Glass element and system for analyzing biological substance, and method for manufacturing the same

Abstract

To provide a glass element having microchannels which is usable for spatial detection and analysis of a biological substance.SOLUTION: There is provided a glass element (1) for storing and / or transferring a biological substance (60), particularly, a cell and / or a cell component of human being, an animal or a plant, particularly, protein and / or DNA and / or RNA, wherein the glass element (1) includes a plurality of microchannels (2), and the microchannels (2) connect one surface of the glass element (1) and the surface on a side opposite to the glass element (1), or terminate in a blind hole. The microchannels 2 are tapered toward the upper face from the lower face. Particularly, the microchannel (2) are formed into funnel shapes in at least a partial region.SELECTED DRAWING: Figure 6

Description

[Technical field]

[0001] The invention relates to a glass element with microchannels, which can be used in particular for the spatial detection and analysis of biological materials, such as human, animal or plant proteins, antibodies and / or cellular DNA, which are part of a cellular complex, such as a tissue sample or a cell culture. The invention also relates to a system for the analysis of biological materials, and to a method for the production of the glass element.

[0002] The aforementioned analyses are generally referred to as cell and / or DNA diagnostics. The glass elements according to the invention allow the spatial analysis of biological material, also referred to in English as "spatial diagnostics" or "spatial biology". The aforementioned invention allows the spatial analysis of cellular DNA and protein expression patterns, while retaining information related to cellular complexes, for example tissue samples or cell cultures. The invention also allows for single molecule analysis of biomarkers, for example in biological or medical samples, such as blood, serum, urine samples, cell supernatants, among others.

[0003] A glass element for separating DNA in a liquid medium is known from US20150122656. In this glass element, channels are introduced into a glass plate by drilling with a laser beam. The method described above is based on the introduction of a filament into the glass with successive pulses of an ultrashort laser. By defocusing the laser, a channel with a diameter of 1 μm is formed from the filament through the substrate. This is said to result in a particularly smooth inner channel wall that at most deviates negligibly from an ideal cylindrical shape.

[0004] For spatial analysis, it is desirable to place as many channels as possible as close together in the glass element. It is also desirable to adjust the diameter of the channels to obtain sufficient collection efficiency for the biological material. However, each channel represents a weakening of the glass element, and there must be sufficient glass volume between the channels to obtain a minimum sufficient stability to ensure efficient use of the glass element. Thus, there is a trade-off between collection efficiency and spatial resolution.

[0005] The invention solves this problem by means of a glass element according to the independent claims. The method according to the invention provides a corresponding glass element. Preferred embodiments are given in the dependent claims.

[0006] The invention comprises a glass element for containing and / or transporting biological material (60), in particular human, animal or plant cells and / or cell components, in particular proteins and / or DNA and / or RNA, comprising a plurality of microchannels, the microchannels preferably connecting one surface of the glass element with an opposite surface of the glass element or terminating in blind holes, the microchannels tapering from the lower surface towards the upper surface, the microchannels preferably being funnel-shaped at least in some areas.

[0007] The underside of the glass element is the side facing the biological material to be analyzed in this application. The glass element is usually a plate-shaped element, a so-called slide. The microchannel can connect one surface of the glass element with the opposite surface of the glass element, in particular the underside with the upper surface. In that case, the microchannel runs through. However, it is also possible to form the microchannel as a blind hole, in which case it does not reach one side, i.e. by definition the upper surface. The blind hole bottom is then located in the surface area and / or near the surface. The microchannel expands from the side of one surface of the glass element towards the opposite surface and is preferably formed in the shape of a funnel at least in some areas.

[0008] Accordingly, in an advantageous glass element it is provided that the diameter of the channel inlet of the microchannels on the lower surface is larger than the diameter of the channel outlet or channel end in the region of the upper surface.

[0009] This can be considered as a result of the widening of the microchannel. The channel has an opening on one side of the glass element, the diameter of which is larger than the diameter of the opening or blind hole end on the opposite side. Usually this is the main surface of the plate-shaped glass element. The lower side is defined in this application as the side facing the biological material to be analyzed, the upper side is the opposite side, facing the analysis unit. The widening of the channel is usually located on the lower side and therefore faces the biological material to be analyzed. Particularly advantageously, the microchannel has a circular or elliptical diameter. By microchannel in this context is meant that its diameter is in the range of a few micrometers to several hundred micrometers.

[0010] The widening of the microchannels at the lower surface of the glass element, and thus of its basic funnel shape, increases the efficiency of the capture of biological material and ensures the mechanical stability of the glass element by providing more glass material between the microchannels at the upper surface of the glass element. In this way, the openings of the channels at the lower surface of the glass element can be placed very close to each other, thereby achieving good spatial resolution while still providing good mechanical stability.

[0011] Also possible according to the invention is a double funnel, in which the widening of the microchannel is located on the upper and lower surfaces of the glass element, in the central region of the microchannel the microchannel is formed as if in a constriction and / or cylindrical shape, with the glass material being present between these regions and thus in the volume of the glass element.

[0012] An advantageous embodiment relates to a glass element in which the microchannels, at least in the region of the channel inlet or the channel outlet of the surface area of ​​the glass element, have a diameter of 5 μm to 200 μm, advantageously 7 μm to 130 μm, particularly advantageously 10 μm to 100 μm.

[0013] The selection of these diameters allows the biological material to enter the microchannels on one side of the glass element, typically the lower side as mentioned above, and be transported towards the opposite surface. The selection of the diameter contributes to or determines that only the biological material of interest, typically less than 1 μm in length, can enter the microchannels, while larger unwanted cells and / or cellular components cannot.

[0014] In an advantageous glass element it is provided that the channel inlets have a diameter of between 5 μm and 200 μm, preferably between 10 μm and 100 μm.

[0015] As mentioned above, this can be a conical region tapering towards the opposite surface, but it can also be followed by a cylindrical region of approximately constant diameter, or a region that expands towards the opposite surface, or it can be followed by a cylindrical region followed by an expanding region.

[0016] The opening angle of the funnel-shaped region of all said microchannels is preferably between 0.1° and 30°, particularly preferably between 2° and 18°. The thickness of the glass element is preferably between 0.1 mm and 3 mm.

[0017] The selection of specific aperture angles allows for efficient fabrication, especially using the methods described below, and also contributes to good, uninterrupted transport of the biological material to be analyzed from the channel inlet to the other surface.

[0018] By choosing the aperture angle, it is possible to concentrate the material to be analyzed in one channel or to separate the biological material between two or more channels at the upper surface of the glass element. The concentration is determined by the ratio of the squares of the channel diameters, also called the radius ratio (R 下 ) 2 / (R 上 ) 2 This increases the sensitivity of the analysis by fluorescence spectroscopy and allows targeted interaction with functional molecules on the channel walls. Concentrations with a radius ratio of more than 5 are particularly advantageous.

[0019] The method according to the invention, described below, allows the production of this wide range, but advantageously in particular of very thin thicknesses.

[0020] According to the above embodiment, an advantageous glass element is provided in which a funnel-shaped region in the region of the channel inlet is followed by a cylindrical or funnel-shaped region, in particular in the region of the channel outlet and / or the end of the blind hole.

[0021] The aforementioned features allow the glass element to have a very high density of microchannels, which is also called microchannel density and refers to the number of microchannels per unit area of ​​the glass element surface, e.g. the number of microchannels per square millimeter. Since one channel inlet is always associated with one channel outlet or blind hole end, the microchannel density of the upper surface of the glass element is equal to the microchannel density of the lower surface of the glass element. This is therefore a generalized microchannel density.

[0022] Particularly preferably, the glass element according to the invention has a lower limit of at least 20 / mm 2 or at least 50 / mm 2 , especially at least 400 / mm 2 , in particular at least 10000 / mm 2 The upper limit is in each case advantageously up to 16000 / mm 2 It is.

[0023] Particularly preferred is a glass element in which the channel wall comprises a structure with a plurality of rounded, dome-shaped recesses, which have a depth of less than 5 μm, in particular an extent of 5 to 20 μm, preferably a roughness of 50 nm to 1 μm, this value referring to the average roughness value Ra.

[0024] The inventors have surprisingly found that it is advantageous if the walls of the microchannels are as far as possible not smooth but structured. The aforementioned depths and roughnesses represent advantageous embodiments, whereby depth means the recess in a direction perpendicular to the axis of the channel and extent means the length of the recess parallel to the axis of the channel.

[0025] These structures can be efficiently manufactured using the ultrashort pulse laser manufacturing process described below and subsequent etching steps.

[0026] Such a dome-shaped recess provides the advantage that, when filling a gel into a microchannel, firstly, the gel can be filled well and similarly the gel adheres well to the channel.

[0027] Another special advantage of the channel wall structures, especially the dome-shaped recesses, is that they allow tuning of the electroosmotic flow (EOF), which is the charge-induced movement of a liquid, e.g. a buffer solution in a glass capillary, along a polar surface in an electric field.

[0028] The choice of roughness can tune the contribution of the EOF to the transport of biological material within the channel: a highly rough polar surface can slow down the EOF compared to a less rough surface, which can reverse the direction of transport of polar biological material within the glass element.

[0029] The choice of channel wall roughness and concomitant tunability of the opening angle allows selective closure of the channel by opposing EOF and applied electric fields.

[0030] In an advantageous glass element it is provided that the channel outlets form an outlet matrix in which it is possible to associate with each channel outlet the position of a channel inlet.

[0031] In particular this allows to perform a spatial analysis of the biological material, since in particular the positions of the channel outlets or blind holes form an outlet matrix in which the position of a channel inlet can be associated with each channel outlet.

[0032] In other words, this means that the position of the inlet matrix can be correlated with the position of the outlet matrix. In the simplest case, the center points of the channel inlets on the lower surface of the glass element at least substantially coincide with the position of the channel outlet or blind hole end on the upper surface of the glass element. This corresponds to an embodiment of a microchannel in which the axis of the channel runs vertically through the main surface of the glass element. However, it is also possible for the axis of the channel to run obliquely through the main surface of the glass element. However, even in this case, the position of the channel inlets can be correlated with the position of the channel outlet or blind hole end, so that by analyzing the biological material to be analyzed at the channel outlet or blind hole end, its position relative to the source of the inlet matrix and / or the channel inlet can be unambiguously determined.

[0033] Advantageously, the glass element comprises, at least in the region of the microchannels, a glass characterized by exhibiting a low intrinsic fluorescence in the wavelength range between 300 nm and 700 nm.

[0034] Intrinsic fluorescence is the illumination intensity x 10 -6 It is desirable to have a lower intrinsic fluorescence. Since the intrinsic fluorescence is small, the fluorescent light does not interfere with the optical analysis of biological materials. The appropriate range of intrinsic fluorescence is, for example, 10 times the illumination light intensity. -8 ~10 -6 It is.

[0035] Particularly suitable glass compositions are mentioned in the examples below. In general, glasses that are characterized by high chemical resistance and / or high water resistance are particularly suitable. Alkali-free glasses are particularly advantageous.

[0036] Particularly alkali-free glasses and borosilicate glasses may be suitable glass materials, in particular commercially available glasses under the trade names AF32, AF35, AS87, D263, D263T, B270, MEMPAX, Willow, G-Leaf, EN-A1, BDA-E.

[0037] According to one exemplary embodiment, the composition of the glass comprises the following components in weight percent: [Table 1]

[0038] According to a further exemplary embodiment, the composition of the glass comprises the following components: [Table 2]

[0039] In another embodiment, the composition of the glass comprises the following components: [Table 3]

[0040] A further suitable composition of the glass is given by: [Table 4]

[0041] According to a further embodiment, the composition of the glass comprises the following components: [Table 5]

[0042] For all the above mentioned glass compositions, coloring oxides such as Nd2O3, Fe2O3, CoO, NiO, V2O5, MnO2, CuO, Cr2O3 may be added as appropriate.

[0043] As a fining agent, 0 to 2 wt % of As2O3, Sb2O3, SnO2, SO3, Cl, F and / or CeO2 can be added, and the total amount of all compositions is 100 wt %.

[0044] It may also be advantageous, and is encompassed by the present invention, if the surface energy of the walls of the microchannel is adjusted so that the biological material to be analyzed is transported along the walls or adheres to the walls. The surface energy can be adjusted, for example, by a coating.

[0045] In an advantageous glass element, it is provided that at least the walls of the microchannels are covered with a coating which repels or to which the biological material to be analyzed adheres, in particular in order to optimize the surface energy at least in some areas, the coating advantageously comprises a polymer, a silane, an oligonucleotide, an antibody, a protein, an antigen or nitrocellulose, or an inorganic coating, in particular metal particles, in particular gold and / or iron, or polymer-based nano- or microparticles, or a combination of the aforementioned.

[0046] As mentioned above, it is possible for a coating to be present at only a portion of a microchannel, for example only at the channel entrance, or for different coatings to be present within a channel, particularly along the axis of the channel.

[0047] Coatings can typically have a monolayer thickness, be in the nm range or even completely fill the channel. In the simplest case, the coating is applied by a dip-coating process. Especially in the case of small channel diameters, coating using vacuum processes, e.g. chemical vapor deposition (CVD) or atomic layer deposition (ALD), is suitable. It can be advantageous to selectively apply the coating inside the channel, e.g. by applying a lift-off material outside the channel or selectively performing sputtering on the surface of the glass element.

[0048] In particular, the advantageous structures of the walls of the microchannels described above increase the surface area of ​​the channel walls, so that these structures interact particularly advantageously with coatings and / or fillings. In the case of adhesive coatings, more surface area is provided for the biological material to be analyzed. In the case of transporting coatings, the coatings suppress or prevent and / or at least contribute to clogging of the structures by biological material. In the case of fillings, the adhesion of the fillings in the channels can be ensured or at least improved.

[0049] In an advantageous glass element, the microchannels are filled with a gel, which advantageously has pores which, depending on the choice and / or composition of the gel, can have different diameters and thus can accommodate and / or transport biological materials of different sizes.

[0050] Particularly suitable within the meaning of the present invention are polyacrylamide gels and / or agarose gels. Agarose gels usually have larger pores than polyacrylamide gels and can be used in particular for the analysis of DNA and / or relatively large proteins. Polyacrylamide gels generally have smaller pores and are usually used for the analysis of relatively small proteins.

[0051] Gels, especially the aforementioned gels, can be used within the meaning of the present invention as carrier gels for electrophoretic processes, also commonly referred to as gel electrophoresis, which in particular allow different small molecules, especially DNA, RNA and proteins, to be separated from one another and analysed in a subsequent step, whereby the term gel electrophoresis generally includes the transport and / or migration of the analytes through the gel, without necessarily requiring separation.

[0052] Here, the molecules to be separated and / or transported advantageously move in a carrier gel present in the microchannel. This movement can be assisted by the application of an electric field, as described below. Depending on the size and charge and / or strength of the applied electromagnetic field, the molecules move at different distances and / or speeds.

[0053] In classical electrophoresis, molecules form characteristic banding patterns in microchannels, which can be visualized by staining and identified by associating their characteristic colors with the molecules.

[0054] However, in the case of glass elements according to the invention, in particular thin glass elements, it is further provided to use a gel and / or an alternative substance to transfer the substance to be analysed by electrophoresis from the lower surface of the glass element to the upper surface of the glass element, where the analysis is then carried out, for example by fluorescence spectroscopy or nanopore-based DNA sequencing, which are standard methods and will not be described in detail here.

[0055] Particularly advantageous are glass elements in which at least one DNA- or RNA-sensitive dye, in particular a plurality of dyes, is present in the region of the channel outlet or at the bottom of the blind hole.

[0056] When DNA and / or RNA from the biological material to be analyzed is transported through the microchannel, it reaches the outlet of the channel and / or the bottom of the blind hole if the microchannel is formed as a blind hole, where it can react with the DNA- or RNA-sensitive dye applied thereto, and the presence of the corresponding DNA or RNA strand is detected by its characteristic fluorescence, particularly its wavelength.

[0057] Advantageous dyes which can be used can be, for example, ethidium bromide, propidium iodide, crystal violet, 4',6-diamidino-2-phenylindole and / or 7-aminoactinomycin D.

[0058] In one advantageous configuration of the glass element, it is provided that in the region of the channel outlet or at the bottom of the blind hole, molecules for polymerase chain reaction, in particular DNA, primers, nucleotides and / or the enzyme DNA polymerase, are present. This embodiment is particularly suitable for single molecule analysis, in which the substance to be analyzed is first amplified in the microchannel and / or blind hole, as described herein. In particular, it is clear that it is also possible to introduce the aforementioned molecules into the microchannel and / or blind hole precisely when the substance to be analyzed is introduced into this microchannel and / or blind hole.

[0059] This embodiment has the particular advantage that the amplification of cells and / or DNA and / or RNA in the material to be analyzed is carried out in a very small volume in the microchannel (including blind holes). As a result, the time required for the necessary amplification can be reduced. Also, the analysis (including detection) of the material to be analyzed can be carried out in this small volume, resulting in good evaluability, especially good signal quality. In particular, the increased precision allows the identification of rare populations and the discrimination of subtle differences between cells. In this way, new biomarkers and signature molecules can be identified. Furthermore, the analysis of individual molecules and cells in a sample allows a statement to be made about the heterogeneity of molecules and cells in the sample. In particular, individual differences between patients can be visualized in order to develop personalized therapies based on specific properties of cells and molecules. Overall, this provides an efficient and reliable analysis with potential applications in personalized medicine.

[0060] The aforementioned glass elements are, so to speak, precursors of the system according to the invention, with which spatial and / or single molecule or single cell diagnostics can be carried out. As mentioned above, in a subsequent process, biological material can be analyzed and / or amplified on the glass elements or, for example, by DNA sequencing. In this way, gene and / or protein expression patterns and distribution patterns of DNA markers of individual cells can be determined simultaneously, with the location information being preserved within the cell complex or tissue sample.

[0061] A system for the analysis of biological material, in particular human or animal or plant cells and / or cell components, in particular proteins and / or DNA and / or RNA, comprises a glass element with a number of microchannels, in particular the glass element as described above, which serves in the operative state to accommodate and / or transport the biological material in or through the microchannels from the lower surface of the glass element to the region of the upper surface of the glass element, and which comprises an evaluation unit with which the biological material can be analyzed in the operative state. Within the meaning of the invention, the evaluation unit is assigned to or associated with the glass element.

[0062] The evaluation unit generally determines the presence of the biological material and / or its components transported through and / or present in the microchannel. In particular, the evaluation unit can comprise an electronic image sensor capable of capturing in particular the intrinsic fluorescence of the aforementioned dyes. Thus, the configuration of the microchannel as a blind hole is also included.

[0063] Thus, in an advantageous system, it is provided that at least one DNA or RNA sensitive dye is present in the region of the channel outlet (22) and / or in the blind hole (25), in particular at its bottom, and the color information of said dye can be captured, in particular spatially, by the evaluation unit.

[0064] For DNA analysis, a common DNA binding dye is, for example, ethidium bromide.

[0065] Advantageously, the system comprises, in an operational state, a sample support on which the biological material to be analyzed is placed, which is typically a sample plate or slide.

[0066] Particularly advantageous is a system in which the glass element is assigned a transport unit, by means of which biological material is introduced into the microchannels of the glass element in the operating state.

[0067] A transport unit is generally a unit that exerts a force on the biological material to be analyzed that at least assists its movement and / or transport into the microchannel of the glass element and towards the opposing surface.

[0068] Advantageously, the transport unit comprises a negative pressure unit which applies negative pressure to the outlet side of the glass element in an operating state and / or a unit which generates a potential gradient across the thickness of the glass element to introduce and / or transport the biological material to be analyzed through the microchannel by ion flow or an electric field.

[0069] Since DNA is essentially negatively charged due to its phosphate residues, it migrates to the anode during gel electrophoresis. Therefore, it is particularly advantageous to provide an anode on the upper surface of the preferred glass element or to assign the upper surface to the anode. If a cathode is present on or near the lower surface, application of a voltage creates an electric field that causes anions to migrate to the anode and cations to migrate to the cathode.

[0070] During DNA analysis it is of course possible to amplify DNA fragments beforehand and / or within the channels of the glass element, in particular by polymerase chain reaction.

[0071] It is of course possible and encompassed by the invention to provide the anode on the lower side of the glass element and the cathode on the upper side and / or to assign them respectively, depending on the biological material to be analyzed. Other arrangements are also possible and encompassed by the invention. For example, it may be sufficient to move the medium in which the biological material to be analyzed is present by a transport unit.

[0072] Also advantageously, the transport unit comprises a blade unit, which allows the biological material to be analyzed to be introduced into the microchannels, in particular into the blind holes, by spreading it out.

[0073] An advantageous system comprises as transport unit a microfluidic system, which introduces the biological material to be analysed into the blind holes and / or microchannels by means of a fluid flow.

[0074] Particularly advantageously, the system has a development in which the evaluation unit comprises a closed or nanoporous membrane on the outlet side of the microchannel, in particular made of a non-conductor or semiconductor or a combination of non-conductive, conductive and / or semi-conductive components, in particular glass, silicon, graphene and / or biomolecules, in particular lipid molecules, DNA origami structures or transmembrane proteins, by means of which the biological material can be analyzed in particular with regard to its sequence, binding behavior, size or optical properties, in particular fluorescence.

[0075] In an alternative configuration, the system comprises an evaluation unit on the upper surface of the microchannel in the form of a closed or nanoporous membrane, in particular made of a non-conducting material or a semiconductor, or a combination of non-conducting, conducting and / or semi-conducting components, in particular glass, silicon, graphene and / or biomolecules, in particular lipid molecules or transmembrane proteins, by means of which the biological material to be analysed is analysed, in particular for its sequence, binding behaviour, size or optical properties, in particular fluorescence.

[0076] As mentioned above, the microchannels can be filled with gels, especially to aid in transport. The gel itself can contain dyes that bind to the biological material to be analyzed. In the case of DNA analysis, DNA-binding dyes, such as ethidium bromide, are used. It is also possible for the sample to be in a sample buffer, which can contain dyes, such as crystal violet. It is also possible to stain the gel after electrophoresis with DNA-binding dyes, such as methylene blue, stainsall, or ethidium bromide. When using fluorescent dyes such as ethidium bromide, it is often necessary to illuminate the agarose gel with UV light and to equip the camera with a UV filter. RNA analysis is also possible.

[0077] In an advantageous system, in operation, it is provided to amplify biological materials in microchannels and / or blind holes, which is often referred to as digital PCR, in particular by providing molecules for polymerase chain reaction in the channel outlet region or at the bottom of the blind holes.

[0078] If the blind holes contain molecules for the polymerase chain reaction (PCR), then DNA or RNA molecules contained in the material to be analyzed can be amplified, e.g. DNA, primers, nucleotides and the enzyme DNA polymerase.

[0079] The temperature stability of the glass elements allows standard PCR up to approximately 100 °C in addition to PCR at room temperature, which is usually performed up to 40 °C. At these temperatures, in contrast to plastic, no adverse effects on the spatial information due to thermal expansion or misalignment of the blind holes with respect to one another cannot be expected. Furthermore, glass has the advantage over plastic of having a good thermal conductivity, which allows the heat to be distributed homogeneously within the glass element, ensuring a homogeneous PCR across the entire sample. Furthermore, glass has the advantage over plastic of having only a small intrinsic fluorescence, which allows a high signal intensity over the background.

[0080] As mentioned above, the outlet matrix of the microchannels on the upper surface of the glass element can be associated with the inlet matrix of the microchannels on the lower surface of the glass element, so that the system described above can determine the corresponding inlet positions of the biological material on the lower surface from the positions of the analysis results on the upper surface. In a manner of speaking, the positions of the microchannels of the glass element can be associated with the positions of the sources of the biological material in the sample. In this way, the glass element according to the invention and / or the system according to the invention can enable a spatial analysis of a sample of biological material. This is also called "Spatial Biology" or "Spatial Diagnostics".

[0081] The above-mentioned glass elements can be produced efficiently and in large quantities using the method according to the invention described below, which method comprises the steps of: - a method step of providing an ultrashort pulsed laser; - providing a glass element substrate; - irradiating a glass element substrate with a pulsed laser beam of an ultrashort pulse laser, wherein: - the wavelength of the laser beam and the material of the glass element substrate are matched to each other such that the workpiece is substantially transparent to the laser beam; the laser beam is focused in a focal region extending elongately in the beam direction, said focal region being at least partially present within the glass element body; - the intensity and focus of the laser beam are at a level such that the laser beam leaves a filamentary damage in the workpiece, introducing a plurality of such filamentary damages; - a method step of etching the filamentary damage to form a microchannel; Includes.

[0082] The glass element substrate can generally be a glass plate of the desired dimensions. A pulsed laser beam of an ultrashort pulsed laser is irradiated onto the glass element substrate, the wavelength of the laser beam and the material of the glass element substrate being matched to one another such that the workpiece is substantially transparent to the laser beam.

[0083] The laser beam is focused into a focal region that is elongated in the beam direction and that is at least partially located within the body of the glass element. For this purpose, various optical structures can be used, such as, for example, Bessel optics using axicons, chromatic aberration optics or spherical aberration optics.

[0084] The intensity and focus of the laser beam are set at such a level that the laser beam leaves filamentary damage in the workpiece and introduces multiple such filamentary damages, which represent thread-like damage in the glass and are etched into microchannels in a subsequent method step.

[0085] Filamentary damage is caused by nonlinear optical effects involving the self-focusing of a laser beam, which requires ultrashort laser pulses in the picosecond range.

[0086] An advantageous method provides that a series of laser pulses is used to generate the filamentary lesions.

[0087] In this so-called burst mode of operation, the laser energy is not emitted as a single pulse, but as a series of short successively emitted pulses, which together form a pulse packet, i.e. a so-called burst. Such a pulse packet usually has a slightly higher energy than a single pulse in the usual single-shot mode. However, the pulses of a burst themselves have significantly less energy than a single pulse. With regard to the pulses within a burst, it can be provided that the pulse energy is flexibly adjustable, in particular that the pulse energy is substantially constant or that the pulse energy increases or decreases.

[0088] A suitable laser source may be a neodymium-doped yttrium aluminum garnet laser with a wavelength of 1064 nanometers. This laser source may produce a raw beam with a (1 / e2) diameter of 12 mm, for example, and a biconvex lens with a focal length of 16 mm may be used as the optics. The raw beam may be produced using suitable beam shaping optics, for example a Galilean telescope.

[0089] The laser source can in particular be operated at a repetition rate of 1 kHz to 1000 kHz, advantageously 2 kHz to 100 kHz, particularly preferably 3 kHz to 200 kHz.

[0090] Here, the repetition rate and / or scan speed can be selected to achieve a desired distance between adjacent filamentary lesions.

[0091] Suitable pulse durations of the laser pulses may in particular be in the range of less than 100 picoseconds, advantageously less than 20 picoseconds.

[0092] Here, the typical power of the laser source is particularly preferably in the range of 20-300 watts. The laser energy is essentially below the ablation energy of the substrate. According to an advantageous development of the invention, a pulse energy of more than 400 microjoules is used in a burst, more preferably a total burst energy of more than 500 microjoules, to obtain filamentary damage.

[0093] When the ultrashort pulse laser is operated in burst mode, the repetition rate is the repetition rate of the burst emission. The pulse duration is substantially independent of whether the laser is operated in single pulse mode or burst mode. The pulses in one burst usually have a similar pulse length as the pulses in single pulse mode. The burst frequency is in the range of 15 MHz to 90 MHz, preferably in the range of 20 MHz to 85 MHz, and can be, for example, 50 MHz, and the number of pulses in a burst can be 1 to 10 pulses, for example, 6 pulses.

[0094] What is advantageous is a method in which the extended focal region includes a region having the highest energy followed by an expanded portion, the extended focal region being arranged such that the expanded portion is located in a surface region of the glass element substrate.

[0095] In this way, tapered microchannels can be produced in a particularly advantageous manner, as so-called filament-like pre-damage is generated by the damaged area, which results in an enlargement of the channel inlet in a subsequent etching step.

[0096] A particularly advantageous method involves preventing the filamentary damage from reaching at least one surface of the glass element substrate, and openings to this surface being made by a subsequent etching step.

[0097] This can be achieved by selecting the focal position and thus also the intensity of the laser beam. Since the creation of filamentary damage is a nonlinear effect, a local beam intensity must be achieved in order to cause damage to the glass as a whole; below this critical threshold, there is practically no damage to the glass, or at least not enough damage to form filaments. Only by the subsequent etching process does the filamentary damage expand and, so to speak, form a breakthrough through the corresponding surface of the glass element substrate. The inventors have found that this allows the matrix of channel outlets of the microchannels to be produced particularly precisely. Particularly precisely in this context means that the diameter of the channel outlets is small and / or that the channel outlets deviate at most only slightly from the ideal circular diameter.

[0098] In the simplest case, the center point of the channel inlet can be located above the center point of the channel outlet. The microchannel then passes through the glass element, so to speak, perpendicularly. However, it is also possible that the center points of the channel inlet and the center points of the channel outlet are offset from one another, in which case the microchannel passes through the glass element at an angle offset from the perpendicular to the glass element surface. Particularly suitable are angles of up to about 10° from this perpendicular direction, i.e. in the range of 0.1° to 10°. In particular, it may not be the case that the passage angle of the microchannel is the same everywhere in the glass element. By locally adjusting the passage angle, the channel matrix of the glass element can be adapted to the requirements.

[0099] The location of the widening of the channel opening is mainly due to the focus position. As mentioned above, the subsequent etching step will cause damage to the glass material to grow. The larger the damaged area, the larger the widening.

[0100] The shape of the elongated focus is utilized to tailor the expansion characteristics of the channel entrance such that the expansion region is followed by a region of highest energy when viewed along the axis of the beam, and the elongated focus region is positioned such that the expansion portion is located at a surface region of the glass element substrate.

[0101] During subsequent etching, this results in an enlarged channel inlet, in particular a funnel-shaped widening. The etching step is advantageously carried out in such a way that the microchannels have a diameter of 5 μm to 200 μm, preferably 7 μm to 130 μm, particularly preferably 10 μm to 100 μm, at least in the channel inlet region.

[0102] In one variant of the method, it is provided that the filamentary damage does not reach at least one surface of the glass element substrate, openings to this surface being made by a subsequent etching step.

[0103] This is because it is also very important to provide the most accurate possible geometry of the channel outlet, which may be located in particular in the cylindrical region of the microchannel. In order to adjust the geometry of the channel outlet, it has been found to be advantageous to ensure that the filamentary damage does not penetrate the surface of the glass element on which the channel outlet is located. In that case, the filamentary damage terminates, so to speak, below this surface. This is also adjusted by the corresponding focus position. In this case, the opening of the microchannel, i.e. the channel outlet, is formed by a subsequent etching step.

[0104] The surface of the glass element substrate can be treated, in particular by grinding and polishing, before and / or after etching. Grinding and polishing after the etching step has been found to be particularly advantageous, since a particularly flat surface can be achieved also around the channel inlets and / or channel outlets.

[0105] An advantageous method involves the introduction of recesses, which are hemispherical dome-shaped depressions, into the inner walls of the channels by etching.

[0106] This is especially true when slow etching processes are carried out, in particular with etching rates below 15 μm per hour.

[0107] The dome-shaped recess is assumed to be due to the structure that occurs when filamentary damage is introduced. A liquid etching solution is particularly advantageous as the etching medium. According to this embodiment, the etching is performed wet-chemically. This may also be advantageous for removing glass components from the surface during etching.

[0108] As etching solutions, both acidic and alkaline solutions can be used. As acidic etching media, in particular HF, HCl, H2SO4, ammonium bifluoride, HNO3 solutions or mixtures of these acids are suitable. As basic etching media, preferably KOH or NaOH alkaline solutions are included. With acidic etching solutions, usually higher removal rates can be achieved. However, basic solutions are preferred, especially if only slow removal is desired in any case.

[0109] Etching is preferably carried out in a temperature range between 40° C. and 150° C., particularly preferably between 50° C. and 120° C. In general, silicate glasses with a low alkali content are particularly suitable for the structuring according to the invention. If the alkali content is too high, etching becomes difficult. According to one development of the invention, it is therefore provided that the glass of the glass element is a silicate glass with an alkali oxide content of less than 17% by weight.

[0110] The glass element with microchannels described above can advantageously be used for the analysis of biological material, in particular human or animal or plant cells and / or cell components, in particular proteins and / or DNA.

[0111] As an alternative to spatial diagnostics, glass elements can be used for single molecule or single cell diagnostics due to their numerous blind holes. In this case, in particular liquid samples are introduced into the blind holes, for example by spreading with a blade process or the process mentioned above. Depending on the concentration selected, this ensures that none, only a few or just one molecule or cell to be analysed enters the blind hole. The molecules and / or cells to be analysed can then be selectively multiplied in the individual blind holes in order to carry out an analysis, in particular a fluorescence analysis.

[0112] In a particularly advantageous embodiment, it is provided to carry out specific tests on each of the blind holes or groups of blind holes in combination with a microfluidic cell that allows targeted control of the individual blind holes or groups of blind holes, which is particularly advantageous for determining the heterogeneity of samples and for medical diagnostics for carrying out a large number of different tests in a small space (highplex diagnostics).

[0113] The present invention will now be described in more detail with reference to the drawings. The drawings and related embodiments are examples, and the present invention is not limited thereto. In the drawings, the same reference numerals respectively indicate the same or corresponding elements. In addition, reference numerals shown in one drawing may also apply to other drawings, even if not shown. The drawings show schematic embodiment examples, and the proportions do not necessarily correspond to the proportions or dimensions of objects according to the present invention. [Brief description of the drawings]

[0114] [Figure 1] 1a to 1d are cross-sectional views of various embodiments of glass elements with enlarged microchannels. [Diagram 2] FIG. 1 shows a cross-sectional view of a portion of a glass element with enlarged microchannels. [Diagram 3] FIG. 1 shows an electron microscope photograph of a microchannel wall. [Figure 4] 4a and 4b are top views of the upper and lower surfaces of a glass element. [Diagram 5] FIG. 2 shows a micrograph of a cross section of a glass element. [Figure 6] FIG. 1 shows a system for spatial analysis of biological material. [Figure 7] FIG. 2 shows the layout of dyes on the top surface of a glass element. [Figure 8] FIG. 1 shows the layout of dyes in blind holes of a glass element. [Figure 9] FIG. 1 illustrates an apparatus for laser machining of glass elements in preparation for subsequent etching. [Figure 10] 10a and 10b are schematic diagrams illustrating the effect of structures on the microchannel walls.

[0115] Figure 1a shows an embodiment of a glass element (1) with a number of microchannels (2). In this example, the microchannels connect the upper (O) and lower (U) faces of the glass element and pass through the glass element. As mentioned above, the upper face generally refers to the surface or face of the glass element that faces away from the sample or on which the analysis is carried out. The lower face is usually the surface or face of the glass element that faces the sample, i.e. the biological material to be analyzed. In the terminology used herein, the channel inlets (21) are located in the area of ​​the lower face (U) and the channel outlets (22) are located in the area of ​​the upper face (O).

[0116] In the present example of Fig. 1a, the microchannel tapers, in this example continuously, from the lower surface (U) to the upper surface (O). In general, the widening in at least some areas is referred to herein as funnel-shaped. In the example shown, the taper is continuous. The profile of the microchannel (2) can also be referred to here as a truncated cone. The tapered area extends here over the entire thickness (s) of the glass element.

[0117] The thickness or material thickness (s) of the glass element after processing can in particular be 0.1 mm to 3 mm, as mentioned above. The illustrated glass element (1) has substantially plane-parallel surfaces (O) and (U), as are usually the side surfaces. The glass element (1) is advantageously a slide.

[0118] 1b shows a similar embodiment in which the taper of the microchannels is more pronounced: the microchannels (2) taper towards the upper surface (O) and / or are funnel-shaped at least in some areas, so that, within the meaning of the invention, the diameter (dU) of the channel inlet (21) at the lower surface (U) is larger than the diameter (dO) of the channel outlet (22) at the upper surface (O) of the glass element (1).

[0119] FIG. 1c shows a further embodiment in which the profile of the microchannel consists of a tapered truncated cone region in the region of the lower surface (U) and a cylindrical region in the region of the upper surface (O). The overall shape can be called funnel-shaped as well. As mentioned above, the center points of the channel outlets (22) determine the pitch (P). The smaller the pitch, the higher the density of the microchannels, so the aim is to have a pitch as small as possible. The pitch can be between 3 and 70 μm, as mentioned above. As shown in this example, the center point of the channel inlet (21) is substantially above the center point of the channel outlet (22). In other words, these two center points are substantially on an axis that passes perpendicularly through the surfaces (O) and (U) of the glass element.

[0120] Figure 1d shows an embodiment in which the microchannel (2) tapers from both the lower (U) and upper (O) faces towards the bulk of the glass material. In the shape shown as an example here, this results in an hourglass or X-shape, which here is essentially two inverted truncated cones stacked on top of each other with their short sides touching.

[0121] Figure 2 shows a part of the glass element (1) according to Figure 1b. The opening angle α is entered, which should be interpreted as the angle between the channel walls of the expanding region of the microchannel (2). Advantageously, the opening angle α is between 0.1° and 30°, particularly advantageously between 2° and 18°, it being understood that the upper and lower limits of this interval can of course be variable.

[0122] As explained in detail above, this tapered microchannel makes it possible to achieve the contradictory objectives of, on the one hand, aiming for the smallest possible pitch in order to increase the resolution of the analysis, leading to high detection sensitivity and thus a large diameter of the channel inlet, and, on the other hand, the need to ensure the mechanical stability of the glass elements in order to be able to use them efficiently, since a particularly large amount of material remains in the non-tapered areas.

[0123] Figure 3 shows an electron microscope photograph of the walls of the microchannels (2) of a glass element (1) produced by the inventors according to the invention, in particular a portion of the region (40) of the microchannel as shown in Figure 1c. As already mentioned herein, the lateral dimension or diameter of the channel (2) is less than 200 micrometers, in this example about 14 micrometers. Without being limited to the examples described here, diameters in the range of 5 micrometers to 200 micrometers are preferred.

[0124] The dome-shaped or rounded cap-shaped recesses (7) caused by the etching of the filamentary damage introduced into the glass element by the ultrashort pulsed laser beam are clearly visible. The depth of the dome-shaped recesses (7) is typically less than 5 μm and the lateral dimensions are typically 5-20 μm.

[0125] As a result of the filament damage etching process, the dome-shaped recesses (7) are adjacent to one another, and the concave curves of the recesses (7) are tangent to one another to form ridges (70). Moreover, when viewed from above the recesses (7), it can be seen that the ridges (70) form the polygonal boundary (71) of the recesses (7), where the average number of corners (72) of the boundary (71) of the recesses (7) is advantageously also less than 8, advantageously less than 7. The latter feature arises when the area occupied by the majority of the dome-shaped recesses is convex in the mathematical sense.

[0126] The ridges (70) of the channel 2 shown in Fig. 3 are very narrow and there is no discernible area where the concave curvatures of the recesses (7) meet each other through a convex curvature area at the ridges (70).According to one development of the invention, the structure of the channel (2) can therefore also be described as having an area percentage of convex areas in the channel (2) of less than 5%, advantageously less than 2%.

[0127] Figure 4a shows a schematic top view of the upper (O) and lower (U) faces of a glass element (1). The diameters of the channel inlets (21) and the channel outlets (22) are substantially equal. Such a pattern can be envisaged, for example, in the embodiment according to figure 1d.

[0128] Figure 4b shows a similar top view, but with the diameter of the channel inlet (21) being smaller than the diameter of the channel outlet (22).Such a pattern corresponds for example to the embodiment according to figures 1a to 1c.

[0129] As shown in Figures 4a and 4b, the channel inlets and outlets form a matrix. The positions of the channel inlets are correlated to the positions of the channel outlets, so that if the position of the channel outlet is known, a spatial measurement of an event, e.g., the presence of DNA or a particular biological material, can be performed at the channel inlet side, and therefore at the sample side.

[0130] FIG. 5 shows a photograph taken by an optical microscope of a cross section of a glass element (1) produced by the inventors, which corresponds substantially to the embodiment according to FIG. 1c. The funnel-shaped expanded area is followed by a cylindrical area of ​​the microchannel (2). The diameter of the channel inlet (21) on the lower surface of the glass element (1) is larger than the diameter of the channel outlet (22) on the upper surface. In this example, glass D263 from SCHOTT was used. The invention is not limited to a specific type of glass. In other examples, borosilicate and / or alkali-free glasses were produced. Borosilicate glasses in particular are frequently used in laboratories due to their chemical resistance. As already mentioned, glasses that show only a little intrinsic fluorescence in the visible region of the spectrum are particularly advantageous.

[0131] Figure 6 shows an exemplary spatial measurement system (10) for biological material (60). The biological material to be analyzed is here present on a sample support (62). A glass element (1) with the aforementioned microchannels (2) is in contact with the biological material (60). Of course, it is also possible to bring the glass element (1) into contact with the biological material without preparing the biological material on a support. The glass element can be in direct contact with, for example, tissue or cell cultures.

[0132] In the microchannel (2) there may be a gel, in particular the aforementioned gel, which is not shown in FIG. 6. At the top side of the glass element (1), in the region of the channel outlet (22), at least one DNA or RNA sensitive dye (610) is applied. When the biological material (60) passes through the microchannel (2), in particular by means of the gel, it comes into contact with the dye (610) at the top side, the typical color of which may then indicate the presence, and in some cases the presence of a particular DNA strand. As mentioned above, the dye is usually illuminated with an excitation light, after which the typical fluorescence of the dye is measured with the aid of an evaluation unit, which is not shown in this figure for the sake of clarity. As also mentioned above, this may in particular be a digital camera.

[0133] Advantageously, different channel outlets (22) may be provided with different dyes (610) so that different DNA or DNA strands can be identified during the analysis of biological material.

[0134] The top view shows the patterns that allow spatial estimation of the presence of certain biological materials, particularly DNA, in the illustrated system (10). Everything said about DNA also applies to RNA.

[0135] The transport of the biological material (60) into or through the microchannel (2) can be assisted by the presence of a transport unit (65). In the example of FIG. 6, this is a unit that generally uses an electrical potential for the transport of the biological material. Shown is a capacitor unit in which a voltage source (V) is connected to electrodes (651, 652). In this example, the electrodes are arranged on the underside of the sample support (62) and on a non-conductive plate (653), in particular a glass plate or plastic. The electrodes thus form a plate capacitor, between whose plates the biological material (60) and the glass element (1) are present. When a voltage is applied, an electric field is generated, which pulls or pushes the biological material and / or the surrounding medium towards the electrode (651).

[0136] If a non-conductive plate (653) is brought into contact with the upper surface (O) of the glass element, this non-conductive plate (653) can also act as a closure of the microchannel (2), so to speak creating a blind hole, so that the advancing biological material does not remove the dye from the surface of the glass element.

[0137] The electrodes (651, 652) can be efficiently applied by coating, in particular by sputtering.

[0138] Other arrangements are possible and are encompassed by the present invention, in particular the use of pressure and / or negative pressure instead of electromotive force to transport biological material.

[0139] Figure 7 shows a top view of a glass element (1) with sensitive dyes (610, 611, 612, 613, 614) on its upper surface (O) in particular in a system according to figure 6. The illustrated patterns show diagrammatically possible analytical results.

[0140] FIG. 8 shows again for clarity an embodiment of a glass element with microchannels (2) formed as blind holes (25). The aforementioned dyes (610, 611) are applied to the blind hole bottoms. The electrodes (651) of the electrical transport unit (65) can simply be applied to the surface of the glass element. In order to be able to detect the dye fluorescence through the glass material located above the blind hole bottoms, the electrodes are in this case in particular transparent electrodes, which can be produced, for example, as an ITO coating. It is also possible for the electrodes to be formed as rasters and / or grids, in particular with voids above the bottoms of the corresponding blind holes (25).

[0141] 9 shows an embodiment of a laser processing device (20) in which a filamentary damage (32) can be introduced into a glass element substrate (1) and then a microchannel can be introduced at the location of the filamentary damage (32) by an etching process. The device (20) comprises an ultrashort pulsed laser (300) to which a focusing optical system (230) is already connected, and a positioning unit (170). The positioning unit (170) can laterally position the impact point (73) of the laser beam (270) of the ultrashort pulsed laser (300) on the surface (O) of the plate-shaped glass element (1) to be processed. In the illustrated example, the positioning unit (170) comprises an XY table, on which the underside (U) of the glass element (1) is placed. However, alternatively or additionally, the optics can be movable to move the laser beam (270), thereby moving the impact point (73) of the laser beam (270) while keeping the glass element (1) fixed.

[0142] Figures 10a and 10b show schematics of the effect of structures on the microchannel walls. Figure 10a shows a smooth channel wall, whereas Figure 10b shows a structured channel wall with roughness Ra. The channel wall has an opening angle. The structures can have the shape of dome-like recesses. EOF refers to electroosmotic flow, which, as previously mentioned, is the charge-induced movement of liquid along a polar surface in an electric field. A high roughness of the polar surface, as shown in Figure 10b, can slow down the EOF compared to a low roughness surface, as shown in Figure 10a, and can reverse the direction of movement of polar biomaterials in the glass element.

[0143] The focusing optics (230) focuses the laser beam (270) into a focus that is elongated in the beam direction, i.e., elongated transversely, in particular perpendicularly, to the irradiation surface (O). Such a focus can be generated, for example, by means of a conical lens (so-called axicon) or a lens with a large spherical and / or chromatic aberration. The positioning unit (170) and the ultrashort pulse laser (300) are controlled, preferably by a program-equipped computer unit (150). In this way, a predefined pattern of filamentary damages (32) distributed transversely along the surface (O) can be generated, in particular by reading position data advantageously from a file or via a network. The shape of the filamentary damages (32) can be adjusted by the position of the focus, such that a local widening of the microchannels (2) generated by subsequent etching occurs.

[0144] According to one embodiment, the following parameters can be used for the laser beam: the wavelength of the laser beam is 1064 nm, which is typical for a YAG laser; a laser beam with a raw beam diameter of 12 mm is generated, which is focused with an optical system in the form of a biconvex lens with a focal length of 16 mm; the pulse duration of the ultrashort pulse laser is less than 20 ps, ​​preferably around 10 ps; the pulses are emitted in bursts of 2 or more, preferably 4 or more pulses; the burst frequency is between 12 and 48 ns, advantageously around 20 ns, the pulse energy is at least 200 microjoules, and accordingly the burst energy is at least 400 microjoules.

[0145] Then, after introducing one or in particular several filamentary damages (32), the glass element (1) is removed and stored in an etching bath in which an etching process removes the glass along the filamentary damages (32), thereby introducing a microchannel (2) into the glass element 1 at the position of each such damage (32).

[0146] Basic etching baths with a pH value >12 are preferred, for example KOH solutions of >4 mol / l, preferably >5 mol / l, particularly preferably >6 mol / l but <30 mol / l. According to one embodiment of the invention, the etching is carried out at an etching bath temperature of >70° C., preferably >80° C., particularly preferably >90° C., regardless of the etching medium used.

[0147] The glass elements and systems according to the invention have the advantage that they allow spatial analysis of biological material. The glass elements can be efficiently manufactured and consist of materials commonly used in laboratory environments. After use, the glass can be remelted, thus allowing recycling of the raw materials.

Claims

1. A glass element (1) for containing and / or transporting biological material (60), in particular human, animal or plant cells and / or cell components, in particular proteins and / or DNA and / or RNA, comprising: a plurality of microchannels (2) preferably connecting one surface (U, O) of the glass element (1) with an opposite surface (O, U) of the glass element (1) or terminating in a blind hole (25), The microchannel (2) is tapered from the lower surface (U) to the upper surface (O), Preferably, the microchannel (2) is formed in the shape of a funnel in at least some areas of the glass element (1).

2. 2. The glass element (1) according to claim 1, wherein the diameter (dU) of the channel inlet (21) of the microchannels in the lower surface (U) is larger than the diameter (dO) of the channel outlet (22) or channel end (25) in the region of the upper surface (O).

3. 3. The glass element (1) according to claim 1 or 2, wherein the microchannel (2) has a diameter (dU, dO) of 5 μm to 200 μm, preferably 7 μm to 130 μm, particularly preferably 10 μm to 100 μm, at least in the region of the channel inlet (21) or the channel outlet (22) or the channel end (25) of the glass element (1), and preferably has an opening angle (α) of 0.1° to 30°, particularly preferably 2° to 18°, of the funnel-shaped region.

4. 4. The glass element (1) according to claim 1, wherein the funnel-shaped region in the region of the channel inlet (21) is followed by a cylindrical or funnel-shaped region, in particular in the region of the channel outlet (22) or the end of the blind hole (25).

5. The density of the microchannels is at least 20 / mm ２ , in particular at least 400 / mm ２ , in particular at least 10000 / mm ２ In particular, in both cases, the maximum ２ 5. The glass element (1) according to claim 1, wherein

6. 6. The glass element (1) according to claim 1, wherein the channel wall of the microchannel (2) has a structure with a plurality of rounded dome-shaped recesses (7), the recesses (7) having a depth, in particular less than 5 μm.

7. 7. The glass element (1) according to any one of the preceding claims, wherein at least one DNA or RNA sensitive dye (610, 611, 612, 613, 614) is present in the region of the channel outlet (22) or at the bottom of the blind hole (25).

8. 8. The glass element (1) according to claim 1, wherein molecules for polymerase chain reaction, in particular DNA, primers, nucleotides and / or the enzyme DNA polymerase, are present in the region of the channel outlet (22) or at the bottom of the blind hole (25).

9. A system (10) for the analysis of biological material (60), in particular human, animal or plant cells and / or cell components, in particular proteins and / or DNA and / or RNA, said system (10) comprising a glass element (1) with a plurality of microchannels (2), in particular a glass element according to any one of claims 1 to 8, said glass element (1) serves, in an operating state, to contain and / or transport said biological material (60) in or through said microchannel (2) from said lower surface (U) of said glass element to the region of said upper surface (O) of said glass element, an evaluation unit capable of analyzing the biological material in an operational state, System (10).

10. The system (10) according to claim 9, wherein at least one DNA or RNA sensitive dye (610, 611, 612, 613, 614) is present in the region of the channel outlet (22) or at the bottom of the blind hole (25), and the color information of the dye can be grasped, in particular spatially, by the evaluation unit.

11. 11. The system (10) according to claim 9 or 10, wherein the glass element (2) is assigned a transport unit (65) by means of which, in an operating state, the biological material (60) to be analyzed is introduced into and / or transported through the microchannel (2) of the glass element (1).

12. The transport unit (65) a negative pressure unit which applies a negative pressure to the outlet side of the glass element in an active state, and / or a unit for generating an electric potential gradient, in particular across the thickness of the glass element, in order to introduce the biological material (60) to be analyzed into and / or through the microchannel (2) by means of a flow or an electric field, and / or A blade unit that introduces the biological material to be analyzed into the blind hole by spreading it. The system (10) of claim 11, comprising:

13. A system (10) according to any one of claims 9 to 12, in which in an operating state biological substances are amplified in the microchannel (2) and / or the blind hole (25), in particular in the region of the channel outlet (22) or at the bottom of the blind hole (25) molecules for polymerase chain reaction are present.

14. 14. The system (10) according to any one of claims 9 to 13, wherein the evaluation unit comprises a closed or nanoporous membrane on the outlet side of the microchannel (2), in particular made of a non-conductive material or a semiconductor, or a combination of non-conductive, conductive and / or semi-conductive components, in particular glass, silicon, graphene and / or biomolecules, in particular lipid molecules or transmembrane proteins.

15. A method for producing a glass element (1), in particular a glass element (1) according to any one of claims 1 to 7, said glass element (1) comprising a plurality of microchannels (2) connecting one surface (U, O) of said glass element with an opposite surface (O, U) of said glass element, said method comprising the steps of: The method steps of providing an ultrashort pulse laser (300); A method step of providing a glass element substrate (1), A step of irradiating the glass element substrate (1) with a pulsed laser beam (270) of the ultrashort pulsed laser (300), wherein: the wavelength of the laser beam (270) and the material of the glass element substrate (1) are matched to each other such that the workpiece is substantially transparent to the laser beam (270); the laser beam (270) is focused in a focal region (73) extending elongately in the beam direction, the focal region (73) being at least partially located within the glass element body (1); the intensity and focus of the laser beam (270) are at a level such that the laser beam (270) leaves a filament-like damage (32) in the workpiece, introducing a plurality of such filament-like damages (32); a method step of etching the filamentary damage (32) to form a microchannel (2); A method comprising:

16. 16. The method according to claim 15, wherein the filamentary damage (32) does not reach at least one surface (O) of the glass element substrate (1), and openings to said surface (O) are made by a subsequent etching step.

17. 17. The method according to claim 15 or 16, wherein the etching introduces a dome-shaped recess (7) into the inner channel wall of the microchannel (2).

Images