug (to evaluate its effectiveness), the growth of the particle in a medium or the secretion of viral particles or exosomes. Instead of a cell, one (or more) functionalized beads (covered with antibodies or aptamers) can be trapped and the grafting kinetics followed with targets such as proteins for example.The hydrodynamic trap is produced in the form of a branch branch relative to the fluidic channel 12 of the SMR resonator 1 and comprises a housing 120 sized to accommodate the particle P and a restriction 121 extending said housing.

[0078] A combination of the characteristics of the different embodiments described above is also conceivable. Special achievement

[0079] In a non-limiting manner, the SMR resonator 1 and the NEMS sensor 2 are chosen to have a free-standing beam ("cantilever" - a suspended part connected to a single fixed part). The length and thickness of the NEMS sensor 2 are chosen so that its fundamental frequency according to the first vertical bending mode coincides with the resonance frequency for the second vertical bending mode of the SMR resonator.

[0080] The table below gives an example of sizing: Length and width (pm) Width and depth (pm) integrated channel Width of interior wall and exterior walls Total thickness (pm) F0 / F2nd mode (Out of plane) SMR 200 / 33 8 / 3 5 / 6 3.4232 116.12kHz 727.7656kHz NEMS 19.5 / 0.5 0.2 725.67kHz / 4.548 MHz

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091] For example, the SMR 1 resonator has a single embedded beam 11, a channel 3 pm deep, 8 pm wide, a central wall 5 pm wide, and external walls 6 pm wide, for a total width of 33 pm. The beam 11 of the SMR resonator has a length of 200pm, and the thickness of the lower and upper membranes covering the buried channel is 200nm. Taking into account the bonding interface oxide layer for sealing the top cover, the total thickness of the SMR resonator is estimated to be 3.4232pm. This set of dimensional parameters leads to an estimated theoretical resonance frequency of around 116.12kHz for its fundamental out-of-plane bending mode, and 727.766kHz for its second bending mode. Assuming an Allan variance of the order of ppm (106), a mass detection limit of 14.4fg is estimated. Considering a beam 21 of a NEMS sensor 2 which is mechanically coupled to the SMR resonator, the coupling is carried out for example by means of one or more suspended nano-beams 3 located perpendicular to the longitudinal axis of the beam of the SMR resonator, at the level of the first anti-node of the second bending mode of the SMR resonator from its embedding ([Fig.4A]). For example, it is assumed that the NEMS sensor and its nanobeams have a thickness equivalent to that of the upper cover covering the fluidic channel of the SMR resonator. The width and length of the beam 21 of the NEMS sensor 2 are dimensioned so that its resonance frequency according to its fundamental out-of-plane natural mode has a value similar, if not close to the frequency of the second bending mode of the SMR resonator. For the sizing of the NEMS 2 sensor, in relation to the working frequency of the SMR, we rely on the following analytical formula resulting from the application of the Euler-Bernouilli theorem: £ [ËT J n 2æ y pS Where E denotes the Young's modulus, S corresponds to the section of the NEMS beam in oscillation, I its moment of inertia according to the direction of vibrations. Xn is an eigenvalue determined according to the order of the mode, its type (bending, torsion, etc.) and the boundary conditions of the beam obtained by solving the differential equation of the beam's motion recalled below, in the absence of damping and external forces actuating it: xf) ædxrj EI-^+pS-^^O 3 xdt In the case where we consider bending in an out-of-plane direction: f -tâ [He J n 4;~ y 3p With :

[0092] / =

[0093] and w and t which respectively denote the width and thickness of the NEMS sensor beam.

[0094] The following table lists the values ​​of Xn for single-embedded and double-embedded beams: A! A2 A3 a4 Single recessed 1.875 4.694 7.854 10.995 Double recessed 4.73 7.853 10.995 14.14

[0095] Given the equation, we can deduce the expression of the fundamental resonance mode of a double-fixed NEMS beam given by the following formula:

[0096] fz 4,732 HT '1 L2

[0097] Therefore:

[0098] L = 4.73^~j~

[0099] And for a simple embedded beam, we obtain:

[0100] L=1 875 / ZÇ-JÇ"

[0101] For a thickness of 200nm of the beam 21 of the NEMS sensor 2, if we consider for example a length of 19.5qm and a width of 0.5qm, we estimate a fundamental natural frequency value of 725.67kHz, i.e. an error of 0.2% compared to the estimated value of the second out-of-plane bending resonance mode of the SMR resonator. If now the length of the beam of the NEMS sensor is 19.48qm, we then estimate a fundamental natural frequency value of 727.161kHz, i.e. an error reduced to 0.08% compared to the estimated value of the second mode of the SMR resonator 1. On the other hand, controlling the length of the beam 21 of the NEMS sensor 2 with a dimensional resolution of the order of 10nm appears incompatible to date with the precision of nano-fabrication processes combining lithography and nano-etching. An Allan variance of the order of ppm (106) is assumed for the NEMS sensor.

[0102] For its fundamental out-of-plane bending mode, a mass detection limit of the NEMS sensor is estimated at 2.14 ag, i.e. a gain of 6728 compared to the detection limit of the SMR resonator.

[0103] As expected, the reduction in size of the NEMS 2 sensor is therefore favorable to the improvement (here of three orders of magnitude) of the mass detection limit compared to the SMR 1 resonator alone. Applications

[0104] Listed below are some examples of possible applications of the system of the invention, taking advantage of the versatility offered by the increased section of the fluidic channel 12 of the SMR resonator 1 compared to its nanometric version (SNR), combined with the mass measurement sensitivity of the NEMS sensor 2. - Weigh polydisperse samples, containing particles (whether inorganic, biological, synthetic or a mixture of these different objects) with size scales ranging from a few nanometers to a few micrometers (given the section of the fluidic channel of the SMR resonator which is approximately 20x20qm2). - Weighing complex heterogeneous biological samples, for example blood, or other biofluids, in which coexist for example cells (red blood cells, lymphocytes, circulating tumor cells), circulating DNA, circulating RNA, proteins, cellular debris, extracellular vesicles (EVs), exosomes, potentially viruses or even bacteria. This set has size ranges from nm to several pm, and floating masses ranging from 0.1 attogram to several picograms. - Isolate a biological cell in the fluidic channel of the SMR resonator, by trapping it in a hydrodynamic trap (see variant of [Fig.l 1]). Given the mass resolution of the NEMS 2 sensor, it would be possible to measure with an accuracy of the order of the attogram, the rate of increase or loss of mass of the cell following either its growth or cell death or its reaction in the broad sense to the exposure of a molecule (drugs). Another characteristic of this "in-situ" trapping, combined with the mass resolution of the NEMS sensor concerns the dynamic monitoring of cellular transfection by targets such as viral nanoparticles (adenoviruses for example) or conversely the production and secretion of viral nanoparticles or extracellular vesicles (EVs), objects characterized by a floating mass ranging from a few attograms to a few hundred attograms.This real-time secretion tracking capability can also be implemented by trapping the cell upstream of the fluidic channel integrated into the SMR resonator, which allows the production kinetics of secretions flowing into the fluidic channel of the SMR resonator to be clearly monitored, without possible bias with the intrinsic mass increase rate of the cell. - Another example concerns the highlighting by weighing of target biomarkers, either by grafting to the previously functionalized surface of the fluidic channel integrated into the SMR resonator, or by grafting onto one or several beads trapped in the SMR resonator by means of hydrodynamic traps. Again, the advantage of this invention over SNR technology is that the trapped bead can be much larger, several micrometers in diameter, its developed surface area allowing it to accommodate a much larger number of probes than a sub-micrometer bead trapped in an SNR would allow. Functioning

[0105] We consider a system according to the particular embodiment described above and which comprises: - A mechanical resonator 1 of the SMR type; - A NEMS type 2 gravimetric sensor; - A connecting element 3 mechanically coupling the suspended part of the SMR resonator to the NEMS gravimetric sensor;

[0106] To calibrate the mass sensitivity of the NEMS 2 sensor with respect to the mass variations resulting from the passage of particles in the SMR resonator, a monodisperse population of reference nanoparticles (i.e. of a priori known density and size), such as gold nanoparticles, is circulated in the fluidic channel of the SMR resonator. The passage of each nanoparticle in the fluidic channel 12 of the SMR resonator 1 results in a transient modification of its effective mass meff, and therefore a frequency shift which is propagated in the suspended part 21 of the NEMS 2 sensor which is mechanically coupled to it. The frequency shift AF_nems measured by the NEMS 2 sensor must a priori be maximum when the particle is positioned at a location in the fluidic channel of the SMR resonator where the mechanical coupling between the SMR resonator and the NEMS sensor is optimal.

[0107] In the invention, as indicated above, this is a position in which the nanoparticle is located on a vibration maximum (anti-node) of the SMR resonator 1.

[0108] By measuring the frequency shift AF_nems measured using the NEMS 2 sensor, a sensitivity expressed in grams per Hertz is deduced. Once this sensitivity is calibrated, it is then possible to inject a sample into the SMR 1 resonator and deduce the mass of the particles circulating there, without first knowing their density.

[0109] The advantage of this invention lies in the fact that the dynamic range of size of the particles and targets circulating in the fluidic channel 12 of the SMR resonator 1 is extended (compared to SNR-only type technologies).

[0110] The mechanical coupling of the NEMS sensor 2 with the SMR resonator 1 is advantageously designed so that the mechanical connection is made at a vibration antinode (also called anti-node) of the SMR resonator 1 on the vibration mode for which the latter is actuated in order to maximize the transduction between it and the NEMS sensor.

[0111] The particles P of interest are conveyed into the fluidic channel 12 of the SMR type mechanical resonator 1. Their circulation in the SMR resonator 1 causes a transient AF_smr shift in the resonance frequency of the SMR resonator 1 as is the case for conventional SMR resonators (taken alone, i.e. without mechanical coupling).

[0112] On the other hand, according to the invention, the reading of this frequency variation is carried out by means of the frequency tracking of the NEMS sensor 2 which is mechanically coupled to the SMR resonator. As the NEMS sensor has a suspended part 21 having an effective mass (fraction of the total mass of the NEMS, and which depends on the requested vibration mode) reduced in comparison with the effective mass of the suspended part 11 of the SMR resonator, its detection limit is in theory much lower, even with a frequency stability of the order of ppm. It is therefore easier to read the frequency variation AF_nems at the level of the NEMS sensor 2. Manufacturing

[0113] - [Fig.12]

[0114] The system of the invention can be manufactured using techniques already used for the manufacture of SMR and SNR solutions.

[0115] This involves, for example, using two SOI ("Silicon On Insulator") substrates bonded to each other (front face to front face) by molecular sealing, according to the following arrangement: - A lower SOI substrate (denoted SOI_inf in [Fig.12]) whose active silicon layer (typically around 20qm thick for an SMR) is partially etched in order to delimit the shape of the fluidic channel which is integrated into the SMR resonator, - An upper SOI substrate (denoted SOI_sup in [Fig. 12]) whose active silicon layer is sealed by direct molecular sealing on the upper face of the lower SOI substrate.

[0116] The main manufacturing steps are, for example, the following:

[0117] El: Etching of the fluidic channel 12 in the lower SOI substrate as is customary for SMR or SNR technology;

[0118] Lithography and dry etching to etch the active silicon layer of the lower SOI substrate over its entire thickness. This step makes it possible to define one or more holes / cavities adjacent to the imprint of the fluidic channel in the SMR resonator. Chemical etching of the buried oxide layer (called BOX = buried oxide) exposed through these holes. This chemical etching leads in particular to the suspension of what will constitute the suspended part 11 of the SMR resonator 1 or a fraction of its surface. Let us specify that at least one or more holes are positioned directly above the future suspended part of the NEMS 2 sensor and its nanobeams for connection with the SMR resonator which will be etched later.

[0119] E2: Once the holes or cavities have been made, molecular sealing is then carried out on the front face of this stack with the upper SOI substrate, the thickness of the active layer of which corresponds to the thickness of the future NEMS 2 sensor and its nanobeams (=connecting element 3).

[0120] Thinning of the upper SOI substrate. The residual active layer of the upper SOI substrate is therefore used to seal the fluidic channel integrated into the future SMR resonator, and also serves as a structural material in which the shape of the NEMS 2 sensor will subsequently be delimited, as well as its mechanical connection elements to the SMR resonator and possible nano-strain gauges for piezoresistive measurement.

[0121] E3: Etching of the outline of the suspended part 11 of the SMR resonator 1, of the suspended part 21 of the NEMS sensor, its mechanical coupling element(s) with the SMR, and all other patterns necessary for the implementation of the component.

[0122] Other steps are then implemented to finalize the manufacture of the system, in particular sealing of a glass fluidic cover, structuring to delimit cavities necessary for operation.

[0123] Other manufacturing techniques could of course be used.

[0124] The invention thus presents numerous advantages, among which: - It exploits the small dimensions of the NEMS type gravimetric sensor and takes advantage of its low effective mass, and therefore its improved detection limit compared to that of the SMR type mechanical resonator. - By coupling the NEMS gravimetric sensor to the SMR type mechanical resonator, we use the advantage that the latter has to inject more complex samples than if they were injected into an SNR resonator (taken alone) with much smaller channel dimensions.

Claims

Claims

1. System for measuring at least one property of a particle (P), comprising: - A mechanical resonator (1) comprising at least a first fixed part (10) and a first suspended part (11) capable of vibrating relative to the first fixed part, and a fluidic circuit integrated in its suspended part, in which a fluid containing said particle (P) is caused to circulate, - Excitation means (4) configured to vibrate the first suspended part (11) at an excitation frequency (F_smr), - Characterized in that it comprises: - A first gravimetric sensor (2) comprising at least a second fixed part (20) and a second suspended part (21) capable of vibrating relative to the second fixed part (20) at a vibration frequency (F_nems),- The first suspended part (11) of the mechanical resonator (1) and the second suspended part (21) of the first gravimetric sensor (2) being mechanically coupled to each other via a mechanical connecting element (3), - Means for measuring variations in the vibration frequency of the second suspended part (21) of the first gravimetric sensor.,

2. System according to claim 1, characterized in that the excitation frequency corresponds to the resonant frequency of the resonator or that of a higher natural mode of this resonant frequency.

3. System according to claim 2, characterized in that the mechanical connection element (3) comprises a first end secured to the first suspended part (11) at a first point corresponding to a vibration antinode of the first suspended part, when the latter is excited at its resonance frequency (F_smr) or at that of a higher natural mode.

4. System according to claim 3, characterized in that the mechanical connecting element (3) comprises a second end secured to the second part suspended on a second point corresponding to a anti-vibration node of the second suspended part (21), when the latter is excited at the vibration frequency (F_nems).

5. System according to one of claims 1 to 4, characterized in that the first suspended part (11) is chosen from a beam embedded on a single fixed part (20), a beam embedded between two fixed parts (20), and a plate held between several fixed parts.

6. System according to one of claims 1 to 5, characterized in that the second suspended part (21) is chosen from a beam embedded on a single fixed part, a beam embedded between two fixed parts, and a plate held between several fixed parts.

7. System according to one of claims 1 to 6, characterized in that said excitation means (4) are configured to vibrate the second suspended part (21) at a vibration frequency close to the resonance frequency of the first suspended part (11) or that of a higher natural mode, at plus or minus 30% of said resonance frequency or that of the higher natural mode.

8. System according to one of claims 1 to 7, characterized in that the excitation means (4) comprise at least one piezoceramic element.

9. System according to one of claims 1 to 8, characterized in that the measuring means comprise one or more piezoresistive gauges (PZ_2).

10. System according to one of claims 1 to 9, characterized in that the fluidic circuit integrated into the first suspended part of the mechanical resonator comprises a main fluidic channel (12) and a hydrodynamic trap positioned in its channel and intended to trap said particle.

11. System according to one of claims 1 to 10, characterized in that it comprises: - A second gravimetric sensor (2), comprising at least a third fixed part and a third suspended part capable of vibrating relative to the third fixed part at a second vibration frequency, - The first suspended part (11) of the mechanical resonator (1) and the third suspended part of the second gravimetric sensor (2) being mechanically coupled to each other via a second mechanical connecting element (3), - Second means for measuring variations in the second vibration frequency of the third suspended part of the second gravimetric sensor.

12. System according to claim 11, characterized in that the third suspended part is chosen from a beam embedded on a single fixed part, a beam embedded between two fixed parts, and a plate held between several fixed parts.

13. Method for measuring at least one property of a particle, implemented using a measuring system as defined in one of claims 1 to 12, characterized in that it comprises the following steps - Injection of a fluid containing said particle (P) into the fluid circuit of the mechanical resonator (1), - Excitation of the first suspended part (11) of the mechanical resonator (1) at its resonance frequency, - Excitation of the second suspended part (21) of the first gravimetric sensor (2) at the vibration frequency, - Measurement of the variations in the vibration frequency of the second suspended part (21) of the first gravimetric sensor.

14. Measuring method according to claim 13, characterized in that it comprises a step of calibrating the system by injecting into the fluid circuit a fluid containing one or more calibrated particles.

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