• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The invention relates to a device (1) for segmentation of DNA samples (A) dissolved in a container (C), the device (1) comprising: - a tank (2) intended to receive a liquid (L) ), the tank (2) being equipped with an ultrasonic wave generating means (3) for propagating ultrasonic waves in the liquid (L); and - a first support element (4) resting on the tank (2), the device (1) being characterized in that it further comprises a second support element (5) comprising a through orifice (50) intended to receive the container (C), the second support member (5) being suspended by at least one suspension element (6) forming at least one ball joint so as to allow the immersion of a lower portion of the container (C) in the liquid (L).
    公开号:FR3050211A1
    申请号:FR1653466
    申请日:2016-04-19
    公开日:2017-10-20
    发明作者:Mehdi Alizadeh；Cyrus Mahdjoubi
    申请人:Mehdi Alizadeh；Cyrus Mahdjoubi；
    IPC主号:
    专利说明:

    FIELD OF THE INVENTION The invention relates to the field of DNA segmentation which consists in obtaining, from a strand of DNA comprising a large number of nucleotide bases. , several strand portions having a smaller number of nucleotide bases. The invention more particularly relates to a device for segmenting DNA samples. 2. Prior Art
    Segmentation devices for DNA samples are known, the samples being dissolved in a dedicated container. Such devices conventionally comprise a tank intended to receive a liquid, the tank being equipped with an ultrasonic wave generating means for propagating ultrasonic waves in the liquid; and a support member resting on the tank for receiving the container. The ultrasonic wave generating means is adapted to generate an ultrasonic wave field with predefined properties in a restricted space of the vessel. This feature limits the number of DNA samples that can be simultaneously segmented in the device. A segmentation process lasts an average of one minute. Currently, the maximum number of samples segmented simultaneously by such a device is of the order of 8. However, the samples are conventionally stored in platelets comprising one hundred wells to be able to contain a hundred samples. From a productivity perspective, it would be interesting to segment all samples of a wafer simultaneously.
    In addition, DNA sample segmentation devices often require the use of specific containers, particularly when the container plays a role in the generation or transmission of ultrasonic waves. This requires a transfer of samples from an original container to a specific container, which is a time-consuming manipulation. Moreover, these specific containers represent an additional cost for the user. 3. Objectives of the invention The invention proposes a solution to overcome the aforementioned drawbacks while ensuring a device at least as powerful as the existing one with a low production cost, in particular the duration of the segmentation process is maintained at approximately one minute and the homogeneity of the length of the DNA strands obtained following the segmentation is equivalent to the existing one. The invention proposes a solution solving the technical problem of creating a substantially homogeneous ultrasound field in a volume containing a large number of DNA samples without the use of a specific container. An object of the invention is to allow the use of standard containers in the DNA sample segmentation device and in particular the use of platelets capable of containing one hundred or more samples, for the purpose of increase the productivity of such a device. 4. SUMMARY OF THE INVENTION The invention relates to a device for segmentation of DNA samples dissolved in a container, the device comprising a tank intended to receive a liquid, the tank being equipped with a means for generating ultrasonic waves for propagating ultrasonic waves in the liquid; and a first support element resting on the vessel, the device being characterized in that it further comprises a second support element comprising a through orifice intended to receive the container, the second support element being suspended from said first support element at a distance said first support member by at least one suspension member forming at least one ball joint so as to allow the immersion of a lower portion of the container in the liquid. The term "ball joint" means a link completely linking two parts in translation but leaving them free in rotation. Such a link therefore comprises 3 degrees of translation links and 3 degrees of freedom in rotation. The suspension element forming a ball joint makes it possible to substantially attenuate the transmission of parasitic ultrasonic waves from the first support element to the second support element by virtue of a damping effect of the ball joint connection. More generally, the device is designed to inhibit as much as possible the transmission of parasitic ultrasonic waves to the container. The elimination of the parasitic ultrasonic waves contributes to creating a substantially homogeneous ultrasonic wave field in the vicinity of the container. In addition, the container is not imposed by the device and various forms of container are suitable. According to an advantageous embodiment, the container is a wafer comprising a plurality of wells. The term plate or wafer is used interchangeably. Preferably, this wafer is a standard wafer commonly used to transport and / or process DNA samples.
    According to a particular embodiment, the suspension element is deformable in order to limit the propagation of parasitic waves between the first support element and the second support element.
    According to a particular embodiment, the suspension element comprises adjustment means arranged to change the distance between the first support member and the second support member. The adjustment means makes it possible to adjust the horizontality of the second support element and therefore of the container and to adjust the position of the second support element relative to the bottom of the vessel and thus the position of the container relative to the liquid.
    According to a particular embodiment, the ultrasonic wave generating means comprises at least one ultrasonic transducer transforming an electrical signal into a mechanical signal. Preferably, the ultrasonic wave generating means comprises a plurality of transducers disposed in a grid to create a relatively homogeneous ultrasonic wave field in the vicinity of the container.
    According to a particular embodiment, the ultrasonic wave generating means further comprises a signal generator arranged to supply the transducer with an electrical signal.
    According to a particular embodiment, the signal generator comprises a periodic signal generation module. For example, the signal generator comprises at least one piezoelectric oscillator.
    According to a particular embodiment, the signal generator further comprises a signal amplifier for amplifying the signal generated by the periodic signal generation module before transmitting the signal to the transducer.
    According to a particular embodiment, the signal generator further comprises a frequency and / or phase scanning module so as to generate at least two signals having distinct frequencies and / or phases in a predetermined frequency band. The scanning frequency and / or phase avoids the appearance of bellies and fixed nodes steady state in the presence of periodic waves.
    According to a particular embodiment, the ultrasonic wave generating means is fixed on an outer lower face of the tank. For example, the transducers are glued to the bottom of the tank by means of an adhesive adapted to the transmission of ultrasonic waves or are welded to the tank. The ultrasound wave generating means transmits the ultrasonic waves to the lower face of the vessel, and the lower face of the vessel in turn transmits the ultrasonic waves to the liquid.
    According to a particular embodiment, when the device is in the operating position, the ultrasonic wave generating means is placed substantially vertically to the orifice of the second support member so as to be centered with respect to the orifice. Vertical is defined as the direction of gravity. The presence of the ultrasonic wave generating means vertically to the orifice of the second support element and therefore to the vertical of the container contributes to creating a homogeneous ultrasonic wave field in the vicinity of the container by symmetry effect. However, symmetry is optional.
    According to a particular embodiment, the device comprises a second ultrasonic wave generating means for reducing or canceling the parasitic ultrasonic waves in an active manner.
    According to a particular embodiment, the first support element comprises a through hole so as to allow a user to access the orifice of the second support element. Access to the orifice of the second support member makes it possible to place a container in the second support element or to withdraw it.
    According to a particular embodiment, the suspension element comprises at least three chains in ball-and-socket connection with the upper plate and in ball-and-socket connection with the support. The three chains are distributed so as to maintain the second support member in a horizontal position while ensuring the degrees of mobility of a ball joint. The horizontality is defined as being perpendicular to the vertical direction.
    According to a particular embodiment, the second support element is remote from the lateral faces of the tank by a predetermined distance, for example a distance of 10 centimeters, so as to limit the exposure of the container to edge effects in the vicinity of the faces. lateral created by the reflection of ultrasonic waves by the lateral faces. This characteristic contributes to the homogeneity of the ultrasonic wave field in the vicinity of the container.
    According to a particular embodiment, the device further comprises an outer frame in which the bowl is placed. The frame supports the tank and isolate the external environment ultrasonic waves propagating in the tank.
    According to a particular embodiment, the device comprises a cover. The lid makes it possible to isolate the contents of the tank from the external environment for the sake of safety and hygiene.
    According to a particular embodiment, the device further comprises an overflow device intended to ensure the presence of a predetermined level of liquid in the tank. It is necessary that the liquid level is high enough to immerse the portion of the container comprising the DNA samples to be segmented. However, it is preferable that the second support element is not immersed in order to avoid the edge effects related to the reflection of the ultrasonic waves on the second support element, which would harm the homogeneity of the ultrasonic wave field.
    According to a particular embodiment, the overflow device comprises a drain orifice in order to evacuate an excess portion of the liquid.
    According to a particular embodiment, the device further comprises a means for adjusting the horizontality of the tank to ensure a homogeneous immersion of all the wells of the container in the liquid.
    According to a particular embodiment, the means for adjusting the horizontality of the tank comprises adjustable feet.
    According to a particular embodiment, the means for adjusting the horizontality of the vessel comprises a spirit level. 5. List of Figures Other features and innovative advantages will become apparent from the following description, provided for information only and by no means limitative, with reference to the accompanying drawings, in which:
    Figures la and Ib show a schematic sectional view of a device respectively according to a first and a second embodiment of the invention;
    FIG. 2 represents a schematic perspective view of the device of FIG. 1a or FIG.
    Ib; and
    FIG. 3 represents a block diagram of an ultrasonic wave generating means according to the invention. 6. Detailed description
    In Figures la, Ib and 2 is shown a device 1 for segmentation of DNA samples. The DNA samples A to be segmented are dissolved in a solution S and placed in a container C. For example, the segmentation device 1 makes it possible to obtain, from a genome comprising substantially 3 billion nucleotide bases, several strand portions having substantially 400 bases. The solution S used is typically water or an aqueous gel. The container C is preferably a standard container, for example a wafer having a plurality of wells arranged according to a grid, or a row of test tubes.
    The device 1 comprises a tank 2, an ultrasonic wave generating means 3, a first support element 4, a second support element 5, a suspension element 6, a chassis 7, a cover 8 and a control means horizontality 9 of the tank.
    The tank 2 is a sealed container designed to receive a liquid L, typically water. The tank 2 is advantageously metallic and more particularly stainless steel so as to prevent the oxidation of the tank 2 in contact with water and to facilitate its cleaning for the sake of hygiene. The tank 2 has for example a parallelepipedal shape and has a lower face and four lateral faces. From a dimensional point of view and purely illustrative, the lower face is substantially rectangular in shape and its sides are between 25 and 35 centimeters. The lateral faces have a vertical dimension of about ten centimeters, the vertical being defined by the direction of gravity.
    The tank 2 is placed in the frame 7. The frame is a container capable of isolating the external environment of ultrasonic waves propagating in the tank 2. The frame 7 is advantageously metallic and more particularly stainless steel. The frame 7 has for example a parallelepipedal shape and has a lower face and four lateral faces. From a dimensional point of view and purely illustrative, the lower face is substantially rectangular and its sides are about forty centimeters. The lateral faces have a vertical dimension of about twenty centimeters.
    The lateral faces of the tank 2 comprise upper edges resting on the lateral faces of the frame 7 so that a space is present between the outer lower face of the tank 2 and the inner lower face of the frame 7. Advantageously, the interfaces between the upper edges of the tank 2 and the lateral faces of the frame 7 comprise seals 10 limiting the propagation of parasitic waves between the tank 2 and the frame 7.
    The first support element 4 has for example substantially a plate shape with flanges. From a dimensional point of view and purely illustrative, the plate is substantially rectangular in shape and its sides are between 25 and 35 centimeters. The first support element 4 snaps removably onto the flanges of the lateral faces of the vessel so as to guarantee a substantially fixed position of the first support element 4 with respect to the vessel 2. Advantageously, the interlocking of the first support element 4 with the tank has a game for example filled by damping joints 10 limiting the propagation of parasitic waves between the tank 2 and the first support member 4. The first support member 4 has a through hole 40 substantially in its center. From a dimensional point of view and purely illustrative, the through hole 40 is substantially rectangular in shape and its sides are about fifteen centimeters. The first support element 4 is advantageously metallic and more particularly stainless steel so as to prevent its oxidation on contact with water and to facilitate cleaning. The first support element 4 advantageously comprises handles 41 facilitating its handling. For example, the handles 41 are sets of two orifices in the first support element arranged to make it possible to grip the first support element by respectively introducing two fingers into the orifices so as to grip the first support element 4.
    The second support element 5 has for example substantially a plate shape. From a dimensional point of view and purely illustrative, the plate is substantially rectangular in shape and its sides are about fifteen centimeters. The second support member 5 has a through hole 50 substantially at its center. From a dimensional point of view and purely illustrative, the through hole 50 is substantially rectangular in shape and its sides are about ten centimeters. The second support element 5 is advantageously metallic and more particularly stainless steel so as to prevent its oxidation on contact with water and to facilitate cleaning.
    The second support element 5 is suspended from the first support element 4 by the suspension element 6 forming a ball joint. The suspension element 6 advantageously comprises 4 chains 61 respectively connecting four points of the support element 4, near the four corners of the orifice 40, at four points of the support element 5, near the four corners of the support 5. Each chain 61 has two ends, the first end being in ball-and-socket connection with the first support element 4 and the second end being in ball-and-socket connection with the second support element 5. According to other embodiments, the chains 61 are replaced. by stems, rings or ropes. Such deformable suspension elements 6 extend between the first support element 4 and the second support element 5 along a length for example of approximately 4 centimeters and make it possible to keep the second support element substantially in the center of the vessel in a horizontal position, the horizontal being defined as perpendicular to the vertical, while limiting the transmission of ultrasonic waves through the suspension element 6 through the damping character conferred by the ball joint.
    According to a particular embodiment represented in FIG. 1b, the suspension element 6 comprises a setting means arranged to modify the distance between the first support element 4 and the second support element 5. The adjustment means comprises two sleepers 63 substantially perpendicular to the chains 61 and four screws 62 connecting the first support member 4 to the cross members 63. By screwing or unscrewing the screws 62, the distance between the first support member 4 and the cross members 63 is changed. Advantageously, the four screws 62 are inserted into the first support element 4 respectively at four points of the support element 4 near the four corners of the orifice 40. Two adjacent screws are screwed respectively into the two ends of a first cross member. the other screws are screwed respectively into the two ends of the second crossmember. Four points of the second support element 5 near the four corners of the support element 5 are respectively connected to the four ends of the two crosspieces by chains 61 substantially in the extension of the screws 62.
    The second support member 5 is for receiving a container C, typically a wafer having a plurality of wells for receiving DNA samples. The container C is placed so that the wells are in the orifice 50 and the direction in which the wells extend is vertical, the ends of the container, at the periphery of the wells, resting on the second support element 5.
    The second support element 5 is remote from the lateral faces of the vessel by a predetermined distance. for example a distance of 10 centimeters, so as to limit the exposure of the container C to the edge effects in the vicinity of the side faces created by the reflection of the ultrasonic waves by the side faces.
    The tank 2 is filled with liquid L so as to immerse the distal end of the wells and to leave the second support element 5 in the open air in order to avoid the reflection of ultrasonic waves propagating in the liquid L by the element 5. The tank 2 comprises for example between 5 and 7 centimeters of liquid L. During the filling operation of the tank 2, the first and second support elements are removed from the tank 2 so as to pour the liquid L in the tank 2. The liquid level L is predefined and must be precise to the millimeter. In order to precisely control the liquid level L in the tank 2, the tank 2 comprises an overflow device 21. According to the embodiment shown, the overflow mechanism comprises a drain orifice 210 in a lateral face of the tank at a height corresponding to the desired liquid level. The excess liquid is discharged through a conduit 211. The conduit 211 conveys the excess liquid to the outside of the frame 7 through a through hole in the frame 7. According to an embodiment not shown, the excess liquid is collected in a container attached to the frame 7. According to another embodiment not shown, the overflow device 21 has a wall delimiting a sealed compartment in the tank 2, the wall being arranged so that the excess liquid passes over an upper edge of Wall.
    The cover 8 covers the frame 7 and allows to isolate the contents of the tank 2 of the external environment for the sake of safety and hygiene.
    Advantageously, the cover 8 has seals 10 at the interfaces between the cover 8 and the first support element 4 or between the cover 8 and the tank 2 so as to limit the propagation of parasitic waves at these interfaces.
    The means for adjusting the horizontality 9 of the tank comprises adjustable feet 91 and a spirit level 92. In the embodiment shown, the adjustable feet 91 and the spirit level 92 are fixed on the lower external face of the frame 7.
    The ultrasonic wave generating means 3 comprises a plurality of transducers 31, for example 6 transducers arranged in a grid comprising 2 lines of 3 transducers in order to create a relatively homogeneous ultrasound wave field in the vicinity of the container C. The transducers transform an electrical signal in a mechanical signal generating ultrasonic waves. The transducers are attached to an outer face of the vessel 2, for example they are glued by means of an adhesive adapted to the transmission of ultrasonic waves or they are welded. Advantageously, the transducers are fixed on an external lower face of the tank, vertically of the orifice 50 of the second support element 5. FIG. 3 shows a means for generating ultrasonic waves 3. The transducers 31 are supplied with an electrical signal by a signal generator 32. Advantageously, the signal generator 32 comprises a periodic signal generation module 322, for example a piezoelectric oscillator, and a signal conditioning and power electronics module 321 amplifying the signal generated by the periodic signal generation module 322. Advantageously, in order to create a substantially homogeneous ultrasonic wave field in the vicinity of the container and , in particular, in order to avoid the appearance of stomachs and nodes in stationary mode, the signal generator 32 comprises a frequency scanning module 323 so as to generate at least two signals having distinct frequencies in a frequency band predetermined, for example a frequency band between 24kHz and 28kHz. Advantageously, the signal generator 32 furthermore comprises an adjustable modulation module in amplitude and in phase in order to be able to vary the amplitude and the phase of the signals generated, always with the aim of avoiding the appearance of bellies and nodes. . The invention is described in the foregoing by way of example. It is understood that one skilled in the art is able to achieve different embodiments of the invention, for example by combining the various characteristics above taken alone or in combination, without departing from the scope of the invention. .
    权利要求:
    Claims (10)
    [1" id="c-fr-0001]
    1. Device (1) for segmentation of DNA samples (A) dissolved in a container (C), the device (1) comprising: a tank (2) intended to receive a liquid (L), the tank (2) being equipped with an ultrasonic wave generation means (3) for propagating ultrasonic waves in the liquid (L); and a first support member (4) resting on the tank (2), the device (1) being characterized in that it further comprises a second support member (5) having a through orifice (50) for receiving the container (C), the second support member (5) being suspended from said first support member (4) at a distance from said first support member (4) by at least one suspension member (6) forming at least one ball joint so as to allow the immersion of a lower portion of the container (C) in the liquid (L).
    [2" id="c-fr-0002]
    2. Device (1) according to claim 1 characterized in that the suspension element (6) is deformable.
    [3" id="c-fr-0003]
    3. Device (1) according to claim 1 or 2 characterized in that the suspension element (6) comprises an adjusting means (62-63) arranged to change the distance between the first support member (4) and the second support element (5).
    [4" id="c-fr-0004]
    4. Device (1) according to any one of claims 1 to 3, characterized in that the ultrasonic wave generating means (3) comprises at least one ultrasonic transducer (31).
    [5" id="c-fr-0005]
    5. Device (1) according to claim 4 characterized in that the means for generating ultrasonic waves (3) further comprises a signal generator (32) arranged to supply electrical signal to the transducer.
    [6" id="c-fr-0006]
    6. Device (1) according to claim 5 characterized in that the signal generator (32) comprises a periodic signal generation module (322).
    [7" id="c-fr-0007]
    7. Device (1) according to any one of the preceding claims characterized in that the ultrasonic wave generating means (3) is fixed on an outer lower face of the vessel (2).
    [8" id="c-fr-0008]
    8. Device (1) according to claim 7 characterized in that the ultrasonic wave generating means (3) is placed substantially vertically to the orifice (50) of the second support member (5).
    [9" id="c-fr-0009]
    9. Device (1) according to any one of the preceding claims characterized in that the first support member (4) comprises a hole (40) passing through so as to allow a user to access the orifice (50) of the second support member (5).
    [10" id="c-fr-0010]
    10. Device (1) according to any one of the preceding claims characterized in that the second support member (5) is remote from the side faces of the tank (2) by a predetermined distance so as to limit the exposure of the container (C) edge effects in the vicinity of the lateral faces created by the reflection of the ultrasonic waves by the lateral faces.
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    法律状态:
    2017-04-07| PLFP| Fee payment|Year of fee payment: 2 |
    2017-10-20| PLSC| Publication of the preliminary search report|Effective date: 20171020 |
    2018-02-27| PLFP| Fee payment|Year of fee payment: 3 |
    2018-03-16| TQ| Partial transmission of property|Owner name: CYRUS MAHDJOUBI, FR Effective date: 20180213 Owner name: ETABLISSEMENT FRANCAIS DU SANG, FR Effective date: 20180213 |
    2018-05-18| TP| Transmission of property|Owner name: ETABLISSEMENT FRANCAIS DU SANG, FR Effective date: 20180412 |
    2019-04-30| PLFP| Fee payment|Year of fee payment: 4 |
    2020-04-29| PLFP| Fee payment|Year of fee payment: 5 |
    2021-04-28| PLFP| Fee payment|Year of fee payment: 6 |
    优先权:
    申请号 | 申请日 | 专利标题
    FR1653466|2016-04-19|
    FR1653466A|FR3050211B1|2016-04-19|2016-04-19|DEVICE FOR SEGMENTING DNA SAMPLES|FR1653466A| FR3050211B1|2016-04-19|2016-04-19|DEVICE FOR SEGMENTING DNA SAMPLES|
    AU2017252473A| AU2017252473B2|2016-04-19|2017-06-19|Device for DNA sample fragmentation|
    ES17742477T| ES2870593T3|2016-04-19|2017-06-19|DNA sample segmentation device|
    US16/094,798| US10919018B2|2016-04-19|2017-06-19|Device for DNA sample fragmentation configured to produce ultrasonic waves|
    KR1020187033046A| KR102370478B1|2017-06-19|DNA sample segmentation device|
    PCT/FR2017/051609| WO2017182763A1|2016-04-19|2017-06-19|Device for dna sample fragmentation|
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    CA3021568A| CA3021568A1|2016-04-19|2017-06-19|Device for dna sample fragmentation|
    EP17742477.7A| EP3445493B1|2016-04-19|2017-06-19|Device for dna sample fragmentation|
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