Technologies have been developed to pierce or make permeable numerous worms' cuticle in an easy and quick way to transfect DNA into the germ line.
A species of worms commonly used is C.elegans. These worms present a typical size of about 1 mm in length and ⅓ mm in diameter.
One laboratory injection equipment of genetic material commonly used in order to obtain transgenic worms is constituted by a microscope, a micro-injector and a micromanipulator. Microinjection requires long and tedious manual handling and much expertise is essential for the sample preparation, handling and injection equipment. Besides, the injection equipment is expensive.
Another known type of device for microinjection is a grid of micro needles, referred to as a “bed of nails”. This tool allows to pierce/make permeable numerous worms' cuticle in a very easy and quick way in order to apply electroporation and transfect DNA into the germ line. This tool allows wounding a high number of worms in few seconds.
In laboratory, such a device is generally made on a silicon wafer (Si/SiO2) using UV photolithography followed by wet and chemical engraving.
During the wet and chemical engraving, the wafer is often held by a fixed support. Then the assembly of the wafer and the support is introduced in a beaker containing an ad hoc solution of KOH with experimental conditions that allow attack of the surface layer of the wafer according to the laws of crystal physics.
The beaker is placed in a bain-marie on a heating agitator. Commonly used agitator in laboratory scale comprises a coil system that is arranged under the beaker for driving a magnetic agitating member, in particular a magnetic rod, inside the beaker. The KOH solution is thus continuously-mixed by the magnetic rod. However, the reaction produces a release of micro-bubbles which, if not properly evacuated, disturb the etching process. This may lead to an uneven attack of the wafer surface by the KOH solution and may degrade severely the precision of etching.
Numerous other solutions have been proposed to etch wafers, most often at an industrial level for the semi-conductor industry.
JP01201490 discloses rotating back and forth wafers immerged in an etching solution.
CN 2058786U teaches using a wafer holder for rotating a wafer in an opposite direction from an agitating member in an etching liquid.
JP 2001-15482 describes an etching device in which a plurality of vertically disposed wafers are in contact, at the bottom of their peripheral edge, with a magnetic rod. The wafers and the magnetic rod are immersed in an etching solution. A magnetic device situated outside the etching solution drives in rotation the magnetic rod and thus the wafers in a rotational movement in an opposite direction relative to that of the magnetic rod.
Some of these devices are quite complex and expensive and inappropriate at the laboratory scale where only a few wafers need to be etched from time to time.
There thus remains a need for improving devices for etching wafers at the laboratory scale.
In order to further improve the quality of etching, exemplary embodiments of the present invention provide a method of etching a substrate, in particular a wafer, in order to produce a grid of micro-protrusions, the method comprising disposing the substrate on a magnetic supporting device, and driving, in an etching solution, the magnetic supporting device in rotation via a magnetic agitator external to the etching solution so that the magnetic supporting device causes the substrate to rotate at least in a same direction as the magnetic supporting device.
The invention makes it possible to better evacuate, from the surface of the substrate, undesired elements produced by the chemical reaction of etching, such as micro bubbles, and thus improves the quality of the etched surface.
The supporting device and the substrate may be disposed, during the etching, in any container appropriate for containing the etching solution, such as a beaker. The magnetic agitator is preferably disposed under a container containing the etching solution.
The substrate is preferably a planar substrate.
The substrate may be of a largest dimension ranging from 2.5 to 10 cm.
According to one exemplary embodiment of the invention, a plurality of substrates is disposed simultaneously on the supporting device.
The substrate(s) may be disposed on the magnetic supporting device in a static manner, i.e., the substrate(s) may not move relative to the magnetic supporting device before and/or during the etching process.
Alternatively, the substrate(s) may be disposed on the magnetic supporting device in a moveable manner, i.e., the substrate(s) may perform relative movement with regard to the magnetic supporting device while remaining carried by or immobilized thereon. This may allow to set the orientation of the substrate relative to the magnetic supporting device and to help finding the orientation that leads to the best results of the etching process.
The magnetic supporting device may comprise at least one removable part that can be withdrawn from the etching solution without withdrawal of the substrate(s) to be etched. The method of etching a substrate may comprise disposing a test substrate on the one or each removable part. In this way, the progress of the etching can be known by withdrawing and analyzing the test substrate from the etching solution, preferably on a regular basis during the etching process.
The substrate may rotate while fixed on the magnetic support device about an axis perpendicular to the axis around which the magnetic supporting device rotates.
Exemplary embodiments of the invention also provide a system comprising:
a magnetic agitator,
a magnetic supporting device configured for holding at least one substrate to be etched in an etching solution, in particular a wafer, in a predefined configuration relative to the magnetic supporting device, so that when the magnetic supporting device is driven in rotation by the magnetic agitator the substrate rotates in a same direction as the supporting device.
For example, if the magnetic supporting device is driven clockwise in rotation within a beaker or any other container containing the etching solution by the magnetic agitator, the substrate also rotates clockwise relative to the beaker. Similarly, if the magnetic supporting device is driven anti-clockwise relative to the beaker, the substrate rotates anti-clockwise relative to the beaker. The movement of the substrate may comprise further motion components, and may be a complex movement comprising a general rotation together with the support and a further movement.
The invention allows using a magnetic agitator commonly used in laboratory for driving in rotation a magnetic rod placed in a beaker. Thus, the invention makes it possible to achieve good etching results in a simple manner. This is advantageous for making etched wafers at the laboratory scale.
Preferably, the supporting device comprises a discrete magnet, for example a magnetic rod.
In a variant, the supporting device comprises a plurality of discrete magnets.
Exemplary embodiments of the invention further provide a magnetic supporting device comprising:
a body,
at least one holding member for holding at least one substrate in a predefined configuration relative to the body,
at least one magnet fixed relative to the body.
The magnetic supporting device may be configured for holding, individually or simultaneously, substrates of different dimensions.
The magnetic supporting device may comprise a plurality of magnets fixed to the body and/or to the holding member.
The holding member(s) may be fixed or movable relative to the body of the magnetic supporting device.
A movable holding member(s) may allow variation of an orientation of the substrate(s) before the etching process.
The holding member(s) may rotate around at least one axis, for example around an axis perpendicular to the axis around which the body of the magnetic supporting device rotates during the etching process.
The holding member(s) may be configured to grip the substrate(s) in various manners.
The holding member(s) may comprise a base element for supporting the at least one substrate and gripping means configured for holding the at least one substrate onto the base element.
The holding member(s) may comprise gripping means for holding substrates of different dimensions.
A holding member may comprise jaws that contact opposite faces or edges of a substrate.
The holding member(s) may comprise portions projecting above the body. This may allow creating more available space for positioning the substrate(s) in the desired orientation relative to the body, and may allow to increase the number and/or the size of substrate(s) to be etched.
Preferably, the holding member(s) are configured so that the substrate(s) held by the holding member(s) are not in contact with the magnet(s) or the body.
The body may comprise a ring-shaped element at its periphery and a central element formed integrally with the ring-shaped element and extending along a diameter of the body. The central element may comprise holes for accommodating corresponding permanent magnets.
The base element may comprise two arms intersecting at their mid-length and forming a cross concentric with the body, the length of each arm being preferably substantially equal to the outer diameter of the ring-shaped element.
Each half of an arm may comprise a hole for receiving a corresponding permanent magnet. The holes on a same arm are preferably positioned symmetrically with regard to the center of the base element.
The body may comprise a wall portion comprising concentric upper and lower rings.
The base element may be fixed to the upper ring, for example by friction. The base element may be movable relative to the upper ring. In a variant, the base element is molded integrally with the upper ring.
The ring-shaped element may be received in the lower ring, for example in a groove thereof.
In a variant, the wall portion comprises a central bar, extending along a diameter of the lower ring. The central bar may comprise holes, for example in the number of two, for receiving corresponding permanent magnets. In this case, the presence of the ring-shaped element and the central element can be omitted.
The holding member may comprise a wafer support configured for being fixed to the base element, for example by magnetic forces, and to which the wafer is fixed.
The wafer support may have a cross-shaped body comprising two arms of equal length. Magnets may be fixed to a bottom surface of the wafer support. These magnets are preferably situated on the wafer support so as to face corresponding magnets of the base element when the wafer support is superposed to the base element.
In a variant, the wafer support is fixed directly to the upper ring,
The wafer support may be fixed by magnetic force or by friction.
The magnetic supporting device may comprise at least one removable part that can be withdrawn from the etching solution during etching of the substrate(s) and configured for holding a test substrate.
The one or each removable part may comprise a handle and a socket attached at a lower end thereof, the test substrate being disposed on the socket.
The one or each removable part may comprise at least one magnet, for example two, received in the socket. An attractive force between the magnets present respectively on the removable part(s) and the base element allows the removable part(s) to be driven in rotation by the magnetic agitator simultaneously with the body.
The removable part(s) may comprise each a gripping part, at a free end of the handle, for facilitating gripping of the removable part(s) during their withdrawal. The removable part(s) may comprise respective identification elements, preferably of different shape. The identification elements may be arranged above the gripping parts. This allows identification of the test substrate(s) during the etching process.
The body and/or the holding member(s) may comprise a plastic material, for example a polyolefin or polytetrafluoroethylene, or any other material which is inert to the etching solution.
Commonly used etching solutions include, among other, strong acid or basic solutions, for example an aqueous solution of KOH heated at about 75° C.
Specific embodiments of the invention will now be described in some further detail with reference to and as illustrated in the accompanying figures. These embodiments are illustrative only, and not meant to be restrictive of the scope of the invention.
bis shows some details of the holding member of
In the prior art, etching of a silicon wafer W at a laboratory scale was commonly performed by a system as illustrated in
The wafers W are placed, for example vertically as illustrated, in a beaker B comprising an etching solution, for example a heated solution of KOH.
A magnetic rod 2 is disposed in the beaker B, which is placed onto a magnetic agitator A. The agitator A drives the magnetic rod 2 in rotation inside the beaker B, thus stirring the etching solution while the wafers W are being etched.
A magnetic supporting device 1 made in accordance with the invention may comprise as illustrated in
The body 6 may be of a ring shape around an axis X. This axis may be an axis of symmetry for the body 6. The body 6 may comprise respective holes and/or housings for receiving corresponding holding member(s) 3. Preferably, the body 6 comprises holes and/or housings for receiving holding member(s) 3 of different types. This may allow wafers W of different sizes to be held by the supporting device 1.
The magnetic rod 2 may be fixed to the body 6 along a diameter thereof. For example, the body 6 comprises two diametrically opposite holes 21 into which the magnetic rod 2 is inserted, so that rotation of the magnetic rod 2 drives in rotation the body 6 around the axis X.
An external agitator A is used to drive the magnetic supporting device 1 in rotation around the axis X.
The holding member 3 rotates in a same rotational movement as the body 6.
In the embodiments of
The body 6 may comprise as shown two diametrically opposite holes 61 situated close to the upper face thereof for insertion of the rod 33.
At least one end of rod 33 may project radially outside the body 6. This allows a user to turn the rod 33 around its longitudinal axis to set the wafer W with the desired orientation relative to the body 6. In the illustrated embodiment, the two opposite ends of the rod 33 are inserted in respective holes 61. In a variant not illustrated, the rod 33 is held in cantilever fashion with only one end thereof inserted in a corresponding hole 61.
Preferably, the magnetic rod 2 is offset with regard to the holding member 3 along the axis X of the body 6. The magnetic rod 2 is situated beneath the holding member 3 so that it does not come into contact with the wafer W to be etched.
The wafer W may have in front view a polygonal, for example square shape, as illustrated in
In a variant, the wafer W may have other shapes, such as for example a circular outline as illustrated in
The rod 33 may comprise a planar top surface 34 to provide support for the rear face of the wafer W, as illustrated in
The rod 33 may be tightened onto the body 6 at the desired orientation by at least one screw 35 introduced in a corresponding fixing hole 38.
In the embodiment of
In a variant, the base element 31 projects above the body 6. Having the base element 31 projecting above the body 6 may allow creating more available space for positioning the wafer W, thus increasing the number and/or the size of wafers to be etched. This may also allow the wafers to be etched simultaneously on both faces.
In the variants illustrated in
This shape of the base element 31 may provide more supporting surface in contact with the wafer and may improve the stability of the wafer W relative to the base element 31 during the etching.
The base element 31 may comprise stops 42 for holding the wafer W at its periphery.
Preferably, the size of the base element 31 is chosen to match that of the wafer W to be etched.
The frame 37 may not extend beyond the periphery of the body 6 as illustrated in
The diameter of the body 6 may be chosen as a function of the size of the wafer W to be etched and may range between 75 and 100 mm, for example.
The thickness e of the body may lie between 0.7 mm and 10 mm, for example. The height h of the body 6 may range from 15 mm to 30 mm.
During the etching process, the main faces of the wafers W may be oriented perpendicular or oblique to the axis X of the body 6.
The holding member 3 may comprise a rotational support 39 allowing setting the orientation of the wafer W about an axis Y perpendicular to the axis X of the body 6. The axis Y is for example parallel to a diameter of the body 6.
The rotational support 39 may be configured so that an angle a between the upper face of the wafer W and the axis X of the body 6 lies between 0° and 20°, as shown in
The base element 31 may be frictionally held by the rotational support 39. The gripping element 32 may comprise next to the rotational support 39 a jaw made by a slot 36 into which the wafer W is introduced and a clamping screw 43 for immobilizing the wafer W in the slot 36, as shown in
The rotational support 39 may comprise a cylindrical sleeve 41 defining a bearing for a tip 47 of the gripping member 32. The sleeve 41 is connected to the body 6 via a stem 45 fitted into a corresponding housing 46 opening out on the upper face of the body 6. The body 6 may comprise four housings 46, disposed equally around the axis X of the body 6. For example, as shown, the diametrically opposite holes 21 for insertion of the magnetic rod 2, the diametrically opposite holes for insertion of the rod 33 and the housings 46 may be equally spaced around the axis X of the body.
In the embodiment of
In the variant of
The cooperation between the gripping members 32 and the rotational supports 39 allows rotation of the wafer W relative to the body 6 around an axis Y perpendicular to the axis X of the body 6 in the predefined range.
In a variant not illustrated, the holding member 3 does not comprise a frame for supporting the wafers. For example, in case of wafers of small size, the wafers may be held in the slot 36 of the gripping member 32. The holding member 3 may not comprise a frame.
In the variant illustrated in
The body 6 may receive thanks to the housings 46 up to four such holding members.
Preferably, the wafers W are disposed on the body 6 in a manner that the wafers are not in contact with each other.
Preferably, the wafers W do not overlap, when viewed from above along the axis X, as illustrated in
The gripping elements 32 may be configured for disposing the wafers W at a same height along the axis X of the body 6.
In a variant, the gripping elements 32 are configured for holding the wafers W at different heights along the axis X.
In the embodiment shown in
The body 6 may be closed at its lower end by a bottom wall 13, as illustrated in
To use a magnetic supporting device made in accordance with the invention, one attaches the wafer W with the gripping means and places the assembly in a beaker B containing the etching solution, for example a solution of KOH as shown in
In the variant shown in
In this embodiment, the base element 31 comprises two arms 30 intersecting at their mid-length and forming a cross concentric with the body 6. The length L of each arm 30 is substantially equal to the outer diameter of the ring-shaped element 66. In the illustrated embodiment, each arm 30 has, when viewed along a longitudinal direction thereof, a rectangular cross section with a large side parallel to the plane along which the ring-shaped element 66 extends.
As illustrated in
The magnets 20 received by the central element 65 are mainly responsible for the rotational movement of the magnetic support device. The magnets 20 preferably have a diameter ranging from 4 to 8 mm, for example around 6 mm, and are capable of holding a weight of around 1 kg, for example between 1 and 1.5 kg.
The magnets 60 received in the base element 31 are mainly responsible for the attraction of the base element 31 with other elements of the holding member, for example a wafer support 10 or a removable part 70, as explained further with respect to
The magnets 20, 60 may have a circular cross section. In a variant, the magnets have a substantially parallelepipedal shape, as illustrated in
The base element 31 is fixed to the ring-shape element 66, for example by screwing, at end portions 77 thereof. The end portions 77 comprise protrusions 93 that abut against the upper surface of the ring-shaped element 66.
As illustrated in
In the embodiments of
In the variants illustrated in
The base element 31 may comprise holes 62 for immobilizing the gripping elements 32 or pins 44 at predefined positions, as visible in
The gripping elements 32 may be fixed to the base element 31 by snap-fastening. Each gripping element 32 may have a general U shape as shown in
More specifically, in the embodiment of
In the variant illustrated in
In the embodiment of
As shown in
In the embodiment of
In the variant illustrated in
The support 10 comprises, at three respective end portions 78 of the arms 17, gripping protrusions 88. Each protrusion 88 extends along an angle γ with the wafer support 10 and defines a notch against the bottom of which the wafer can abut.
The remaining half of the arm not provided with a corresponding protrusion is configured to allow a gripping insert 90 to be fixed thereon. In the shown example, this arm half comprises a hole 72 for fixing the gripping insert 90.
As can be seen in
In use, the wafer W is first positioned against the gripping protrusions 88. The gripping insert 90 is then introduced into the hole 72 and the flange 92 covers the peripheral edge of the wafer W. The member 90 can be turned so that the shoulder 94 abuts against the edge of the wafer and immobilizes it.
As shown in
The magnetic supporting device may comprise at least a removable part 70 configured for holding a test substrate T during the etching process, as shown in
In the embodiment illustrated in
In use, a test substrate T is fixed to the removable part 70 via gripping means 32. The removable part 70 is introduced in the etching solution in which the base element 31 holding the substrate W to be etched is placed and attracted thereto by magnetic force. When the magnetic agitator is turned on, the test substrate T rotates at the same speed as the substrate W held on the base element 31.
In order to check the progress of the etching, the user may turn off the agitator, for example by a few seconds, and withdraw the test substrate T from the etching solution via the handle 71. The use of a magnetic force to fix the removable part 70 to the base element 31 allows removal of the removable part 70 to be performed in a rapid and smooth way without friction between different parts of the device. The removable part 70 can be detached from the base element 31 by a simple rotational movement around an axis passing by the intersection of the socket 72 and the handle 71 and perpendicular to a main plan N of the removable part. The attractive force between the removable part 70 and the base element 31 is chosen to ensure that withdrawal of the removable part 70 does not result in withdrawal of the base element 31 and body 6, which remain in the etching solution. The magnet 60 received in the removable part 70 may have a same or similar dimension and force of traction as those received in the base element 31. For example, the magnets 60 are capable of holding a weight of around 0.15 kg.
After analysis of the test substrate T, the latter may be reintroduced into the etching solution via the removable part 70 if necessary. Withdrawal of the removable part 70 can be performed on a regular basis during the etching process.
In the embodiment illustrated in
In the device illustrated in
The socket 72 may comprise as shown in
During the etching, the removable parts 70 can be removed one by one, for example on a regular basis.
In the variants illustrated in
The removable part 70 may comprise an identification element 99, as illustrated in
The identification elements 99 may be of any shape that allows the user to differentiate the different removable parts 70. For example, the identification elements 9 may be in the form of letters or numbers.
In the variant illustrated in
In the embodiment illustrated, two opposite connecting elements 64 comprise each a hole 61 configured for insertion of a base element 31 in the form of a rod 33, as the one illustrated in the examples of
The upper ring 67 comprises holes 48 for receiving the protrusions 93 of the base element 31. By properly choosing the height of the protrusions 93 and/or the depth of the holes 48, the base element 31 may form an angle β with the wall portion 63, preferably ranging between 2° et 4°. In a variant not illustrated, the base element 31 is fixed to the wall portion 63 by magnetic force. In another variant, the base element 31 is movable relative to the upper ring 67 so as to allow adjustment of the angle β.
The lower ring 69 comprises a groove 96 in which the ring-shaped element 66 is received. Two opposite connecting elements 64 are situated along the central rod 65.
In a variant, the wall portion 63 comprises a central bar 56 extending along a diameter of the lower ring 69, as in the embodiment of
The substrate W is held to the base element by magnetic force via a wafer support 10 as described with regard to
In the variant illustrated in
In a variant not illustrated, the base element 31 is integrally formed with the wall portion 63.
A 2 L beaker containing 500 mL of water is heated at 75° C. 150 ml of 40% KOH solution is added to a 400 mL beaker, which is transferred to the 2 L beaker in order to be heated in water bath. Heating is kept until the temperature of the water in the 2 L beaker reaches 75° C. again.
The magnetic supporting device of
In each case, a wafer comprising a homogeneous set of microstructures M is obtained.
Two substrates each of 1 cm2 having followed UV photolithography are used. The UV photolithography was performed using respective masks with hexagonal networks as illustrated in
As illustrated in
Etching is performed with an etching solution of KOH having a concentration of 45% at 70° C., with the magnetic support device turning at a speed of 190 rpm.
The two test substrates T are respectively etched during 50 and 60 minutes. The results of etching are illustrated on
Both test substrates show good homogeneity.
The invention is not limited to the embodiments described above.
For example, the magnetic rod may be fixed to a bottom wall of the device.
Holding members other than those described above may be used for holding one or more wafers with or without allowing relative movement between the wafers and the body of the device.
The device according to the invention may be used for etching silicon wafers for various applications, not only for preparing microneedles to treat worms. The device may also be used for etching substrate other than silicon substrates.
For example, the invention may be used in all industries wherein wet and chemical engraving of wafers, in particular by KOH, is performed following UV photolithography for creating microstructures thereon. Such industries may include among other electronics, for example for the production of printed circuit, or pharmaceutical industries wherein wafers with micro-needles thereon may be used.
Number | Date | Country | Kind |
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17305019.6 | Jan 2017 | EP | regional |
Filing Document | Filing Date | Country | Kind |
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PCT/EP2018/050270 | 1/5/2018 | WO | 00 |