PRINTING OBJECTS FROM A WELL

Abstract

The present invention relates to a method and equipment for transferring an object from a donor substrate (30) to a target substrate (40), said donor substrate (30) containing the objects to be transferred of dimensions (Dx, Dy, Dz), said transfer being ensured by local energy excitation of said liquid to form a cavitation bubble localized at the level of an object, characterized in that said substrate is constituted by a plate of micro-wells having a plurality of wells each forming a receptacle having an open upper base narrowing towards the bottom, the width (L) of the base of said wells being L > 2DxDy and the height (h) of said well h > 2Dz, and the transfer is done with a first energy level E1 if the ratio between the dimension (Dz) of said object and thickness (e) of said liquid film is less than 1, and with a second energy level E2 > E1 if the ratio between the dimension (Dz) of said object and thickness (e) of said liquid film is greater than 1.Figure from the summary: figure 1.

Description

Title of the invention: PRINTING OBJECTS FROM A WELL Scope of the invention

[0001] The present invention relates to the field of additive manufacturing of a material by repeated transfers of particles between a donor substrate and a receiving substrate, a liquid carrier film containing the particles to be transferred being deposited on the donor substrate, and in particular to the field of bioprinting. The specific field of bioprinting concerns the use of digital manufacturing processes enabling the organization and assembly in 2D and 3D of the constituents of biological tissues for the purpose of producing grafts for regenerative medicine or physiological models for biomedical and pharmaceutical research.

[0002] The general principle consists of transferring biological objects (cells, for example), organic or mineral, from a carrier liquid deposited on a substrate in the form of a film, and applying an energy pulse to form a cavitation bubble, either directly by vaporizing the portion of the liquid in the focal field of the energy source—generally a laser—or via a thin metallic coating forming a sacrificial layer on the donor substrate. This cavitation bubble carries the particle(s) located along the firing axis toward a receiving substrate, on which the transferred particles accumulate as the firings are repeated. State of the art

[0003] One of the solutions suitable for bioprinting is known by the acronym LIFT (Laser-Induced Forward Transfer). The material transfer is caused by

[0004] focusing a laser onto a surface. This transfer can occur in two directions: either by ablating a surface from above and then allowing it to fall by capillary action, or by pulling the surface downwards and using the released energy to cause a material transfer to a receiver positioned above the surface. LIFT consists of projecting a bio-ink spread on a donor substrate onto a receiver substrate. The donor substrate is composed of a glass plate coated with a thin layer of gold onto which the bio-ink is spread. The receiver substrate can be, for example, a 6-well plate or a wellless plate for printing large areas.

[0005] A laser delivers short infrared pulses (from a few hundred femtoseconds down to tens of nanoseconds). The beam is magnified and directed by a set of mirrors towards a scanner. This scanner directs the beam towards a lens F- Theta, which allows it to be focused on the donor blade. The coordinates of the laser ablation points on the donor blade are controlled by the angle applied by the scanner.

[0006] When the focused laser pulse reaches the gold layer, the electrons within it absorb the laser photons. A plasma is created, and a vapor bubble forms on the glass surface, called a cavitation bubble. This bubble, located between the glass and the bio-ink, projects an ink droplet containing one or more cells onto the receiving substrate. The distance between the donor gold layer and the receiver surface is defined as the Donor-Receiver Distance (DRD). This distance is one of the main parameters influencing print quality.

[0007] For example, we know of patent EP3941712 describing a printing process implementing equipment comprising a means of orientable energy excitation to produce a point interaction with at least one ink which may contain inhomogeneities deposited on a printing support having a transparent interaction surface, in order to cause the transfer of a targeted part of said ink to a receiver, characterized in that it comprises a step of generating a wetting film covering at least partially said transparent interaction surface, followed by a step of depositing said ink on the surface of said wetting film and transfer steps.

[0008] We also know of patent application EP22181067.4. Indeed, this is a patent protecting a process for manufacturing three-dimensional cartilage tissue by laser-assisted bioprinting, comprising the following steps:

[0009] a) provide a donor bio-ink comprising cartilage-forming cell aggregates,

[0010] b) transfer a structured layer of cell aggregates onto a receiving substrate by pulsed laser energy focused on the bio-ink.

[0011] c) repeat steps a) and b) to obtain additional patterned aggregate layers

[0012] on the previously deposited layers, thus obtaining said three-dimensional layer

[0013] with a minimum compactness factor of 30%.

[0014] Among these technologies is patent EP2873751A1, which describes a process for the direct deposition of a solid material onto a target substrate by passing a burst of ultrafast laser pulses from a focused laser beam below the diffraction limit through a carrier substrate that is transparent to the laser beam. The carrier substrate is coated with the solid material to be transferred on its underside. Electrons on the back of this transparent, material-coated support are excited by the first sub-pulses of the laser beam, which lift the material from the carrier substrate and the Subsequent sub-pulses of the laser beam send the material into space at hypersonic speed by a shock wave that drives the material with forward momentum through a narrow gap between the carrier substrate and the target substrate.

[0015] The process described in this patent is based on the melting of the material to be transferred by laser absorption and the shock waves necessary for the transfer are not compatible with use in the biological field.

[0016] Kenzan bioprinting technology [Yurie et al. (2017) PLoS ONE 12(2), e0171448] is a method for assembling 3D cell aggregates without the aid of a scaffold made of collagen or hydrogel materials. Spheroids are arranged in a network of fine needles where they can fuse with adjacent spheroids to form a connected structure. By using appropriate needle alignment, the spheroids can be positioned in specific 3D arrangements, particularly for hollow constructs. This technology is limited in terms of production throughput because the spheroids must be handled individually using forceps or a syringe. Furthermore, the spheroids are generally large enough to accommodate needle insertion.Kenzan technology cannot produce dense tissues (with a high compactness factor) composed of spheroids smaller than 500 sq m in diameter. Furthermore, this technology is limited in its applications because it relies on the direct manipulation of objects. It is poorly suited to manipulating deformable objects, cells sensitive to environmental variations, or GMP conditions.

[0017] Suction-assisted bioprinting is another technique particularly suited to the manipulation and shaping of spheroids as described in the publication:

[0018] “Aspiration-assisted bioprinting for precise positioning of biologics”

[0019] Ayan B, Heo DN, Zhang Z, Dey M, Povilianskas A, Drapaca C and Ozbolat IT

[0020] 2020 Sci. Adv. 6 1-17

[0021] This technology enables the 3D harvesting and positioning of aggregates by harnessing the power of suction forces. It operates a pipette, which is used to "harvest" spheroids from a gel or bio-ink and "3D bioprint" them into or onto a gel substrate (receptor). This technology claims high-precision positioning and high viability, but the aspiration needle is in contact with the spheroid, with the potential for contamination and mechanical damage. Furthermore, to ensure good aspiration, this solution is limited to handling large spheroids and is therefore limited in terms of production throughput. As with the Kenzan technology, the aspiration technology requires handling the spheroids individually at low speeds and does not allow for This technology produces dense tissues (high compactness factor) composed of spheroids less than 500 µm in diameter. However, its applications are limited because it relies on the direct manipulation of objects. It is poorly suited to handling deformable objects, cells sensitive to environmental variations, or GMP conditions. Disadvantages of prior art

[0022] Prior art solutions are poorly suited to particles with sizes greater than 100 pm and also pose difficulties with regard to centering the shot relative to the particle to be transferred.

[0023] Moreover, prior art solutions involve object handling operations that present risks of loss of viability, particularly for fragile biological objects. Solution provided by the invention

[0024] To overcome these drawbacks, the present invention, in its most general sense, relates to a method for manufacturing a material by transferring at least one particle from a donor substrate to a target substrate. More particularly, it relates to a method for transferring an object from a donor substrate to a target substrate, a liquid carrier film containing the particles to be transferred being deposited on said donor substrate containing the objects to be transferred of dimensions (Dx, Dy, Dz), said transfer being ensured by local energy excitation to form a cavitation bubble localized at the level of an object, characterized in that a. said substrate consists of a micro-well plate having a plurality of wells, each well forming a receptacle having an open upper base narrowing towards the bottom, the width (L) of the base of said wells being L > 2DxDy and the height (h) of said well h > 2DZ, and b. the transfer takes place - According to a first transfer mode with a first energy level Ei, if the ratio between the dimension (Dz) of said object and the thickness (e) of said liquid film is less than 1 - According to a second transfer mode with a second energy level E2> Ei if the ratio between the dimension (Dz) of said object and thickness (e) of said liquid film is greater than 1.

[0025] According to a first variant, the energy deposition is achieved by focusing a laser for both transfer modes, propulsion and LIFT.

[0026] According to a second variant, energy deposition is achieved via an electric field for the propulsion mode.

[0027] According to a third variant, the energy deposition is achieved by focusing an acoustic wave for the propulsion mode.

[0028] According to a particular embodiment, the receiving substrate includes a damping means along the axis of transfer of the objects.

[0029] According to one embodiment, the transfer of biological objects is achieved without contact by the generation of a cavitation bubble in the liquid present between the object and the bottom of the well, the transfer being ensured • - either by converting the deposited energy into kinetic energy the object that lifts off and leaves the liquid layer with strong directionality in propulsion mode • - either by converting the deposited energy into liquid movement which carries objects away via a jet in LIFT mode

[0030] The contactless transfer guarantees the absence of plastic deformation of the object.

[0031] Advantageously, the objects to be transferred are chosen from the category of cell aggregates, spheroids, organoids, expiants (islets of Langherans), polymer particles encapsulating cells (organoid covered with a layer of biomaterial), cell-seeded micro-carriers, biomaterial beads.

[0032] According to one variant, the transfer is repeated to manufacture a material, tissue or organ and the volume fraction or volume density of the biological objects transferred by propulsion in said printed material, tissue or organ is greater than 30%.

[0033] According to another variant, the transfer is carried out only once in order to precisely characterize a single object.

[0034] According to another variant, the process is combined with other printing technologies such as extrusion, inkjet, LIFT in order to manufacture complex materials or fabrics comprising different components.

[0035] The invention also relates to equipment for handling and transferring energy by deposition comprising: - a power source directed towards the material to be transferred, - at least one well from which said material is handled and transferred, - a target receiving substrate that collects the transferred material

[0036] Said donor being constituted by a plate having wells containing the transferable objects arranged in a liquid with an orientation Dx,Dy in the plane of the liquid film and Dz perpendicular to said film, characterized in that the transfer takes

[0037] - either by LIFT if the ratio between the dimension (Dz) of the object and the thickness (e) of the film liquid is less than 1

[0038] - either by propulsion if the ratio between the dimension (Dz) of the object and the thickness (e) of the liquid film is greater than 1

[0039] According to one variant, the energy deposition is achieved by focusing a laser for both transfer modes, propulsion and LIFT.

[0040] Advantageously, the substrate of the plate containing the wells is transparent or weakly absorbing at the wavelength of said laser beam.

[0041] According to one variant, it includes a scanner allowing the laser beam to be positioned precisely on the centroid or center of mass of each object, thus ensuring strong directivity in the transfer of said object.

[0042] According to another variant, the energy deposition is achieved via an electric field.

[0043] According to another variant, the energy deposition is achieved by focusing a wave acoustic.

[0044] According to another variant, it includes a control and servo system for the deposited energy value in order to transfer the object optimally according to its size.

[0045] According to another variant, it incorporates means for automating the movement of the substrate (30).

[0046] According to another variant, the substrate of the well plate is covered by a sacrificial layer having strong absorption / conduction properties for energy deposition by laser or by electric field.

[0047] Advantageously, it includes a means for controlling a temporal succession of several energy deposits on the same well for the transfer of an object.

[0048] According to another variant, that it includes a means for controlling several spatially separated energy depots in order to transfer several objects arranged in different wells in parallel.

[0049] According to another variant, it further comprises at least one other printing technology including extrusion, inkjet, LIFT for the manufacture of complex materials or fabrics comprising different components.

[0050] According to another variant, it incorporates a system for delivering several laser beams simultaneously to the object when the latter has a non-isotropic shape in order to guarantee its transfer along a homogeneous trajectory.

[0051] Detailed description of a non-limiting example of embodiment

[0052] The present invention will be better understood upon reading the following description, concerning a non-limiting example of an embodiment illustrated by the accompanying drawings where:

[0053] [Fig. 1] Fig. 1 shows a cross-sectional view of a micro-well of an example of a donor substrate with a small particle

[0054] [Fig.2] Fig.2 shows a cross-sectional view of a micro-well from an example of donor substrate with a large particle

[0055] [Fig.3] Fig.3 represents a schematic view of the transfer system

[0056] [Fig.4] Fig.4 shows a cross-sectional view of a variant of a micro-well

[0057] [Fig.5] Fig.5 represents a partial top view of a donor substrate.

[0058] General context of the invention

[0059] The invention relates particularly to the additive manufacturing of a biological graft by transferring biological objects larger than 100 µm, such as spheroids or organoids, onto a target surface. Spheroids are three-dimensional (3D) cell aggregates that can mimic tissues. Once seeded in a well of a pyramidal or frustoconical microplate, these aggregates form a discrete spheroid.

[0060] The spheroids contain both deeply embedded cells and those with an exposed surface, proliferating and non-proliferating cells, as well as a center surrounded by a well-oxygenated outer layer of cells. Their assembly by transfer onto a target substrate makes it possible to form three-dimensional tissues such as cartilage, for example, to regenerate damaged cartilage.

[0061] Spheroids and organoids can be made up of different stem, progenitor and / or differentiated cells, for example cardiac, cerebral, hepatic cells, etc.

[0062] The culture of spheroids and organoids can be carried out in single-well plates as well as in 12, 24, 48, 96 or even 384-well plates containing several thousand to several tens of thousands of micro-wells.

[0063] General presentation of the donor substrate

[0064] Figures 1 and 2 show views of a micro-well (31) of a donor substrate (30). The micro-well (31) is made in a transparent plate comprising a micro-well matrix, and has an inverted pyramidal shape with an open square base (32) of width L typically between 200 pm and 800 pm and a pointed bottom (33) or preferably with a flat surface. It has a height h typically between 200 pm and 800 pm,

[0065] The spheroid (35) is composed of approximately 500 cells, and occupies only a portion of the height h of the micro-well,

[0066] The spheroid (36) is composed of approximately 2000 cells, and occupies the entire part of the micro-well of height h.

[0067] An aqueous liquid, for example water with added salts or a culture medium, for example a 2% BSA (Bovine Serum Albumin) solution, fills at least partially the micro-well (31).

[0068] Depending on the size of the spheroid (35, 36) that the micro-well (31) contains, the liquid completely covers the spheroid, or the spheroid rests in a liquid background.

[0069] In the first case, the transfer will be carried out in LIFT mode, with a moderate power typically of 15 to 20 microjoules.

[0070] In the second case, the transfer will take place according to a propulsion mode, without the particle being carried away in a liquid bubble, with a power 2 to 5 times greater, on the order of 30 to 60 microjoules.

[0071] The deposited energy value is minimized in order to transfer the object according to its size while ensuring its post-transfer integrity. Minimizing the deposited energy also ensures a low transfer velocity, allowing for slow deposition on the receiving substrate, thus contributing to the integrity of the transferred object.

[0072] Optionally, the surface of the micro-well (31) is coated with a sacrificial layer, typically a layer of gold, to promote the formation of a cavitation bubble ejecting the particle contained in the micro-well. This sacrificial layer can also consist of a thin layer of metal, polymer, gel, etc.

[0073] General presentation of the transfer system

[0074] The transfer system comprises several parts: - an optical section comprising a pulsed laser (10) to generate the cavitation energy of the carrier liquid and optionally a camera (20) for observing the donor substrate (30) and the receiving substrate (40). This camera and the associated optical system are not necessary for firing objects exclusively contained in micro-wells, as the positioning of the objects to be fired is constrained and does not require correction by optical observation. - An automated part, with a robotic arm (50) used for automating the handling of the receiver (40). The receiving substrate (40) advantageously has mechanical properties capable of absorbing shocks in order to guarantee the integrity of the transferred object, either by an elastically deformable coating, or by a support mounted on a damping system along the transfer axis.

[0075] and optionally, the system may include an extruder to add on the receiving substrate (40) a link, for example collagen between the layers of transferred particles.

[0076] More generally, the equipment can combine several 3D printing, bio-printing, and photo-polymerization technologies.

[0077] The substrate (30) consists of a micro-well plate described above, held by a support advantageously moved by a motorized system (37) positioning the tip of one of the micro-wells in the optical axis, to allow the transfer of the particle contained in the micro-well with a high firing accuracy, less than 50 pm, due to the knowledge of the geometry of the plate and the constrained positioning of the particle (35, 36) in the micro-well (31).

[0078] The optical part of the device is possibly made up of two parts, an optional part comprising the camera (20) used to aim at objects, and a part comprising the laser (10) used to "shoot", that is to say to deliver energetic pulses in a plane of the donor micro-well (30) where there is a particle to be transferred.

[0079] The laser (10) is for example an Nd-YAG laser which emits pulses of 1 to 10 ns at 1064 nm with an energy of 15 to 60 microjoules, substantially higher than the energy usually used for LIFT processes.

[0080] According to another example, it consists of a pulsed Ytterbium fiber laser emitting at 1030nm, with shorter pulses, from 350 femtoseconds to 10 picoseconds, with several tens of microjoules of energy per pulse.

[0081] The power will be determined for each shot according to the size of the particle present in the micro-well in the optical axis, and the appropriate transfer mode.

[0082] The lens (16) is typically an F-theta lens with a focal length of 100 mm suitable for laser scanning. The typical size of the spot at the focal plane is on the order of 30 to 35 pm in diameter.

[0083] The laser beam (14) passes through a shaping optic (13) and is then directed via a set of mirrors (11, 12) to a scanner (15) which will then send the beam vertically towards the donor substrate (30), via the F-Theta lens (16).

[0084] The scanner (15) consists of two automated mirrors that redirect the beam horizontally at a certain angle towards the objective lens (16). The objective lens (16) then straightens the beam (14) so ​​that it arrives perpendicular to the donor substrate (30) and focuses it. The mirrors of the scanner (15) control the movement of the laser beam (14) along the horizontal axes on the donor substrate (30). The laser beam (14) is therefore focused on the donor substrate (30) and steerable along the X and Y axes defining the horizontal plane.

[0085] For a donor substrate (30), the beam (14) is focused on the sacrificial layer coating the surface of the substrate, for example a 20 nanometer gold layer deposited on a transparent optical window.

[0086] The second optional optical part comprising the camera (20) is the part used for aiming. It may be necessary to use a visualization system if the particles are smaller than the size of the micro-well used, for example spheroids formed by an aggregation of cells having a diameter of 100 µm.

[0087] A visible light source, typically an LED (21), is placed above the cartridge, and the light beam (22) passes through the scanner (15) following the reverse path of the laser beam. It then reaches a semi-reflective mirror (12) which allows visible light to pass through towards the camera (20), but reflects infrared light back towards the laser (10). The beam then passes through a lens (23), an iris (24) and an objective (25) before arriving at the camera (20). All are aligned so that the focus of the laser beam (10) is at the center of the image captured by the camera (20).

[0088] This image makes it possible to determine the size of the particle present in the micro-well located in the firing axis, and thus determine the power of the pulse required.

[0089] The invention relates, without limitation, to the transfer of spheroids formed by an aggregate of cells cultured in the laboratory and having the appearance of a small pearl made of cells and extracellular matrix.

[0090] In order to produce the spheroids, stem, progenitor or differentiated cells are cultured by conventional methods and seeded in micro-wells in order to produce aggregates which are then manipulated according to the process of the present invention.

[0091] As explained above, the "firing zone imaging" component is optional. It is useful when the micro-wells can accommodate small particles whose positioning will not be constrained by the micro-well walls. In such cases, it is necessary to use the information provided by an imaging system to precisely center, to within 50 µm, the axis of the laser beam with the center of the particle.

[0092] On the other hand, when the particles are systematic and of large size, greater than the mid-height section of the micro-well, this imaging subsystem is optional and can be omitted, because the accuracy of the shot results from the constrained positioning of the particle in the micro-well, and from the precise positioning of the substrate presenting the micro-wells relative to a reference point of the equipment.

[0093] Flat-bottomed micro-wells

[0094] According to an alternative embodiment illustrated in [Fig. 4], the micro-wells have an inverted pyramidal cavity with a truncated tip, to limit the reflection and diffraction phenomena of the laser beam centered on the micro-well. The surface area of ​​the base of the micro-well is greater than the width of the laser beam, typically between 50 and 100 pm.

Claims

Demands

1. - A method for transferring an object from a donor substrate (30) to a target substrate (40), a liquid carrier film containing the particles to be transferred being deposited on said donor substrate (30) containing the objects to be transferred of dimension (Dx, Dy, Dz), said transfer being ensured by local energetic excitation of said liquid to form a cavitation bubble localized at the level of an object, characterized in that a. said donor substrate (30) is constituted by a micro-well plate having a plurality of wells each forming a receptacle having an open upper base narrowing towards the bottom, the width (L) of the base of said wells being L > 2DxDy and the height (h) of said well h > 2DZ, and b.the transfer takes place - According to a first mode of transfer with a first energy level Ei if the ratio between the dimension (Dz) of said object and thickness (e) of said liquid film is less than 1 - According to a second mode of transfer with a second energy level E2> Ei if the ratio between the dimension (Dz) of said object and thickness (e) of said liquid film is greater than 1.

2. - Method according to claim 1 characterized in that the energy deposition is achieved by focusing a laser for a propulsion transfer mode or for a LIFT transfer mode.

3. Method according to claim 1 characterized in that the energy deposition is carried out via an electric field for a propulsion transfer mode.

4. - Method according to claim 1 characterized in that the energy deposition is achieved by focusing an acoustic wave for a propulsion transfer mode.

5. - Method according to claim 1, characterized in that the receiving substrate (40) comprises a damping means along the axis of transfer of objects.

6. - A method according to claim 1, characterized in that the transfer of objects is achieved without contact by the generation of a cavitation bubble in the liquid present between the object and the bottom of the well, the transfer being ensured - either by the conversion of the deposited energy into kinetic energy of the object, which lifts off and leaves the liquid layer with high directionality in the propulsion mode - or by the conversion of the deposited energy into a movement of the liquid, which carries the objects away via a jet in a LIFT transfer mode

7. - Method according to claim 1, characterized in that the objects to be transferred are chosen from the category of cell aggregates, spheroids, organoids, expiants (islets of Langherans), polymer particles encapsulating cells (organoid covered with a layer of biomaterial), cell-seeded micro-carriers, biomaterial beads.

8. - Method according to claim 1 characterized in that the transfer is repeated to manufacture a material, tissue or organ and that the volume fraction or volume density of the biological objects transferred by propulsion in said printed material, tissue or organ is greater than 30%.

9. - Method according to claim 1 characterized in that the transfer is carried out only once for the purpose of precisely characterizing a single object.

10. - A method according to claim 1, characterized in that it can be combined with other printing technologies such as extrusion, inkjet, LIFT in order to manufacture complex materials or fabrics comprising different components.

11. - Energy deposition handling and transfer equipment comprising: - an energy source directed towards the material to be transferred, - at least one well from which said material is handled and transferred, - a target receiving substrate that collects the transferred material The said donor being constituted by a plate having wells containing the transferable objects arranged in a liquid with an orientation Dx,Dy in the plane of the liquid film and Dz perpendicular to said film, characterized in that a. said substrate is constituted by a micro-well plate having a plurality of wells each forming a receptacle having an open upper base narrowing towards the bottom, the width (L) of the base of said wells being L > 2DxDy and the height (h) of said well h > 2DZ, and and in that a.that it includes means of transfer: - either by LIFT if the ratio between the dimension (Dz) of the object and the thickness (e) of the liquid film is less than 1 with a first energy level Ei - or by propulsion if the ratio between the dimension (Dz) of the object and the thickness (e) of the liquid film is greater than 1 with a second energy level E2> Ei And in that it integrates a system to deliver several laser beams simultaneously at the level of the object when the latter has a non-isotropic shape for its transfer along a homogeneous trajectory.

12. - Equipment according to claim 11 characterized in that the energy deposition is achieved by focusing a laser for said two modes of transfer, propulsion and LIFT.

13. - Equipment according to claim 11 characterized in that the substrate of the plate having the wells is transparent or weakly absorbing at the wavelength of said laser beam.

14. - Equipment according to claim 11 characterized in that it comprises a scanner enabling the laser beam to be positioned precisely on the centroid or center of mass of each object, thus ensuring strong directivity in the transfer of said object.

15. - Equipment according to claim 11 characterized in that the energy deposition is achieved via an electric field.

16. - Equipment according to claim 11 characterized in that the energy deposition is achieved by focusing an acoustic wave.

17. - Equipment according to claim 11, characterized in that it comprises a control and servo system for the deposited energy value in order to transfer the object according to its size.

18. - Equipment according to claim 11, characterized in that it incorporates means for automating the movement of the substrate (30).

19. - Equipment according to claim 11, characterized in that the substrate (30) of the well plate is covered by a sacrificial layer having absorption / conduction properties of energy deposition by laser or by electric field.

20. - Equipment according to claim 11, characterized in that it comprises a means for controlling a temporal succession of several energy deposits on the same well for the transfer of an object.

21. - Equipment according to claim 11, characterized in that it comprises a means for controlling several spatially separated energy depots in order to transfer several objects arranged in different wells in parallel.

22. - Equipment according to claim 11, characterized in that it further comprises at least one other printing technology including extrusion, inkjet, LIFT for the manufacture of complex materials or fabrics comprising different components.

Images