Automated system for preparing a biological sample

Abstract

The invention relates to an automated system for preparing a biological sample (X) containing biological species (Y), comprising: A support plate (M0) in which one or more through-wells (10) are formed; Each through-well (10) having two opposing accesses, a first access and a second access, the first access being separated from the second access by a porous wall (100), forming a filter, against which the biological sample to be lysed can be deposited; And for each well: A first movable element (20), actuable in translation along the axis of the well to insert itself through its first access; A second movable element (30), actuable in translation along the axis of the well to insert itself through the second access; Mechanical and / or thermal stimulation means (4) for each first movable element and / or each second movable element and / or the support plate. Figure to be published with the abbreviation: Figure 1

Description

Title of the invention: Automated system for preparing a biological sample 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. The lysis of biological cells is often necessary to recover and study intracellular material, for example the cell's DNA.

[0003] The methods conventionally used for the lysis of biological species are as follows:

[0004] - Chemical lysis which consists of adding a chemical lysis solution to biological species to burst the cells; - Thermal shock lysis which consists of subjecting biological species to a temperature cycle; - Lysis by electric field which consists of subjecting biological species to an electric field; - Mechanical lysis which consists of grinding the biological species to release the biological material;

[0005] 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.

[0006] These prior solutions are however not suitable for carrying out lyses of biological samples at a high rate, and possibly with several samples in parallel.

[0007] 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

[0008] This aim is achieved by an automated system for preparing a biological sample containing biological species, comprising:

[0009] - 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 said biological sample to be lysed can be placed, - And for each well: • A first mobile member, operable in translation in the axis of the well to insert itself through its first access, • A second mobile member, operable in translation in the axis of the well to insert itself through the second access, - Means for driving each first moving member and each second moving member in translation, - Means of mechanical and / or thermal stimulation of each first mobile member and / or each second mobile member and / or the support plate, - A control unit configured to: • Order said training means, • Order said means of stimulation.

[0010] 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 awaiting use and a working position in which it is positioned to receive a biological sample in each well.

[0011] 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.

[0012] According to another particularity, the mechanical type stimulation means comprise vibratory type actuation means cooperating with the first mobile member and / or the second mobile member.

[0013] According to another particularity, the mechanical type stimulation means comprise means for actuating in rotation around its axis the first mobile member and / or the second mobile member.

[0014] According to another particularity, the thermal type stimulation means comprise a heating element integrated in the first mobile member and / or the second mobile member.

[0015] According to another feature, the first mobile member and / or the second mobile member has a free end and in that the system comprises for the first

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[0024] movable member and / or the second movable member a protective cap (90) arranged on its free end. Another feature of the system is that it includes a module that combines several protective caps. 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. The invention also relates to a method for preparing a biological sample containing biological species, implemented 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 for inserting the first movable member in translation into each well through its first access until it comes to bear against said porous wall, - Control of the drive means for inserting the second movable member in translation into each well through its second access until it comes to bear against said porous wall, - Control of the mechanical and / or thermal stimulation means of the first mobile member and / or the second mobile member associated with each well and / or the support plate to lyse the biological species. Brief description of the figures Other characteristics and advantages will appear in the detailed description which follows, given with reference to the attached drawings in which: - [Fig.l] shows the simplified architecture of the system of the invention; - [Fig.2] represents an improved version of the system of the invention; - [Fig.3] shows a particular architecture of the system of the invention; - [Fig.4] illustrates the different stages of the preparation process implemented work using the system according to the invention; Detailed description of at least one embodiment In the remainder of the description, the terms "top", "bottom", "upper", "lower" are to be understood taking into account an axis (A) drawn vertically on the attached 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.

[0025] The system is presented for example in the form of a device for lysing biological samples which can operate completely autonomously.

[0026] 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.

[0027] The control unit may comprise a programmable controller equipped with a central unit module and several input / output modules.

[0028] The system of the invention comprises a support plate MO.

[0029] In the device, the support plate MO is advantageously positioned horizontally.

[0030] The support plate MO comprises one or more through wells 10, made through its thickness. In the attached figures, the support plate MO comprises several wells but it should be understood that the principle of the invention applies to a single well of the plate.

[0031] Each well 10 of the MO plate comprises a first access, called the upper access, and a second access, called the lower access.

[0032] Between its upper access and its lower access, a well 10 comprises a transverse porous wall 100, forming a filter, between its upper part and its lower part.

[0033] The wall 100 may have a smooth or rough surface on the upper side and / or on the lower side. The biological species Y to be lysed are placed against the rough wall, to facilitate and accelerate the grinding of its biological species.

[0034] With reference to [Fig. 1], for each well 10 of the support plate MO, the system comprises:

[0035] - 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 insert itself through its lower access, - Means of mechanical and / or thermal stimulation of the first mobile member and / or the second mobile member.

[0036] 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.

[0037] The drive means 5 of the system are in particular configured and arranged to:

[0038] - Move the first movable member 20 and the second movable member 30, in translational movements (see below), - Move the MO support plate and replace it, - Move each module of the system, according to the step of the process that is implemented.

[0039] It should indeed be noted that the system may comprise several support plates MO, for example all identical, present in a storage space and waiting to be put into 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.

[0040] 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.

[0041] These dispensing means are optional because the MO support plate could be supplied pre-filled with a sample X to be treated present in each well.

[0042] For all the wells of the support plate, the dispensing means can be combined into a single dispensing module M1 comprising several reservoirs 70.

[0043] The drive means 5 are configured to place the dispensing module M1 above the support plate MO, by positioning each reservoir opposite a separate well 10 to deliver a sample X into each well.

[0044] Once the samples X are injected into each well 10 of the support plate MO, the dispensing module M1 is removed, by control of the drive means 5.

[0045] 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.

[0046] The first movable member 20 is inserted in translation into the well via its upper access and the second movable member 30 is inserted in translation into the well via its lower access. The transverse wall 100 of the well 10 can thus be clamped between the two movable members.

[0047] 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.

[0048] The free end of the mobile member intended to lyse the biological species may or may not be structured, to facilitate the grinding operation.

[0049] All of the first mobile members can be combined into a single module M2 which can be activated for all the wells of the support plate.

[0050] Similarly, all of the second moving members can be combined into a single module M3 that can be operated for all the wells of the support plate.

[0051] 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:

[0052] - The first movable member comes to bear by 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.

[0053] The two movable members make it possible to create a mechanically stable solution, in which the lower movable member creates support for the upper movable member during lysis.

[0054] For each well, the system comprises means for stimulating the first mobile member and / or the second mobile member. These stimulation means are intended to allow the lysis of the biological species present in the biological sample.

[0055] In a non-limiting manner, the stimulation means 4 may be of the mechanical and / or thermal type.

[0056] The mechanical stimulation means 4 may be of the vibratory type. In this case, the vibration may be applied to the mobile member intended to be in contact with the biological species Y to be lysed. It is also possible to maintain the two mobile members in a fixed position and to put the support plate into vibration.

[0057] It is also possible to actuate the mobile member in contact with the biological species to give it a rotational movement around a ball joint of its first end.

[0058] The thermal stimulation means 4 consist, for example, of heating the free end of the mobile member coming into contact with the biological species. The system may comprise a heating element common to the lysis module, the heat diffusing to the free end of each mobile member of the mobile. It is also possible to equip each mobile member with an independent and controllable heating element, for example a mini-heating resistor, a Peltier effect module.

[0059] 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.

[0060] 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 bringing together several elution liquid reservoirs in the same structure and a module M5 bringing together 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 MO. 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 M4 module also includes a valve system controlled by the control unit and intended to control the release of the elution liquid stored in each reservoir of the M4 module, to each well of the support plate.

[0061] For each mobile member intended to come into contact with the biological species, the system may comprise a protective cap 90 to be fitted onto 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.

[0062] [Fig. 3] schematically shows the different modules MO to M6 of the invention, showing that they can be actuated by the drive means 5, via commands from the control unit UC.

[0063] With reference to [Fig.4], the system can operate in the following manner:

[0064] - El: The control unit UC selects a support plate MO 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 commands the dispensing module Ml to deliver a biological sample X into each well of the support plate MO and pass it through the porous wall of each well. The filtered liquid is evacuated. - The control unit controls the drive means 5 to remove the dispensing module. - The control unit controls the drive means 5 to position 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 organ mobile 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 come to 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, stimulation can be of different types, mechanical and / or thermal. - Once the lysis is complete, the control unit commands 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 commands the injection of an elution liquid into each well of the support plate. The liquid containing the biological material Z of interest is collected in each receptacle of the M5 module.

[0065] The system of the invention has numerous advantages, including:

[0066] - It allows the lysis of one or more samples to be carried out in parallel in a fully automated manner; - It uses equipment conventionally used in the field of handling biological samples;

[0067] It allows lysis, without any human intervention and advantageously using single-use equipment, limiting the risks of contamination.

Claims

Claims

1. Automated system for preparing a biological sample (X) containing biological species (Y), characterized in that it comprises: - A support plate (MO) 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 porous wall (100), forming a filter, against which said biological sample to be lysed can be deposited, - And for each well: • A first mobile member (20), actuable in translation in the axis of the well to be inserted through its first access, • A second mobile member (30), actuable in translation in the axis of the well to be inserted through the second access, - Means (5) for driving in translation each first mobile member and each second mobile member,- Stimulation means (4) of the mechanical and / or thermal type for each first mobile member and / or each second mobile member and / or the support plate, - A control unit (UC) configured to: • Control said drive means, • Control said stimulation means.,

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

3. System according to claim 1 or 2, characterized in that it comprises, for each well (10) of the support plate (MO), 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 are controllable to 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 vibratory type actuation means cooperating 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 means for actuating in rotation around its axis the first mobile member and / or the second mobile member.

6. System according to one of claims 1 to 5, characterized in that the thermal type stimulation means (4) comprise a heating element integrated in 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 movable member (20) and / or the second movable member (30) has a free end and in that the system comprises for the first movable member and / or the second movable member a protective cap (90) arranged on its free end.

8. System according to claim 7, characterized in that it comprises a module (M6) bringing together several 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 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 as defined in one of claims 1 to 9, characterized in that it comprises steps of: - Control of the drive means (5) to position the support plate (MO) in a working position, - Control of the drive means (5) to insert in translation the first movable member (20) into each well through its first access until it comes to bear against said porous wall, Control of the drive means for inserting in translation the second mobile member (30) into each well through its second access until it comes to bear against said porous wall, Control of the stimulation means (4) of mechanical and / or thermal type of the first mobile member and / or of the second mobile member associated with each well and / or of the support plate (MO) to lyse the biological species (Y).

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