Integrated glass reactor, manufacturing method, and analytical method

Abstract

To provide a single-piece reaction vessel made of glass, a production method, and an analysis method.SOLUTION: The invention relates to a method of production of glass reaction vessels, and to a glass reaction vessel obtained by the method. The method includes: 1. irradiating a surface of a first glass plate with a laser beam of a wavelength to which a first glass plate is transparent; and 2. etching the first glass plate; 3. where the etching of the first glass plate is terminated when recesses extend, over only a portion of the thickness of the first glass plate and therefore the recesses have a bottom formed in the first glass plate as a single piece.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a method for manufacturing a reactor made of glass, which is formed in a concave shape in the glass. Since the glass in which the reactors are formed in an array has an integral structure, the reactors are formed in an integral glass, particularly a glass plate. The method forms an array of a large number of reactors, and the array is formed in the glass having an integral structure. For example, an array of 25×25 meshes of reactors of 8×12 each is formed. A large number of the reactors are formed in a glass plate made of an integral structure of a first glass plate.

Background Art

[0002] Since the method forms the recesses for forming the reactors without applying a mechanical action to the massive glass, the reactors have an advantage that they do not have mechanical damages such as micro cracks. The reactors have a large aspect ratio of the depth to the diameter. A further advantage is that the method for manufacturing the reactors can proceed at least without performing a selective coating of the glass surface where the cross-sectional opening of the reactor is formed, and optionally without performing any coating of the glass surface where the reactor is formed.

[0003] Non-Patent Document 1 describes the manufacture of a reactor having a diameter of 20 μm and a depth of 8 μm in a glass plate by etching after applying a mask made of chromium and a photoresist thereon.

[0004] Patent Document 1 describes the manufacture of a reactor in glass using a tool irradiated with ultrasonic waves and an abrasive between the tool and the glass.

[0005] Patent Document 2 describes a microtiter plate having 96 wells made of the bottom of glass, the bottom transmitting ultraviolet rays and being connected to a glass plate by frit glass, and in the glass plate, continuous recesses form the side walls of the wells.

[0006] Patent Document 3 describes the formation of a surface structure at the bottom of a microtiter plate using a photolithography method.

[0007] Patent Document 4 describes a method for etching glass by irradiating a glass body with a laser at a wavelength absorbed by the glass for 1-2 seconds (paragraph

[0037] , e.g., line 3) such that the glass is heated above its strain temperature and swells at the irradiated line (paragraph

[0033] ). Subsequent etching of the glass at the irradiated location is performed using 50% HF / 5 min (paragraph

[0057] , line 11). A reference to the prior art (paragraph

[0005] , last sentence) suggests that an alternative approach is to use a high repetition femtosecond laser with water or another liquid in contact with the glass to clean the removed glass as the damaged area is formed.

[0008] Patent Document 5 describes a method of removing material from the irradiated glass portion by irradiating glass, preferably borosilicate glass, and then etching it using an etching solution. In etching, the glass is immersed in an acid bath agitated with ultrasound, and the etching solution is applied to the glass by rotating the glass in the bath. Optionally, the glass surface can also be polished to remove surface roughness (Column 2, lines 26-49). The etching solution used in the etching bath is hydrofluoric acid (Column 6, lines 31-36), and the flow of hydrofluoric acid on the glass substrate eliminates the accumulation of insoluble by-products of the chemical etching reaction on the surface.

[0009] Patent Document 6 (Nippon Sheet Glass Co., Ltd.) describes a method of processing glass by irradiating it with a laser pulse that absorbs the irradiated wavelength (paragraph

[0008] ), and then etching the glass (paragraph

[0009] , lines 5-6). For etching, HF, sulfuric acid, nitric acid, hydrochloric acid, or a mixture thereof can be used (paragraph

[0037] ). In addition, a film to prevent etching may be formed on the upper or back side of the glass plate so that etching is performed on only one side of the glass plate (paragraph

[0039] , lines 10-14). When etching only the back side, Silitect-II is applied to the upper side of the glass plate for protection (paragraph

[0059] ), then immersed in hydrofluoric acid (paragraph

[0060] ), and then Silitect-II is removed (paragraph

[0061] ).

[0010] Patent document 7 describes a method for generating microstructures in glass by ablation of the glass portion using laser irradiation. It does not mention the need to clean the glass after generating the microstructures. [Prior art documents] [Patent Documents]

[0011] [Patent Document 1] U.S. Patent Application Publication No. 2003 / 0211014 [Patent Document 2] European Patent Application Publication No. 1867612 [Patent Document 3] European Patent Application Publication No. 2011857 [Patent Document 4] U.S. Patent Application Publication No. 2010 / 0050692 [Patent Document 5] U.S. Patent No. 5,919,607 [Patent Document 6] U.S. Patent Application Publication No. 2009 / 0013724 [Patent Document 7] U.S. Patent Application Publication No. 2009 / 0261082 [Non-patent literature]

[0012] [Non-Patent Document 1] Deutsch et al., Lab Chip, 2006, 69, 995-1000 [Overview of the project] [Problems that the invention aims to solve]

[0013] The object of the present invention is to provide an alternative manufacturing method suitable for manufacturing reactors, particularly those with large aspect ratios, in glass in generally small volumes and for supplying arrays of such reactors within the glass. Preferably, the method should be suitable for supplying arrays of such reactors that, when the glass is irradiated with light, they form high optical contrast with the cells within the reactors. [Means for solving the problem]

[0014] The present invention solves the present problem with the features described in the claims, in particular with a method for manufacturing a glass reactor and a glass reactor obtained by said method. 1. A step of irradiating the surface of a glass plate with a laser beam of a wavelength that penetrates the glass plate, preferably in a point-like manner, and more preferably perpendicularly, to locations on the surface of the glass plate where recesses are to be formed as reactors, wherein the laser beam preferably consists of laser pulses having a pulse length of, for example, up to 100 μs, and the focal positions of the laser pulses are set such that the focal points do not extend across the entire thickness of the glass plate, particularly in the direction of laser beam propagation, and the laser pulses are irradiated at multiple spaced positions on the first glass plate where recesses are to be formed. 2. A step of etching a glass plate to form a recess, preferably having a depth of at least 40 μm or at least 30 μm, and preferably having an aspect ratio of at least 2, at least 4, at least 5, or at least 6 in depth to a diameter measured on the plane of the first surface, etching the glass plate over a sufficient time length to form a recess along the location. It has a recess that extends only over the thickness of the first glass plate. Therefore, when the recess has a bottom integrally formed with the first glass plate, the etching of the glass plate ends. Preferably, the bottom of the recess forming the reactor has a recess at any position irradiated with a laser pulse, or is formed by a recess at any position irradiated with a laser pulse, or is constituted by a recess.

[0015] Setting the focal position of the laser pulse so that the focus does not extend over the entire thickness of the glass plate, particularly in the propagation direction of the laser beam, can be obtained by setting the focal position of the laser pulse so that the focus extends only over the thickness of the glass plate, for example, over the first thickness portion of the glass plate, particularly in the propagation direction of the laser beam.

[0016] At this time, any glass plate may be an object made of glass having a lateral spread larger than the thickness. That is, the glass plate may have an outer periphery of a rectangular, circular or other shape that limits the opposite surface.

[0017] The reactor is formed by a recess in the glass plate, and the recess has exactly one opening located in the plane of the first surface of the glass plate. The use of the reactor is not limited to chemical reactions and also includes biochemical, biological and physical processes. This may be a process using each cell, bacterium, virus, protein, etc., which can be an animal cell, a plant cell or a yeast cell, or a cluster of these.

[0018] Dot-like irradiation is obtained by focusing the laser beam onto a point having a size of several micrometers, for example from 1 μm to 10 μm, or up to 5 μm. At this time, it is advantageous if the focus of the laser beam extends over a length much larger than the Rayleigh length of the corresponding laser beam having a Gaussian distribution along the propagation direction of the laser beam. This can be obtained by a suitable optical device such as a diffractive optical element, for example. The irradiation may be an irradiation that does not pass through the glass plate and does not penetrate very deeply up to the first thickness portion in contact with the first surface of the first glass plate. For example, the focus of the laser beam does not extend over the entire thickness of the glass plate in the propagation direction of the laser beam, and the laser beam only extends from the first surface of the glass plate into the first thickness portion in contact with the first surface and terminates within the first thickness portion. Therefore, the laser beam does not extend into the second thickness portion following the first thickness portion of the glass plate. Since the interaction between the laser beam and the material of the glass plate occurs only at the focus, it is possible to terminate the interaction region inside the first thickness portion of the glass plate. Preferably, the laser beam is composed of laser pulses.

[0019] When there is no coating made of an etching resist between the first surface of the first glass plate and the second surface on the opposite side of the first surface, during etching, the regions where the laser irradiates the glass plate and the regions where the laser beam is farther away on the opposite side corrode significantly faster than the adjacent regions. Therefore, the region of the first surface of the glass plate corrodes relatively slowly and in the same shape at intervals from the location of the dot-like laser irradiation because, for example, there is no coating made of an etching resist. Therefore, the first surface is formed by a surface portion arranged in a plane that is the starting point when the recesses extend into the glass volume of the glass plate, excluding the recesses. The surface portion arranged in a common plane that forms the first surface, excluding the recesses, is formed by the end faces of the walls located between the recesses.

[0020] In general, optionally, in a method having or consisting of such a step, an etching resist may be coated over the entire second surface of the glass plate, thereby preventing the formation of recesses from the second surface into the glass plate or uniform corrosion from the surface. According to the present invention, the pulsed laser beam is controlled, for example, by setting the distance of the focal position to the first surface of the glass plate, such that, upon irradiation, the intensity is sufficient only to correct the glass plate along the optical path so that it traverses a first portion of the thickness of the first glass plate. For this purpose, for example, the focal position of the laser pulse is set so that the laser pulse enters the first glass plate only within the thickness portion.

[0021] Optionally, the laser beam irradiated in a point manner in step 1 is positioned on the plane of the first surface of the glass plate, at a distance of at least or exactly the diameter of one of the reactors, including the thickness of the wall between the reactors, to etch recesses where reactors are to be formed in step 2. The diameter of the reactors can be set by the reaction conditions and the duration of etching, because the etching is performed concentrically around the straight path the irradiated laser beam takes through the glass plate. The wall positioned between the reactors terminates in a common plane. The end faces of these walls are located in the common plane and form a first surface, or form a first surface in the common plane, where the first surface is interrupted by recesses. Preferably, the first surface is interrupted only by the cross-section of the recesses.

[0022] The laser beam preferably has a wavelength of, for example, 1064 nm to 515 nm at any point on the glass plate where the laser beam is irradiated, and is pulsed, preferably with a pulse length of up to 100 ps or up to 50 ps, ​​preferably up to 10 ps. In general, the laser is adjusted so that the laser beam does not collide with the glass plate between those points. Preferably, the laser beam is irradiated in a point-like and perpendicular manner on the surface of the glass plate. Preferably, the surface irradiated by the first laser beam forms the first surface of the first glass plate.

[0023] Generally speaking, glass plates are also called first glass plates.

[0024] Etching is carried out using, for example, 1% to 48% by weight of hydrofluoric acid, and / or sulfuric acid, and / or hydrochloric acid, and / or phosphoric acid, and / or nitric acid, or potassium hydroxide, at a temperature of, for example, up to 140°C.

[0025] The glass plate may have a thickness of up to 1000 μm, preferably from 100 μm to 1000 μm, for example up to 800 μm, or for example from 300 μm to 500 μm, before etching, and after etching, it may have a thickness that is 50 μm to 700 μm smaller, for example down to 200 μm.

[0026] In step 3, the etching of the glass plate is completed when the recess extends only to the thickness of the glass plate, the depth of the recess in the first glass plate is only the thickness of the glass plate, and therefore the recess has a bottom integrally formed within the glass plate.

[0027] The recess preferably extends in a conical or frustoconical shape at an angle of, for example, 0° to 15°, starting from the first surface of the glass plate and extending within its volume.

[0028] Optionally, the glass plate can be etched without coating, for example, without using a mask and / or etching resist. Therefore, this method has the advantage of being carried out without applying an etching resist to the glass plate and without removing the etching resist from the glass plate. Generally, at least the first surface of the glass plate is etched without using an etching resist, without having an etching resist or a mask.

[0029] In general, optionally, and especially in the method having steps 1 to 3, or the method consisting of steps 1 to 3, the second surface of the glass plate is completely coated with etching resist so that etching is performed only from the first surface, or the laser irradiating the laser beam is set so that the laser beam traverses only the thickness of the glass plate, or the laser beam terminates within the thickness of the glass plate. In this embodiment, the laser beam is irradiated in a dot pattern at multiple spaced positions, for example, at least two, at least three, at least 10, or at least 30 positions, where recesses are to be formed in the glass plate, and the laser beam is irradiated continuously or simultaneously, preferably parallel to each other and perpendicular to the glass plate. These positions form locations where etching corrodes the glass plate faster than in surface areas away from the position. The laser-irradiated locations result in a constant rate of corrosion of the glass during etching, together forming recesses. The positions irradiated in the area of ​​location and forming locations are arranged, for example, at intervals of 1 μm to 10 μm. Preferably, the locations are situated within the region around each location where a recess is to be formed. Preferably, the locations around the locations where the laser beam is irradiated, or the locations irradiated to form the locations, are spaced apart by 1 μm to 10 μm, for example, 2 μm to 5 μm, or 3 μm, and the spacing is determined in particular within the plane of the first surface of the first glass plate.

[0030] In general, recesses can be formed by individual laser pulses or multiple laser pulses. In the case of individual laser pulses, the diameter of the recess is determined in particular by the duration of etching. In the case of forming recesses using multiple laser pulses, the diameter of the recess is determined by the number and spacing of the positions where the laser beam is irradiated for one location and penetrates the first glass plate. The depth of the recess within the volume of the first glass plate can be determined by the duration of etching, as well as by the fact that the laser beam penetrates the glass plate only by its thickness and does not pass through the glass plate completely.

[0031] Optionally, a laser beam formed by, for example, continuously irradiated laser beam pulses is irradiated around each location where a recess is to be formed, preferably in an annular, rectangular, or hexagonal circumferentially closed path. In this case, the laser beam pulses may be irradiated along the circumferentially closed path to the first glass surface at intervals of 3 μm between laser beam pulses, for example, determined on the first surface of the first glass plate. Thus, optionally, the glass plate can be irradiated with laser beams in a point-like manner at multiple spaced positions at each location, and a laser beam formed by continuously irradiated laser beam pulses around these locations can be irradiated along the circumferentially closed path. Irradiation with a laser beam along the circumferentially closed path has the advantage that, during subsequent etching, a recess is formed having a wall extending from the first surface and having a cross-section surrounding the circumferentially closed path.

[0032] Therefore, the glass plate may integrally have a reactor as a recess. In this embodiment, the bottom of the reactor is formed from the material of the glass plate. Preferably, the bottom of the reactor is formed by a number of adjacent recesses, which are arranged adjacently in a plane that is parallel to, for example, the first surface of the glass plate and the plane of the second surface.

[0033] In this embodiment, the bottom of the recess may have a microstructure configured to provide near-field illumination to the volume of the recess. Such a microstructure may have, for example, the shape of a thin, tall glass tip and may function as an optical waveguide for illumination or may influence individual cells or clusters of cells in their location or position. Such a structure may be formed, for example, by etching a number of locations that are densely irradiated with laser pulses. If each laser pulse is emitted into the center of the recess, the glass tip remains in the recess after the etching process. In general, in this method, recesses can be formed by irradiating adjacent locations with laser pulses followed by etching, where the locations are spaced at the same maximum distance of 10 μm from each other, for example, from 1 μm to 5 μm or from 3 μm, and together form locations, with at least two or at least three locations spaced at larger intervals, for example, from 10 μm to 30 μm or from 15 μm to 20 μm. At least two or three locations of the laser pulses, which are spaced relatively far apart, have, for example, regions between them where the laser pulses are emitted at the same intervals, or surround regions where the glass tip remains after etching.

[0034] In general, laser pulses can be irradiated into the glass plate at various depths at locations that form recesses to be formed by etching. For example, a laser pulse can be irradiated more deeply into the thickness of the glass plate at a given location, allowing for deeper penetration, while other laser pulses can be irradiated to a smaller depth within the thickness of the glass plate at a given location. During subsequent etching, deeper or wider recesses are formed at locations where the laser pulse was irradiated more deeply into the glass plate, and the bottoms of shallower recesses are formed at locations where the laser pulse was irradiated to a smaller depth into the glass plate. In general, depending on the spacing of the locations, a concave depression can be formed at the bottom at each location. The bottom and the recess with further deeper recesses within the bottom can be formed by irradiating the glass plate to a smaller depth with the laser pulse at the location where the bottom should be formed, and by irradiating the glass plate to a deeper depth with the laser pulse at the location where further recesses extending deeper into the glass plate from the bottom should be formed, and by subsequent etching. Due to the larger diameter of further recesses extending deeper into the glass plate, starting from the bottom of the recess, the laser pulses irradiated deeper into the glass plate may be spaced at intervals of, for example, 1 μm to 10 μm, 2 μm to 5 μm, or 3 μm in adjacent positions, thereby allowing etching to penetrate deeper into the glass plate at these positions. If the laser pulses can irradiate the glass plate to a smaller depth in some positions and deeper in others, during etching, the bottom of the concave recess will be formed to a smaller depth at the positions where the laser pulses irradiated to a smaller depth, and further recesses extending deeper into the glass plate will be formed at the positions where the laser pulses irradiated to a deeper depth.

[0035] In the embodiment for forming a reactor formed on a single glass plate or composed solely of glass plates, etching is carried out for a sufficient duration to obtain the desired depth of the recess within the glass volume of the glass plate, which is spaced apart from the second surface, or for a duration only such that the glass plate can still have a closed second surface. Optionally, the bottom of the reactor may have concave depressions having a parabolic or conical cross-section. Such concave depressions can be formed at each location where the laser beam is irradiated point by point. Preferably, such concave depressions have a cross-sectional opening and depth of several micrometers, for example, 1 μm to 5 μm. Preferably, each recess or the bottom of each reactor has at least three concave depressions.

[0036] In embodiments where the entire surface of the second surface of the first glass plate is coated with an etching resist, etching acts only from the first surface. The coating on the second surface results in the formation of a recess that forms a cylindrical or conical shape in the direction from the first surface toward the second surface, preventing etching corrosion of the second surface, so that the recess extends only as far as the thickness of the glass plate. In these embodiments, etching can generally act only on the first surface of the glass plate until the recess extends within the plane of the second surface of the glass plate. When the second surface is coated with an etching resist, it is clear that etching forms a recess as far as the thickness of the glass plate, and the walls of the recess are perpendicular to the second surface or at a conical or frustoconical angle.

[0037] In general, the recesses extend within the glass plate perpendicular to the first surface, and the glass plate, with respect to each recess, forms a thickness portion as an integral bottom structure with respect to the first surface.

[0038] The reactor has a depth of, for example, at least 40 μm, at least 50 μm, or at least 100 μm or at least 150 μm, for example up to 250 μm, or up to 200 μm. The reactor has a diameter of, for example, at least 10 μm or at least 30 μm, for example up to 200 μm, or up to 1 mm, and generally preferably has an aspect ratio of depth to diameter of at least 2, at least 4, at least 5, or at least 6. The recess in the first glass plate has a volume of, for example, 1 pL to 1 μL inside the first glass plate.

[0039] The recesses formed in the glass plate may have a larger cross-section in the upper thickness portion 20 adjacent to the first surface than in the lower thickness portion 21 adjacent to the upper thickness portion 20 and divided into at least two partial recesses. Optionally, partial walls 22 integrally formed from the glass plate to the lower thickness portion 21 are positioned between the partial recesses extending into the lower thickness portion 21. These partial walls 22 are spaced apart from each other around the partial recesses. These partial walls 22 are formed by irradiation with laser pulses that penetrate at most only into the upper thickness portion 20 of the glass plate in the region of the partial walls 22, and by irradiation with laser pulses that penetrate deeper into the region of the partial recesses, and subsequently by etching of the glass plate. During etching, a recess 2 is formed in the upper thickness portion 20 adjacent to the first surface 4 of the glass plate, and the recess 2 extends over the region of at least two partial recesses 2'. The longitudinal central axes 7 of the partial recesses 2' may be spaced, for example, at intervals of 10 μm to 100 μm. The partial walls 22 extend across the lower thickness portion between the partial recesses 2', and the partial recesses 2' may have a diameter of, for example, 1 to 50 on the plane of the second surface of the glass plate 1. Here, the upper thickness portion 20 is also referred to as the first thickness portion, and the lower thickness portion 21 is also referred to as the second thickness portion. The upper thickness portion 20 and the lower thickness portion 21 extend independently of each other, starting from the first surface 4 of the glass plate 1, to, for example, at least 20%, at least 30%, at least 40%, or at least 50% of the original thickness of the glass plate 1, up to 80%, 70%, or 60%. In this case, the lower thickness portion 21 extends within the thickness of the glass plate 1 to a degree that is smaller or larger than that of the upper thickness portion 20. In general, the thickness portions 20 and 21 do not extend across the entire thickness of the glass plate 1. For example, the first thickness portion 20 and especially the second thickness portion 21 may be predetermined to remain present in the recessed area after etching for at least 5%, at least 10%, at least 15%, or at least 20% of the thickness of the glass plate 1, or between the recess 2 and the second surface 5.

[0040] Furthermore, the present invention relates to an analytical method in which light is irradiated onto a reactor inside a glass container and light originating from the reactor is detected, and to the use of a glass reactor in said analytical method.

[0041] In this case, the light for analysis may preferably be shone approximately perpendicular to the glass plate onto the first surface of the glass plate and / or the open recess of the glass plate, or the second surface of the glass plate opposite to the first surface.

[0042] It has been shown that recesses, particularly those extending conically from the first surface into the volume of the glass plate, form a clear contrast with their walls and the bottom of the recess when illuminated with visible light. Therefore, during light detection, the outer periphery of the recess is recorded and can be depicted as a dark ring in contrast to the bottom of the recess.

[0043] The present invention further relates to an analytical method comprising the steps of supplying a reactor manufactured according to the manufacturing method according to the present invention, or a reactor according to the present invention; placing a sample, such as a cell-free sample like plasma, or a cellular sample like complete blood or cells separated from complete blood, or a patient sample which may be tissue material, in advance, simultaneously, or later; adding at least one reagent to the reactor; and performing an analysis. The reactor manufactured by this method and the reactor according to the present invention have the advantage that the bottom of the reactor has a concave depression, and cells can be individually placed in the concave depression.

[0044] Optionally, at least one reagent may be added to multiple reactors in different amounts, either once or multiple times in succession. The reagent may be, for example, a drug-active substance, and the analysis may include measuring the effect of the drug-active substance on a sample. Optionally, the method may include one or more culture steps, for example, under cell culture conditions (37°C, 5% CO2 atmosphere, static or dynamic).

[0045] The analysis may be, for example, optical measurement of the reactor during or after sequencing of DNA and / or RNA and / or proteins in the reactor after adding reagents for sequencing and optionally after cell lysis, or determination of proteins after reaction with binding molecules such as antibodies, preferably marked with a dye, added as reagents. Preferably, the analysis includes, for example, determination of transcribed RNA and / or translated proteins in a sample without added active ingredients, compared with a sample to which the active ingredient has been added. Optionally, the analytical method may include removal of a portion of the sample from the reactor, and further optionally, injection of the removed portion of the sample into a further reactor according to the present invention or a reactor manufactured in accordance with the present invention. The addition of the sample and / or reagent to the reactor may be, for example, by the motion of droplets of the sample and / or the motion of droplets of the reagent, which are formed and moved, for example, as part of a fluid jet. This is carried out, for example, by electromagnetic radiation, irradiation with sound waves or ultrasound, application of an electric field, or pressurization. Droplet formation is known as inkjet printing or pipetting. Alternatively, laser-based printing methods such as laser transfer printing can be used. Preferably, the addition of the sample and / or reagent is carried out without contact between the weighing device and the reactor.

[0046] In embodiments where the conductor path is located in a recess, the analysis method may involve applying a voltage to the volume of the recess and measuring the electrical parameters present within the volume between the conductor paths. [Brief explanation of the drawing]

[0047] [Figure 1] This figure schematically shows an embodiment of a reactor in a one-piece glass plate structure, with a cross-section perpendicular to the surface of the first glass plate. [Figure 2] This figure shows a schematic top view of the first surface of a glass plate, illustrating the optical analysis of a reactor manufactured according to the present invention. [Figure 3a]This figure schematically shows a further embodiment in a cross-section perpendicular to the surface of the first glass plate. [Figure 3b] This figure schematically shows a further embodiment in a cross-section perpendicular to the surface of the first glass plate. [Figure 3c] This figure schematically shows a further embodiment in a cross-section perpendicular to the surface of the first glass plate. [Modes for carrying out the invention]

[0048] Figure 1 shows a recess 2 within the first glass plate 1, which can be formed by etching the first glass plate 1 after irradiation with a pulsed laser beam. In this case, the bottom 3 shows a concave depression having a substantially parabolic cross-section. Such concave depressions are formed at any position 12 where the laser beam is irradiated point by point. In this case, the bottom 3, along with the concave depression, is produced by irradiating the laser beam point by point at positions having intervals corresponding to the intervals between the center points of the depressions. By etching performed after irradiation, the region of the first glass plate 1 is removed between these positions. In this integrated embodiment, the laser beam irradiated onto the first surface 4 may be set to penetrate into a portion of the volume of the first glass plate 1, and / or the entire surface of the second surface 5 opposite to the first surface 4 may be coated with etching resist.

[0049] Figure 2 shows a top view of the glass plate 1, illustrating the recess 2. The wall of the recess 2 forms a strong optical contrast with respect to the bottom 3, so the wall is clearly depicted as the circumferential boundary of the bottom 3. For example, when particles such as living cells Z are illuminated to pass through the bottom 3, and especially when the cells Z are marked with a dye such as a fluorescent dye, they can be seen with good contrast against the bottom 3.

[0050] Figure 3 shows embodiments of each recess 2 integrally formed on the glass plate 1. This shows the irradiation of laser pulses onto the first surface 4 of the glass plate 1 at multiple locations, for example, spaced apart by 1 μm to 10 μm, thus forming locations where etching will create exactly one recess 2. In this case, etching at each position 12 of the bottom 3 of the recess 2 may form concave depressions as partial recesses 2'.

[0051] As shown in Figure 3a), the glass tip 11 protruding from the bottom 3 into the recess 2 perpendicular to the first surface 4 is formed during etching when at least three positions 12 to which laser pulses are irradiated are spaced relatively far apart, for example, at intervals of 20 μm. In this case, each position 12 irradiated with a laser pulse creates a concave depression or partial recess 2' in the bottom 3 during etching.

[0052] Figure 3b) shows that laser pulses irradiated to each position 13 and penetrating deeper into the thickness of the glass plate 1 form further recesses 14 during etching at that thickness, and these recesses 14 extend deeper into the glass plate 1 than the depressions at other positions 12 where the laser pulses irradiated into the glass plate 1 to a smaller depth. The penetration depth of the laser pulses into the glass plate 1 can be predetermined by setting the focal position and / or the intensity of the pulse energy of the laser pulses.

[0053] Figure 3c) shows that irradiation of adjacent positions 13 with laser pulses penetrating deeper into the glass plate 1 forms a further recess 14 during etching at that position, and the bottom 3 of the recess 14 extends within the thickness of the glass plate 1, deeper than the recesses formed by other positions 12 where the laser pulses are irradiated to a smaller depth within the glass plate 1 during etching.

[0054] Embodiment 1: A method for manufacturing a number of glass reactors, each formed as a recess (2) in a first glass plate (1) having an integral structure, 1. A step of irradiating the glass plate (1) with a laser pulse of a wavelength that penetrates the glass plate (1) at spaced positions (12) at each location on the glass plate (1) where the recesses (2) are to be formed as reactors, wherein the focal position of the laser pulse is set so that the focal point of the laser pulse does not extend over the entire thickness of the first glass plate (1), 2. The step of etching the glass plate (1) to form the recess (2), the step of etching the glass plate (1) for a sufficient time to form the recess (2) having a depth along the location, 3. A method by which etching of the glass plate (1) is completed and a concave recess is formed at the bottom of any of the positions (12) in the location, provided that the depth of the recess (2) is equal to the thickness of the first glass plate (2), and therefore the recess (2) has a bottom (3) integrally formed with the first glass plate (1).

[0055] Embodiment 2: The method according to Embodiment 1, characterized in that the depth of the recess (2) is at least 30 μm.

[0056] Embodiment 3: The method according to Embodiment 1 or 2, characterized in that the recess (2) has an aspect ratio of at least 2 of depth to diameter, as measured on the plane of the first surface (4).

[0057] Embodiment 4: The method according to any one of Embodiments 1 to 3, characterized in that a laser pulse is irradiated onto the first glass plate (1) at a plurality of equally spaced positions (12) where the recess (2) is to be formed.

[0058] Embodiment 5: The method according to any one of Embodiments 1 to 4, characterized in that, by setting the focal position of the laser pulse such that the focal point of the laser pulse extends only into the first thickness portion (20), the laser pulse is radiated to a smaller depth in a part of the position (12) into the first thickness portion (20) of the first glass plate (1), and by setting the focal position of the laser pulse such that the focal point of the laser pulse extends only into the second thickness portion (21), the laser pulse is radiated more deeply into the adjacent second thickness portion (21) of the first glass plate (1) in a part of the position (12), and during etching, the recess (2) extending across the first thickness portion (20) is formed, and a partial recess (2') extending across the second thickness portion (21) spaced apart from the partial wall (22) is formed.

[0059] Embodiment 6: The method according to any one of Embodiments 1 to 5, characterized in that the positions (12) are arranged at intervals of up to 10 μm, and at least three of the positions are spaced at least 20 μm apart from each other.

[0060] Embodiment 7: The method according to any one of Embodiments 1 to 6, characterized in that the laser pulse is irradiated onto the glass plate with intensity, and the laser pulse corrects the glass plate (1) to the focal position.

[0061] Embodiment 8: The method according to any one of Embodiments 1 to 7, characterized in that a laser beam is irradiated onto the location of the first glass plate (1) where the recess (2) is to be formed, along a path that is closed in the circumferential direction.

[0062] Embodiment 9: The method according to Embodiment 8, characterized in that the laser beam irradiated onto the circumferentially closed path is formed by adjacently irradiated laser beam pulses.

[0063] Embodiment 10: The method according to any one of embodiments 1 to 9, characterized in that at least one surface of the first glass plate (1) is coated with etching resist (10) before step 2.

[0064] Embodiment 11: The method according to Embodiment 10, characterized in that etching is performed adjacent to the surface coated with the etching resist (10) until an oblique surface (9) surrounding the recess is formed.

[0065] Embodiment 12: The method according to any one of Embodiments 1 to 11, characterized in that the conductive path is applied to the glass plate (1) and covers at least a portion of the inner wall of the recess (2).

[0066] Embodiment 13: A reactor formed as a glass recess, obtained particularly according to the method of any one of claims 1 to 12, wherein the reactor has a depth of the recess of a glass plate (1) of at least 30 μm and an aspect ratio of at least 2 of the depth to the diameter, measured in the plane of the first surface (4) of the glass plate (1), the recess (2) is surrounded by a wall, the end face of the wall is arranged in a common plane and forms the first surface (4) of the first glass plate (1), and the recess (2) is excluded from the first surface (4), in the reactor, The reactor is characterized in that the recess does not extend over the entire thickness of the glass plate (1), the bottom (3) of the recess is formed from the material of the glass plate (1), and at least three concave depressions are formed in the bottom of at least one of the recesses.

[0067] Embodiment 14: The reactor according to Embodiment 13, characterized in that the bottom portion (3) of the reactor is formed by adjacent recesses (2), and the recesses are adjacent to each other in a plane that is parallel to the first surface (4) and parallel to the plane of the second surface (5) opposite to the first surface.

[0068] Embodiment 15: The reactor according to Embodiment 13 or 14, characterized in that at least one further recess (14) is formed in the bottom (3) of the recess (2), and the recess extends to a greater depth within the first glass plate (1).

[0069] Embodiment 16: The reactor according to any one of embodiments 13 to 15, characterized in that the bottom portion (3) has at least one glass tip integrally formed from the first glass plate (1) for use as an optical waveguide for near-field illumination of the recess (2), the glass tip extending into the recess (2) perpendicular to the first surface (4) of the first glass plate (1).

[0070] Embodiment 17: A method for analyzing a sample, comprising the steps of supplying a reactor which is manufactured according to the method described in any one of Embodiments 1 to 12 or is a reactor described in any one of Embodiments 13 to 16; placing a cell-free or cell-containing sample in the reactor; adding at least one reagent to the reactor; and performing an optical measurement of the reactor.

[0071] Embodiment 18: The method according to Embodiment 17, characterized in that the sample is cellular and the cells are individually arranged in concave depressions. [Explanation of Symbols]

[0072] 1 glass plate 2 recesses 2' Partial recess 3 bottom 4. First surface 5. Second surface 7 Longitudinal central axis 8 Etching Resist 11 Glass tip 12. Position of the irradiated laser pulse 13. Position of the laser pulse that penetrated deeper. 14 Further recesses 20 Upper first thickness portion 21 Lower second thickness portion Z cell

Claims

1. A method for manufacturing a number of glass reactors, each formed as a recess (2) in a first glass plate (1) having a single-piece structure, 1. A laser pulse having a maximum pulse length of 100 ps and a wavelength that penetrates the glass plate (1) is irradiated point-like and perpendicularly at each location on the glass plate (1) where the recesses (2) are to be formed as reactors, at spaced-apart positions (12), wherein the laser is adjusted so that the laser beam does not collide with the glass plate between the locations, and the focal position of the laser pulse is set so that the focal point of the laser pulse does not extend over the entire thickness of the first glass plate (1), 2. The step of etching the glass plate (1) to form the recess (2), wherein the etching is wet etching, and the step of etching the glass plate (1) for a sufficient duration of time to form the recess (2) having a depth along the location, 3. A method for completing the etching of the glass plate (1) and forming a concave recess at the bottom of any of the positions (12) of the location, where the depth of the recess (2) is only a part of the thickness of the first glass plate (1), and therefore the recess (2) has a bottom (3) integrally formed with the first glass plate (1).

2. The method according to claim 1, characterized in that the depth of the recess (2) is at least 30 μm.

3. The method according to claim 1 or 2, characterized in that the recess (2) has an aspect ratio of at least 2 to the diameter of the depth as measured on the plane of the first surface (4).

4. The method according to any one of claims 1 to 3, characterized in that a laser pulse is irradiated onto the first glass plate (1) at a plurality of equally spaced positions (12) where the recess (2) is to be formed.

5. The method according to any one of claims 1 to 4, characterized in that, by setting the focal position of the laser pulse such that the focal point of the laser pulse extends only into the first thickness portion (20), the laser pulse is radiated to a smaller depth into the first thickness portion (20) of the first glass plate (1) in a part of the position (12), and by setting the focal position of the laser pulse such that the focal point of the laser pulse extends only into the second thickness portion (21), the laser pulse is radiated more deeply into the adjacent second thickness portion (21) of the first glass plate (1) in a part of the position (12), and during etching, the recess (2) extending across the first thickness portion (20) is formed, and a partial recess (2') extending across the second thickness portion (21) spaced apart from the partial wall (22) is formed.

6. The method according to any one of claims 1 to 5, characterized in that the positions (12) are arranged at intervals of up to 10 μm.

7. The method according to any one of claims 1 to 5, characterized in that at least three of the aforementioned positions are spaced at least 20 μm apart from each other.

8. The method according to any one of claims 1 to 7, characterized in that the laser pulse is irradiated onto the glass plate with intensity, and the laser pulse corrects the glass plate (1) to the focal position.

9. The method according to any one of claims 1 to 8, characterized in that a laser beam is irradiated onto the location of the first glass plate (1) where the recess (2) is to be formed, on a path that is closed in the circumferential direction.

10. The method according to claim 9, characterized in that the laser beam irradiated onto the circumferentially closed path is formed by adjacently irradiated laser beam pulses.

11. The method according to any one of claims 1 to 10, characterized in that at least one surface of the first glass plate (1) is coated with etching resist (10) before step 2.

12. The method according to claim 11, characterized in that etching is performed adjacent to the surface coated with the etching resist (10) until an oblique surface (9) surrounding the recess is formed.

13. The method according to any one of claims 1 to 12, characterized in that the conductive path is applied to the glass plate (1) and covers at least a portion of the inner wall of the recess (2).

14. A reactor formed as a recess made of glass, wherein the reactor has a depth of at least 30 μm in the glass plate (1) and an aspect ratio of at least 2 to the diameter of the depth as measured on the plane of the first surface (4) of the glass plate (1), the recess (2) is surrounded by a wall, the end face of the wall is arranged in a common plane and forms the first surface (4) of the glass plate (1), and the recess (2) is excluded from the first surface (4), in a reactor, The reactor is characterized in that the recess does not extend over the entire thickness of the glass plate (1), the bottom (3) of the recess is formed from the material of the glass plate (1), at least three concave depressions are formed in the bottom of at least one of the recesses, and the concave depressions have a parabolic or conical cross-section.

15. The reactor according to claim 14, characterized in that the bottom portion (3) of the reactor is formed by adjacent recesses (2), and is arranged adjacently in a plane that is parallel to the first surface (4) and parallel to the plane of the second surface (5) opposite to the first surface.

16. The reactor according to claim 14 or 15, characterized in that at least one further recess (14) is formed in the bottom (3) of the recess (2), and the recess extends to a greater depth within the glass plate (1).

17. The reactor according to any one of claims 14 to 16, characterized in that the bottom portion (3) has at least one glass tip integrally formed from the glass plate (1) for use as an optical waveguide for near-field illumination of the recess (2), the glass tip extending into the recess (2) perpendicular to the first surface (4) of the glass plate (1).

18. A method for analyzing a sample, comprising the steps of: supplying a reactor manufactured according to the method of any one of claims 1 to 13, or a reactor according to any one of claims 14 to 17; placing a cell-free or cell-containing sample in the reactor; adding at least one reagent to the reactor; and performing an optical measurement of the reactor.

19. The method according to claim 18, characterized in that the sample is cellular and the cells are individually arranged in concave depressions.

Images