AUTOMATED SYSTEM FOR PREPARING A BIOLOGICAL SAMPLE

Description

Technical field of the invention

[0001] The present invention relates to an automated system for preparing a biological sample. State of the art

[0002] The preparation of a biological sample often involves a step of lysis of the biological species present in the sample. Lysis of biological cells is often necessary to recover and study intracellular material, such as the cell's DNA.

[0003] The methods classically used for the lysis of biological species are as follows: Chemical lysis, which consists of adding a chemical lysis solution to the biological species to burst the cells; Thermal shock lysis, which consists of subjecting the biological species to a temperature cycle; Electric field lysis, which consists of subjecting the biological species to an electric field; Mechanical lysis, which consists of crushing the biological species to release the biological material;

[0004] Various mechanical lysis solutions have already been proposed in the state of the art, notably in patent applications WO2015 / 181743A1 and EP3222989A1 . These solutions mainly consist of grinding the biological species against a rough wall, or by shearing. Another system for preparing a sample by lysis is disclosed in US 2013 / 217113 A1.

[0005] However, these previous solutions are not suitable for performing lyses of biological samples at a high rate, and possibly with several samples in parallel.

[0006] The aim of the invention is to propose a system capable of preparing biological samples at a high rate, and possibly capable of processing several samples in parallel. Statement of the invention

[0007] This aim is achieved by an automated system for preparing a biological sample containing biological species according to claim 1, comprising: A support plate in which one or more through-wells are made, Each through-well having two opposite accesses, a first access and a second access, the first access being separated from the second access by a porous wall, forming a filter, against which the biological sample to be lysed can be deposited, And for each well: ∘ A first movable organ, actuable in translation along the axis of the well to insert itself through its first access, ∘ A second movable organ, actuable in translation along the axis of the well to insert itself through the second access, Means for driving each first movable organ and each second movable organ, Means for mechanical and / or thermal stimulation of each first movable organ and / or each second movable organ and / or the support plate, A control unit configured to: ∘ Control said driving means, ∘ Control said stimulation means.

[0008] According to one particular feature, the drive means are configured to move said support plate between at least two distinct positions, a storage position in which it is waiting to be used and a working position in which it is positioned to receive in each well, a biological sample.

[0009] According to another feature, the system comprises, for each well of the support plate, means for dispensing a biological sample into each well of the support plate, the drive means being configured to move the dispensing means between two distinct positions, a storage position and a working position in which they are controllable to inject a biological sample into each well.

[0010] According to another feature, the mechanical type stimulation means comprise vibratory type actuation means cooperating with the first mobile member and / or the second mobile member.

[0011] According to another feature, the mechanical type stimulation means comprise means for actuating the first mobile member and / or the second mobile member in rotation around its axis.

[0012] According to another particularity, thermal stimulation methods include a heating element integrated into the first moving organ and / or the second moving organ.

[0013] According to another feature, the first movable member and / or the second movable member has a free end and the system comprises, for the first movable member and / or the second movable member, a protective cap arranged on its free end. According to another feature, the system comprises a module combining several protective caps.

[0014] According to another feature, the system comprises, for each through-well, an elution device comprising a reservoir intended to receive an elution fluid to be injected into each well and a receptacle intended to recover said elution fluid, passing through the porous wall of the well.

[0015] The invention also relates to a method for preparing a biological sample containing biological species according to claim 10, carried out using an automated system as defined above, the method comprising steps of: Control of the drive means to position the support plate in a working position, Control of the drive means to insert the first moving part in translation into each well through its first access until it comes to rest against said porous wall, Control of the drive means to insert the second moving part in translation into each well through its second access until it comes to rest against said porous wall, Control of the mechanical and / or thermal stimulation means of the first moving part and / or the second moving part associated with each well and / or the support plate to lyse the biological species. Brief description of the figures

[0016] Other features and advantages will appear in the detailed description that follows, in conjunction with the attached drawings in which: There figure 1 shows the simplified architecture of the system of the invention; The figure 2represents an improved version of the system of the invention; The figure 3 shows a particular architecture of the system of the invention; The figure 4 illustrates the different stages of the preparation process implemented using the system in accordance with the invention; Detailed description of at least one embodiment

[0017] In the remainder of the description, the terms "top", "bottom", "upper", "lower" are to be understood taking into account an axis (A) drawn vertically in the appended figures. The invention relates to an automated system making it possible to carry out mechanical lysis of a biological sample at a high rate, and even to carry out several lyses simultaneously on several samples in parallel.

[0018] The system is presented, for example, in the form of a biological sample lysis device that can operate completely autonomously.

[0019] The system comprises a control unit UC responsible for controlling different modules (see below) of the system to implement the different steps of the method of the invention.

[0020] The control unit may include a programmable controller with a central unit module and several input / output modules.

[0021] The system of the invention comprises a support plate M0.

[0022] In the apparatus, the support plate M0 is advantageously positioned horizontally. The support plate M0 comprises one or more through-wells 10, made through its thickness. In the attached figures, the support plate M0 comprises several wells but it should be understood that the principle of the invention applies to a single well of the plate.

[0023] Each well 10 of the plate M0 has a first access, called the upper access, and a second access, called the lower access.

[0024] Between its upper access and its lower access, a well 10 has a transverse porous wall 100, forming a filter, between its upper and lower part.

[0025] The wall 100 can have a smooth or rough surface on the upper and / or lower side. The biological species Y to be lysed are placed against the rough wall to facilitate and accelerate the grinding of these biological species.

[0026] In reference to the figure 1 For each well 10 of the support plate M0, the system comprises: A first mobile member 20 operable in translation in the axis of the well 10 to be inserted through its upper access, A second mobile member 30, operable in translation in the axis of the well to be inserted through its lower access, Means of mechanical and / or thermal stimulation 4 of the first mobile member and / or of the second mobile member and / or of the support plate M0.

[0027] The system also comprises drive means 5 controlled by the control unit UC. These drive means 5 may comprise motors, arms, jacks or equivalent, capable of moving the modules and others of the system along various axes and in different planes.

[0028] The system's training means 5 are specifically configured and arranged to: Move the first mobile member 20 and the second mobile member 30, in translational movements (see below), Move the support plate M0 and replace it, Move each module of the system, according to the step of the method which is implemented.

[0029] It should be noted that the system may comprise several support plates M0, for example all identical, present in a storage space and waiting to be placed in the working position. The control unit UC is responsible for selecting a support plate from all the support plates available in the storage space and for controlling drive means 5 to move the selected support plate from the storage position to the working position.

[0030] The dispensing means are intended to deliver, into the well, the biological sample X to be treated. The biological sample X may be in a liquid form containing biological species Y. The liquid part may be evacuated by passing through the porous wall of the well 10, leaving only the biological species Y to be lysed on the upper face of the wall 100.

[0031] These dispensing means are optional because the M0 support plate could be supplied pre-filled with a sample X to be treated present in each well.

[0032] For all the wells of the support plate, the dispensing means can be combined into a single dispensing module M1 comprising several reservoirs 70.

[0033] The drive means 5 are configured to place the dispensing module M1 above the support plate M0, positioning each reservoir opposite a separate well 10 to deliver a sample X into each well.

[0034] Once the samples X are injected into each well 10 of the support plate M0, the dispensing module M1 is removed, by control of the drive means 5.

[0035] For each well 10, the system comprises a first mobile member 20 and a second mobile member 30, used for the lysis of the biological species Y present in the well 10.

[0036] The first movable member 20 is inserted in translation into the well through its upper access and the second movable member 30 is inserted in translation into the well through its lower access. The transverse wall 100 of the well 10 can thus be clamped between the two movable members.

[0037] The first movable member 20 and the second movable member 30 may each be in the form of a rigid rod having a first end fixed to a movable support and an opposite end which is free.

[0038] The free end of the mobile organ intended to lyse the biological species may or may not be structured, to facilitate the grinding operation.

[0039] All of the first moving parts can be combined into a single M2 module that can be operated for all the wells of the support plate.

[0040] Likewise, all the second moving parts can be combined into a single M3 module that can be operated for all the wells of the support plate.

[0041] The drive means 5 are configured to position each of the two modules M2, M3, so that their movable members are located opposite a separate well of the support plate. The drive means 5 are then configured to drive each movable member in translation so that, for each well: The first movable member rests with its free end against the upper face of the transverse wall of the well, The second movable member rests with its free end against the lower face of the transverse wall of the well.

[0042] The two movable members allow for the creation of a mechanically stable solution, in which the lower movable member creates support for the upper movable member during lysis.

[0043] For each well, the system includes means for stimulating the first moving part and / or the second moving part and / or the support plate M0. These stimulation means are intended to enable the lysis of biological species present in the biological sample.

[0044] Without limitation, the means of stimulation 4 may be of mechanical and / or thermal type.

[0045] Mechanical stimulation methods can be of the vibratory type. In this case, vibration can be applied to the moving part intended to be in contact with the biological species to be lysed. It is also possible to hold both moving parts in a fixed position and vibrate the support plate.

[0046] This activates at least one mobile organ and / or the support plate M0. It is also possible to activate everything at the same time. It should be noted that lysis can occur by relative movement. The stimulation can be applied to the mobile organ directly in contact with the biological species Y, or by stimulation of the other mobile organ, the latter cooperating with the support plate M0 to drive it in movement, or by stimulation of the support plate relative to the two fixed mobile organs.

[0047] It is also possible to operate the mobile organ in contact with the biological species to give it a rotational movement around a ball joint at its first end.

[0048] Thermal stimulation methods, for example, involve heating the free end of the mobile organ that comes into contact with the biological species. The system may include a heating element common to the lysis module, with heat diffusing to the free end of each mobile organ. Alternatively, each mobile organ can be equipped with an independent and controllable heating element, such as a miniature heating element or a Peltier module.

[0049] In these different cases, to accelerate lysis, the face of the transverse wall 100 of the well 10 supporting the biological species Y is advantageously rough.

[0050] The system may comprise means for eluting the biological material obtained in each well after lysis. For each well 10, these elution means may comprise a reservoir 80 of elution liquid intended to be placed above the well, and a receptacle 81 intended to receive the liquid after elution, located under the well. As previously, the system may comprise a module M4 combining several reservoirs of elution liquid in the same structure and a module M5 combining the receptacles. The reservoirs present in the module M4 and the receptacles present in the module M5 may be common to several wells 10 of the support plate M0. These two modules, called elution modules, are capable of being placed in position, respectively above and below the support plate during the elution of the biological material.The drive means, controlled by the control unit, are configured to position each of the two elution modules M4, M5 in their working position. The module M4 also comprises a valve system controlled by the control unit and intended to control the release of the elution liquid stored in each reservoir of the module M4, to each well of the support plate.

[0051] For each mobile member intended to come into contact with the biological species, the system may comprise a protective cap 90 to be fitted to its free end. It is possible to provide a protective module M6, combining all the caps 90 into a single element, and intended to fit onto the module M2 or M3 supporting the mobile members. The drive means are also configured to fit this protective module M6.

[0052] There figure 3schematically reproduce the different modules M0 to M6 of the invention, showing that they can be acted upon by the drive means 5, via commands from the control unit UC.

[0053] In reference to the figure 4 , the system can operate as follows: E1: The control unit UC selects a support plate M0 and controls the drive means 5 to position the selected support plate in the working position, horizontally. E2: The control unit controls the drive means 5 to position the dispensing module above the support plate. E3: The control unit controls the dispensing module M1 to deliver a biological sample X into each well of the support plate M0 and make it pass through the porous wall of each well. The filtered liquid is discharged. The control unit controls the drive means 5 to remove the dispensing module. The control unit controls the drive means 5 to place the lysis modules M2, M3. E4: The control unit controls the drive means 5 to move the lysis modules M2, M3 in translation, so that a first mobile member 20 is inserted into each well through its upper access and a second mobile member 30 is inserted into each well through its lower access. The two mobile members each bear on the two opposite faces of the transverse wall of the well. E5: The control unit controls the stimulation means 4 to lyse the biological species present in each well. As indicated above, the stimulation can be of different types, mechanical and / or thermal, applied to one or two mobile members and / or to the support plate M0.Once the lysis is complete, the control unit controls the removal of the lysis modules M2, M3. The control unit can control the elution of the biological material obtained in each well. The control unit controls the drive means to position the elution modules M4, M5, respectively above and below the support plate. E6: The control unit controls the injection of an elution liquid into each well of the support plate. The liquid containing the biological material Z of interest is recovered in each receptacle of the module M5.

[0054] The system of the invention has many advantages, including: It allows the lysis of one or more samples in parallel in a fully automated manner; It uses equipment conventionally used in the field of handling biological samples;

[0055] It allows lysis, without any human intervention and advantageously using single-use equipment, limiting the risks of contamination.

Claims

1. Automated system for preparing a biological sample (X) containing biological species (Y), characterized in that it comprises: - a support plate (M0) in which one or more through-wells (10) are made, - each through-well (10) having two opposite accesses, a first access and a second access, the first access being separated from the second access by a filter-forming porous wall (100), against which the said biological sample that is to undergo lysis may be placed, - and for each well: ∘ a first mobile member (20), that can be actuated to move translationally along the axis of the well so as to become inserted across the first access thereof, ∘ a second mobile member (30), that can be actuated to move translationally along the axis of the well so as to become inserted across the second access thereof, - drive means (5) driving the translational movement of each first mobile member and of each second mobile member, - stimulation means (4) of mechanical and / or thermal type for stimulating each first mobile member and / or each second mobile member and / or the support plate, - a control unit (UC), configured to: ∘ command said drive means, ∘ command said stimulation means.

2. System according to Claim 1, characterized in that the drive means (5) are configured to move said support plate (M0) between at least two distinct positions: a storage position in which it is on standby awaiting use and a working position in which it is positioned to receive a biological sample (X) in each well (10).

3. System according to Claim 1 or 2, characterized in that it comprises, for each well (10) of the support plate (M0), means for dispensing a biological sample (X) into each well of the support plate, the drive means (5) being configured to move the dispensing means between two distinct positions: a storage position and a working position in which they can be controlled such that they inject a biological sample into each well.

4. System according to one of Claims 1 to 3, characterized in that the mechanical-type stimulation means (4) comprise actuating means of vibratory type collaborating with the first mobile member (20) and / or the second mobile member (30).

5. System according to one of Claims 1 to 3, characterized in that the mechanical-type stimulation means (4) comprise actuating means causing the first mobile member and / or the second mobile member to rotate about its axis.

6. System according to one of Claims 1 to 5, characterized in that the thermal-type stimulation means (4) comprise a heating element incorporated into the first mobile member and / or the second mobile member.

7. System according to one of Claims 1 to 6, characterized in that the first mobile member (20) and / or the second mobile member (30) has a free end and in that the system comprises, for the first mobile member and / or the second mobile member, a protective cap (90) arranged over the free end thereof.

8. System according to Claim 7, characterized in that it comprises a module (M6) combining a plurality of protective caps (90).

9. System according to one of Claims 1 to 8, characterized in that it comprises, for each through-well (10), an elution device comprising a reservoir (80) intended to receive an elution fluid that is to be injected into each well, and a receptacle (81) intended to recover said elution fluid, passing through the porous wall (100) of the well.

10. Method for preparing a biological sample (X) containing biological species (Y), implemented using an automated system such as defined in one of Claims 1 to 9, characterized in that it comprises the steps of: - commanding the drive means (5) to position the support plate (M0) in a working position, - commanding the drive means (5) to insert the first mobile member (20) translationally into each well across the first access thereof until it comes to bear against said porous wall, - commanding the drive means to insert the second mobile member (30) translationally into each well across the second access thereof until it comes to bear against said porous wall, - commanding the mechanical-type and / or thermal-type stimulation means (4) of the first mobile member and / or second mobile member associated with each well and / or of the support plate (M0) to bring about lysis of the biological species (Y).