Glass elements and systems for analyzing biomaterials, and methods 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 present invention relates to a glass element equipped with a microchannel, which can be used particularly for the spatial detection and analysis of biomaterials. Such biomaterials include, for example, human, animal, or plant proteins, antibodies, and / or cellular DNA, which are part of a cellular complex, such as a tissue sample or cell culture. The present invention also relates to a biomaterial analysis system and a method for manufacturing the glass element.

[0002] The aforementioned analysis is generally referred to as cellular and / or DNA diagnostics. The glass element according to the present invention enables the spatial analysis of biomolecules, also known in English as "spatial diagnostics" or "spatial biology." The present invention allows for the spatial analysis of DNA and protein expression patterns in cells, while retaining information related to cell complexes such as tissue samples and cell cultures. Furthermore, the present invention also enables single-molecule analysis of biomarkers in biological and medical samples, particularly blood, serum, urine samples, and cell supernatants.

[0003] A glass element for separating DNA in a liquid medium is known from U.S. Patent Application Publication No. 20150122656. In this glass element, channels are introduced into a glass plate by perforation using a laser beam. The aforementioned method is based on introducing a filament into the glass using continuous pulses of an ultrashort pulse laser. By shifting the focus of the laser, a channel with a diameter of 1 μm is formed that penetrates the substrate from the filament. It is stated that this creates a particularly smooth channel wall that deviates at most negligibly from an ideal cylindrical shape.

[0004] For spatial analysis, it is desirable to place as many channels as possible within the glass element and as close together as possible. It is also desirable to adjust the channel diameter to obtain sufficient collection efficiency for biological materials. However, each channel weakens the glass element, and sufficient glass volume must exist between channels to obtain the minimum sufficient stability that guarantees the efficient use of the glass element. Therefore, collection efficiency and spatial resolution are inversely related.

[0005] The present invention solves this problem with a glass element according to an independent claim. The method according to the present invention provides a corresponding glass element. Preferred embodiments are shown in the dependent claims.

[0006] The present invention relates to a glass element for containing and / or transporting biomaterials (60), particularly human, animal or plant cells and / or cellular components, particularly proteins and / or DNA and / or RNA, wherein the glass element comprises a plurality of microchannels, the microchannels preferably connecting one surface of the glass element to the opposite surface of the glass element or terminating in blind holes, and the microchannels tapering from the bottom to the top surface. The microchannels are preferably funnel-shaped in at least some areas.

[0007] In this application, the bottom surface of the glass element is the surface facing the biological material being analyzed. The glass element is usually a plate-shaped element, a so-called slide. A microchannel can connect one surface of the glass element to the opposite surface, and in particular, it can connect the bottom surface to the top surface. In this case, the microchannel is through. However, it is also possible to form the microchannel as a blind hole, in which case the microchannel does not reach one surface, i.e., the top surface by definition. In this case, the bottom of the blind hole is located in the surface region and / or near the surface. The microchannel expands from one surface of the glass element toward the opposite surface, and is preferably funnel-shaped in at least a portion of the region.

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

[0009] This can be considered a result of the expansion of the microchannel. The channel has an opening on one face of the glass element, the diameter of which is larger than the diameter of the opening or blind hole end on the opposite face. Typically this is the main surface of the plate-shaped glass element. Its lower surface is defined as the surface facing the biomaterial being analyzed in this application, and its upper surface is the opposite surface facing the analysis unit. The expanded portion of the channel is usually located on the lower surface and is therefore facing the biomaterial being analyzed. Particularly advantageous, the microchannel has a circular or elliptical diameter. As used herein, a microchannel means a diameter in the range of several micrometers to several hundred micrometers.

[0010] By expanding the microchannels on the underside of the glass element, and consequently expanding its basic funnel shape, the collection efficiency of biological materials is improved. Furthermore, the amount of glass material present between the microchannels on the upper surface of the glass element increases, thereby ensuring the mechanical stability of the glass element. In this way, the openings of the channels on the underside of the glass element can be positioned very close to each other, thereby achieving good spatial resolution while still providing good mechanical stability.

[0011] Furthermore, a double funnel is also possible in this invention, in which case the enlarged portions of the microchannel are located on the upper and lower surfaces of the glass element. In the central region of the microchannel, the microchannel is formed in a constricted and / or cylindrical shape, and sufficient glass material is present between these regions and, consequently, within the volume of the glass element.

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

[0013] By selecting these diameters, biomaterials can enter the microchannel on one side of the glass element, usually the bottom side as described above, and be transported toward the opposite side. The selection of diameters contributes to, or determines, that only biomaterials of the analyte, typically less than 1 μm in length, can enter the microchannel, preventing larger, unwanted cells and / or cellular components from entering.

[0014] An advantageous glass element is provided that the channel entrance has a diameter of 5 μm to 200 μm, preferably 10 μm to 100 μm.

[0015] As mentioned above, it is possible to create a conical region that tapers towards the opposite surface, but it is also possible to provide a cylindrical region with a nearly constant diameter following the expanding region, or a region that expands towards the opposite surface. Furthermore, it is also possible to provide an expanding region following the cylindrical region.

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

[0017] By selecting a specified opening angle, efficient manufacturing becomes possible, particularly using the method described later. These values ​​also contribute to the smooth and efficient transport of the biomaterial to be analyzed from the channel inlet to the other surface.

[0018] By selecting the aperture angle, it is possible to concentrate the substance to be analyzed within a single channel or to separate biological material between two or more channels on the upper surface of the glass element. The degree of concentration is the ratio of the squares of the channel diameters (R), also known as the radius ratio. 下 ) 2 / (R 上 ) 2 This is proportional to the radius ratio, increasing the sensitivity of fluorescence spectroscopy analysis and enabling targeted interactions with functional molecules on the channel wall. A concentration of more than 5 radius ratios is particularly advantageous.

[0019] The method according to the present invention, described later, enables manufacturing over this wide range, but advantageously, it allows manufacturing at particularly thin thicknesses.

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

[0021] Due to the aforementioned features, it becomes possible for the glass element to have a very high density of microchannels. The density of microchannels, also referred to as the microchannel density, indicates the number of microchannels per unit area of the glass element surface, for example, the number of microchannels per square millimeter. Since one channel inlet is always associated with one channel outlet or the end of a blind hole, the microchannel density on the upper surface of the glass element is equal to the microchannel density on the lower surface of the glass element. Therefore, this can be said to be the general microchannel density.

[0022] Particularly advantageously, the glass element according to the invention has a microchannel density of at least 20 / mm 2 or at least 50 / mm 2 , particularly at least 400 / mm 2 , particularly at least 10000 / mm 2 . In any case, the upper limit is advantageously at most 16000 / mm 2 .

[0023] Particularly advantageous is a glass element in which the channel walls include a structural part with a plurality of rounded dome-shaped recesses, these recesses having a depth particularly less than 5 μm, a spread particularly of 5 - 20 μm, and an advantageous roughness of 50 nm - 1 μm. This value refers to the average roughness value Ra.

[0024] The inventors have found that it is advantageous for the walls of the microchannels to have a structural part rather than being as smooth as possible. The aforementioned depth and roughness represent advantageous embodiments. Here, the depth means the recess in the direction perpendicular to the axis of the channel, and the spread means the length of the recess parallel to the axis of the channel.

[0025] These structural parts can be efficiently manufactured by the manufacturing process using an ultrashort pulse laser and the subsequent etching process described below.

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

[0027] Another special advantage of the structural part of the channel wall, particularly the dome-shaped recess, is that it can adjust the electroosmotic flow (EOF). Electroosmotic flow is the charge-induced movement of a liquid along a polar surface in an electric field, and this liquid is, for example, a buffer solution in a glass capillary.

[0028] By selecting the roughness, the contribution of EOF to the movement of biomolecules in the channel can be adjusted. When the roughness of the polar surface is large, the EOF velocity decreases compared to a surface with small roughness, and the moving direction of polar biomolecules in the glass element may be reversed.

[0029] By selecting the roughness of the channel wall and simultaneously making the opening angle adjustable, selective closure of the channel by the reverse EOF and the applied electric field becomes possible.

[0030] In an advantageous glass element, it is provided that the channel exits form an exit matrix, and in this exit matrix, the positions of the channel entrances can be associated with each channel exit.

[0031] Particularly, this makes it possible to perform spatial analysis of biomolecules. This is especially because the positions of the channel exits or blind holes form an exit matrix, and in this exit matrix, the positions of the channel entrances can be associated with each channel exit.

[0032] In other words, this means that the position of the inlet matrix can be mapped to the position of the outlet matrix. In the simplest case, the center point of the channel inlet on the bottom surface of the glass element coincides at least substantially with the position of the channel outlet or blind hole end on the top surface of the glass element. This corresponds to an embodiment of a microchannel in which the channel axis extends perpendicularly through the main surface of the glass element. However, it is also possible for the channel axis to extend obliquely through the main surface of the glass element. Even in this case, however, the position of the channel inlet can be mapped to the position of the channel outlet or blind hole end, so that by analyzing the biomaterial under analysis at the channel outlet or blind hole end, the position of the inlet matrix and / or channel inlet relative to the source can be clearly determined.

[0033] Advantageously, the glass element includes, at least in the microchannel region, a glass characterized by exhibiting slight intrinsic fluorescence in the wavelength range of 300 nm to 700 nm.

[0034] Intrinsic fluorescence is calculated as irradiation light intensity × 10 -6 A weaker intensity is desirable. Because the intrinsic fluorescence is minimal, the fluorescent light does not interfere with the optical analysis of biomolecules. A suitable range for intrinsic fluorescence is, for example, irradiation light intensity × 10⁻⁶. -8 ~10 -6 That is the case.

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

[0036] Alkali-free glass and borosilicate glass are particularly suitable glass materials. Specifically, commercially available glass products with trade names AF32, AF35, AS87, D263, D263T, B270, MEMPAX, Willow, G-Leaf, EN-A1, and BDA-E are suitable.

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

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

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

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

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

[0042] For all of the aforementioned glass compositions, colored oxides, such as Nd2O3, Fe2O3, CoO, NiO, V2O5, MnO2, CuO, and Cr2O3, can be added as appropriate.

[0043] As a clarifying agent, 0 to 2% by weight of As2O3, Sb2O3, SnO2, SO3, Cl, F, and / or CeO2 can be added, with the total amount of each being 100% by weight.

[0044] Furthermore, it may be advantageous if the surface energy of the microchannel walls is adjusted so that the biomaterial to be analyzed is transported along the walls or adheres to the walls, and this is also included in the present invention. The surface energy can be adjusted, for example, by coating.

[0045] In the advantageous glass element, at least the walls of the microchannels are coated with a coating that repels or adheres to the biomolecule of analysis. To optimize the surface energy in particular in at least some areas, the coating advantageously includes polymers, silanes, oligonucleotides, antibodies, proteins, antigens or nitrocellulose, or inorganic coatings, particularly metal particles, particularly gold and / or iron, or polymer-based nanoparticles or microparticles, or combinations thereof.

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

[0047] Coatings can typically have a thickness of a single molecular layer, or a thickness that completely fills the channel, ranging from nanometers to the entire channel. In the simplest cases, the coating is applied by a dip coating process. Vacuum processes such as chemical vapor deposition (CVD) or atomic layer deposition (ALD) are suitable, especially when the channel diameter is small. Selective application of the coating within the channel can be advantageous, for example, by applying a lift-off material to the outside of the channel or by selectively performing sputtering on the surface of the glass element.

[0048] In particular, the advantageous structural features of the microchannel walls increase the surface area of ​​the channel walls, and these features interact particularly favorably with coatings and / or packings. In the case of adhesive coatings, a larger surface area is provided for the biomaterial being analyzed. In the case of transportable coatings, the coating suppresses or prevents and / or contributes to the clogging of the structural features by the biomaterial. In the case of packings, adhesion of the packing in the channel can be ensured or at least improved.

[0049] In an advantageous glass element, the microchannel is filled with a gel. This gel advantageously has pores. Since these pores can have various diameters depending on the choice and / or composition of the gel, they can accommodate and / or transport biomaterials of various sizes.

[0050] Particularly suitable in the spirit of the present invention are polyacrylamide gels and / or agarose gels. Agarose gels typically 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 typically used for the analysis of relatively small proteins.

[0051] Gels, particularly the aforementioned gels, can be used in the spirit of the present invention as carrier gels for electrophoresis processes, also commonly referred to as gel electrophoresis. This allows for the separation of different small molecules, particularly DNA, RNA, and proteins, from each other for subsequent analysis. In this case, the term gel electrophoresis generally includes the transfer and / or movement of the analyte through a gel, and separation is not necessarily required.

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

[0053] In classical electrophoresis, molecules form characteristic band patterns within microchannels. Molecules can be visualized through coloring, and by associating their characteristic colors with other molecules, they can be identified.

[0054] However, in the case of glass elements according to the present invention, particularly thin glass elements, it is also provided that a gel and / or alternative material may be used to transfer the substance to be analyzed from the bottom surface of the glass element to the top surface of the glass element by electrophoresis. Subsequently, analysis is performed on the top surface by, for example, fluorescence spectroscopy or DNA sequencing using nanopores. These are standard methods and will not be described in detail herein.

[0055] Glass elements in which at least one DNA or RNA-sensitive dye, particularly multiple dyes, are present at the channel exit region or the bottom of the blind hole are especially advantageous.

[0056] When DNA and / or RNA derived from the biomolecule being analyzed are transported through a microchannel, they reach the channel exit and / or the bottom of the blind hole if the microchannel is formed as a blind hole. There, the DNA or RNA can react with a DNA or RNA-sensitive dye applied there. The presence of the corresponding DNA or RNA strand is then detected by characteristic fluorescence, particularly its wavelength.

[0057] Possible suitable dyes include, 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, molecules for polymerase chain reaction, particularly DNA, primers, nucleotides, and / or the enzyme DNA polymerase, are present in the channel exit region or at the bottom of the blind hole. This embodiment is particularly suitable for single-molecule analysis in which the substance to be analyzed is first amplified within the microchannel and / or blind hole, as described herein. In particular, it is evident that when introducing the substance to be analyzed into the microchannel and / or blind hole, it is also possible to introduce the aforementioned molecules into the microchannel and / or blind hole.

[0059] This embodiment offers the special advantage that amplification of cells and / or DNA and / or RNA in the substance to be analyzed is performed in a very small volume within a microchannel (including blind holes). As a result, the time required for the necessary amplification can be reduced. Furthermore, analysis (including detection) of the substance to be analyzed can be performed in this small volume, resulting in good evaluability, and in particular, good signal quality. In particular, the improved accuracy makes it possible to identify rare populations and distinguish subtle differences between cells. In this way, new biomarkers and signature molecules can be identified. Moreover, by analyzing individual molecules and cells in a sample, it is possible to make statements about the heterogeneity of molecules and cells in the sample. In particular, it is possible to visualize individual differences between patients in order to develop personalized therapies based on the specific properties of cells and molecules. Overall, this provides an efficient and reliable analysis with potential applications in personalized medicine.

[0060] The aforementioned glass element is, so to speak, a precursor to the system according to the present invention, and can be used to perform spatial diagnostic methods and / or single-molecule or single-cell diagnostic methods. As described above, in subsequent processes, biomaterials can be analyzed and / or amplified on the glass element or, for example, by DNA sequencing. In this way, the expression patterns of genes and / or proteins and the distribution patterns of DNA markers in individual cells can be determined simultaneously, while positional information within cell complexes or tissue samples is preserved.

[0061] An analysis system for biomaterials, particularly human, animal, or plant cells and / or cellular components, particularly proteins and / or DNA and / or RNA, comprises a glass element having multiple microchannels, in particular the glass element described above, which, in operation, serves to contain and / or transport biomaterials in or through the microchannels from the lower surface to the upper surface of the glass element, and comprises an evaluation unit capable of analyzing the biomaterials in operation. In the spirit of the present invention, the evaluation unit is assigned to or attached to the glass element.

[0062] In general, the evaluation unit detects the presence of biomolecules and / or their components that are transported through and / or present within microchannels. In particular, the evaluation unit may be equipped with an electronic image sensor capable of capturing the intrinsic fluorescence of the aforementioned dyes. Thus, this also includes configuring the microchannels as blind holes.

[0063] Therefore, an advantageous system is provided in which at least one DNA or RNA-sensitive dye is present in the channel exit (22) region and / or blind hole (25), particularly at its bottom, and the color information of the dye can be grasped, particularly spatially, by the evaluation unit.

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

[0065] Advantageously, the system, in its operating state, includes a sample support on which the biological material to be analyzed is placed. This is typically a sample plate or slide.

[0066] A particular advantage is that a transport unit is assigned to the glass element, and this transport unit introduces biological material into the microchannels of the glass element during operation.

[0067] A transport unit is generally a unit that applies a force to the biomaterial being analyzed that at least assists the movement and / or transport of a glass element into and out of the microchannel and toward the opposite surface.

[0068] An advantage is a system in which the transport unit includes a negative pressure unit that applies negative pressure to the outlet side of the glass element in the operating state and / or a unit that generates a potential gradient across the thickness of the glass element to introduce the biomaterial to be analyzed into the microchannel by ion flow or electric field and / or transport it through the microchannel.

[0069] DNA is fundamentally negatively charged due to its phosphate residues, and therefore migrates to the anode during gel electrophoresis. Thus, it is particularly advantageous to have the anode on the upper surface of a favorable glass element, or to have the upper surface designated as the anode. If the cathode is located on or near the lower surface, applying a voltage generates an electric field, causing anions to move to the anode and cations to move to the cathode.

[0070] During DNA analysis, it is naturally possible to amplify DNA fragments beforehand and / or within the glass element channel, particularly by polymerase chain reaction.

[0071] Depending on the biomolecule to be analyzed, it is naturally possible and included in the present invention to place the anode on the lower surface of the glass element, the cathode on the upper surface, and / or assign them to each other. Other arrangements are also possible and included in the present invention. For example, it may be sufficient to move the medium in which the biomolecule to be analyzed is contained by a transport unit.

[0072] Similarly advantageous, the transport unit is equipped with a blade unit that can introduce the biomaterial to be analyzed into microchannels, particularly blind holes, by spreading it.

[0073] An advantageous system is one that incorporates a microfluidic system as a transport unit, which introduces the biomaterial to be analyzed into blind holes and / or microchannels via fluid flow.

[0074] Particularly advantageous, the system can take the form of an advanced system in which the evaluation unit is equipped with a closed membrane or nanoporous membrane on the exit side of the microchannel, which is composed of a non-conductive or semiconductor, or a combination of non-conductive, conductive and / or semi-conductive components, particularly glass, silicon, graphene and / or biomolecules, particularly lipid molecules, DNA origami structures or transmembrane proteins, and this membrane allows for the analysis of biomaterials in particular for their arrangement, binding behavior, size or optical properties, particularly fluorescence.

[0075] In an alternative configuration, the system features an evaluation unit on the top surface of the microchannel in the form of a closed membrane or nanoporous membrane, particularly composed of nonconductors or semiconductors, or combinations of nonconductor, conductive, and / or semiconducting components, especially glass, silicon, graphene, and / or biomolecules, especially lipid molecules or transmembrane proteins. This allows the biomaterial under analysis to be analyzed in terms of its arrangement, binding behavior, size, or optical properties, particularly fluorescence.

[0076] As mentioned above, microchannels can be filled with gel, especially to assist with transport. The gel itself can contain dyes that bind to the biomolecules being analyzed. For DNA analysis, DNA-binding dyes such as ethidium bromide are used. Alternatively, the sample can be in a sample buffer, and this buffer can contain a dye such as crystal violet. It is also possible to color the gel after electrophoresis with DNA-binding dyes such as methylene blue, steinsol, or ethidium bromide. When using fluorescent dyes such as ethidium bromide, it is often necessary to irradiate the agarose gel with UV light and equip the imaging camera with a UV filter. RNA analysis is also possible, of course.

[0077] In advantageous systems, the amplification of biomaterials within microchannels and / or blind holes is provided in the operating state, which is often referred to as digital PCR. In particular, molecules for polymerase chain reaction are present at the channel exit region or the bottom of the blind hole.

[0078] If a blind hole contains a molecule for polymerase chain reaction (PCR), it can amplify DNA or RNA molecules contained in the substance being analyzed. This includes, for example, DNA, primers, nucleotides, and the enzyme DNA polymerase.

[0079] The temperature stability of the glass element allows for standard PCR at temperatures up to approximately 100°C, in addition to the usual room temperature PCR performed at up to 40°C. At these temperatures, unlike plastic, there is no adverse effect on spatial information due to thermal expansion or displacement of blind holes. Furthermore, because glass has better thermal conductivity than plastic, heat is uniformly distributed within the glass element, ensuring homogeneous PCR throughout the sample. In addition, glass has the advantage of having less intrinsic fluorescence than plastic, resulting in high signal intensity against the background.

[0080] As described above, the exit matrix of the microchannels on the upper surface of the glass element can be mapped to the inlet matrix of the microchannels on the lower surface of the glass element. Therefore, in the aforementioned system, the inlet position of the corresponding biomaterial on the lower surface can be determined from the position of the analysis result on the upper surface. In other words, the position of the source of the biomaterial in the sample can be mapped to the position of the microchannels in the glass element. In this way, the glass element and / or system according to the present invention can enable spatial analysis of a biomaterial sample. This is also called "spatial biology" or "spatial diagnostics".

[0081] The aforementioned glass elements can be manufactured efficiently and in large quantities using the method according to the present invention, which is described below. - A method and process for providing an ultrashort pulse laser, - A method and process for providing a glass element substrate, - A process of irradiating a glass element substrate with a pulsed laser beam of an ultrashort pulse laser, in which, - 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 into a focal region that extends long in the beam direction, and this focal region is located at least partially within the glass element substrate. - A method and step in which the intensity and focal range of the laser beam are set to a level such that the laser beam leaves filamentous damage in the workpiece, and multiple such filamentous damages are introduced at that time, - A method and process for forming microchannels by etching filamentous damage. Includes.

[0082] The glass element substrate can generally be a glass plate of desired dimensions. A pulsed laser beam from an ultrashort pulse laser is irradiated onto the glass element substrate, and 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.

[0083] The laser beam is focused into a focal region that extends long in the beam direction, and this focal region is at least partially located within the glass element substrate. For this purpose, various optical configurations can be used, such as a Bessel optical system using axicon, an optical system utilizing chromatic aberration, or an optical system utilizing spherical aberration.

[0084] The laser beam intensity and focal range are set to a level that leaves filamentous damage in the workpiece, and that multiple such filamentous damages are introduced at that time. These filamentous damages represent, so to speak, thread-like damage in glass, and become microchannels when etched in a subsequent process.

[0085] Filamentous damage is caused by nonlinear optical effects involving the self-focusing of the laser beam. This requires ultrashort laser pulses in the ps region.

[0086] A favorable approach is provided in which a series of laser pulses are used to generate filamentous damage.

[0087] In this so-called burst mode, the laser energy is not emitted as a single pulse, but as a series of short, consecutive pulses, which together form a pulse packet, or so-called burst. Such pulse packets typically have slightly higher energy than a single pulse in normal single-shot mode. However, the pulses in a burst themselves have significantly less energy than a single pulse. With respect to pulses within a burst, it is possible to provide flexible control over the pulse energy, in particular, that the pulse energy is substantially constant, or that the pulse energy increases or decreases.

[0088] A suitable laser source can be a neodymium-doped yttrium-aluminum garnet laser with a wavelength of 1064 nanometers. This laser source can generate a raw beam with a diameter of, for example, (1 / e²) 12 mm, and a biconvex lens with a focal length of 16 mm can be used as the optical system. For generating the raw beam, a suitable beam shaping optical system, such as a Galilean telescope, can be used.

[0089] The laser source can operate at repetition rates of 1 kHz to 1000 kHz, preferably 2 kHz to 100 kHz, and especially preferably 3 kHz to 200 kHz.

[0090] Here, the repetition rate and / or scanning speed can be selected so that a desired distance between adjacent filamentous damages is achieved.

[0091] The appropriate pulse duration for a laser pulse can be in the range of less than 100 picoseconds, and preferably less than 20 picoseconds.

[0092] Here, the typical output of the laser source is particularly preferably in the range of 20 to 300 watts. The laser energy is basically below the ablation energy of the substrate. According to one advantageous evolution of the present invention, pulse energies exceeding 400 microjoules are used in bursts to obtain filamentous damage, and more preferably total burst energies exceeding 500 microjoules are used.

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

[0094] An advantage is a method in which the elongated focal region includes the region with the highest energy and the subsequent enlarged portion, and the elongated focal region is positioned such that the enlarged portion is located on the surface of the glass element substrate.

[0095] This method allows for the particularly advantageous formation of tapered microchannels. In essence, filamentous pre-damage is generated by the damaged region, and this region subsequently becomes an enlarged channel entrance during the subsequent etching process.

[0096] Particularly advantageous methods include those in which filamentous damage does not reach at least one surface of the glass element substrate, and openings to this surface are created by a subsequent etching process.

[0097] This can also be achieved by selecting the focal point and, consequently, the intensity of the laser beam. Since the generation of filamentous damage is a nonlinear effect, a localized beam intensity is required to cause damage to the glass overall. In other words, below this critical threshold, there is virtually no damage to the glass, or at least not enough damage to form a filament. Only through subsequent etching does the filamentous damage expand, creating a kind of breakthrough that penetrates the surface of the glass element substrate. The inventors have found that this allows for the fabrication of the channel exit matrix of the microchannel with particular precision. "Particular precision" here means that there is little variation in the diameter of the channel exit, and / or the deviation from the ideal circular diameter of the channel exit is at most slight.

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

[0099] The location of the enlarged channel opening is primarily determined by the focal point. As mentioned above, damage to the glass material expands during the subsequent etching process. The larger the damaged area, the greater the expansion.

[0100] The elongated focal shape is used to adjust the expansion characteristics of the channel entrance. Specifically, it is configured so that the expansion region follows the region with the highest energy when viewed along the beam axis. The elongated focal region is positioned so that the expansion portion lies within the surface area of ​​the glass element substrate.

[0101] During subsequent etching, this becomes an enlarged channel entrance, particularly funnel-shaped. The etching process is advantageously carried out such that the microchannel has a diameter of at least 5 μm to 200 μm, preferably 7 μm to 130 μm, and particularly preferably 10 μm to 100 μm in the channel entrance region.

[0102] In one modified form of this method, filamentous damage does not reach at least one surface of the glass element substrate, and openings to this surface are provided by a subsequent etching step.

[0103] This is particularly important because it provides the most accurate geometry possible for the channel exit, which may be located within the cylindrical region of the microchannel. To adjust the channel exit geometry, it has been found advantageous to prevent filamentous damage from penetrating the surface of the glass element where the channel exit is located. In this case, the filamentous damage terminates, so to speak, below this surface. This is also adjusted by the corresponding focal position. In this case, the subsequent etching process forms the opening of the microchannel, i.e., the channel exit.

[0104] The surface of the glass element substrate can be treated by grinding and polishing, particularly before and / or after etching. Grinding and polishing after the etching process has proven particularly advantageous because it can achieve a particularly flat surface, even around the channel inlet and / or channel outlet.

[0105] A favorable method involves introducing a recess, which is a hemispherical, dome-shaped indentation, into the inner wall of the channel by etching.

[0106] This is particularly useful when a slow etching process is being carried out, where the etching rate is less than 15 μm per hour.

[0107] The dome-shaped recess is presumed to be due to a structural component that occurs when filamentous damage is introduced. A liquid etching solution is particularly advantageous as the etching medium. According to this embodiment, etching is performed wet chemically. This can also be advantageous in removing glass components from the surface during etching.

[0108] Both acidic and alkaline solutions can be used as etching solutions. Suitable acidic etching media include HF, HCl, H2SO4, ammonium difluoride, HNO3 solutions, or mixtures of these acids. Suitable basic etching media include KOH or NaOH alkaline solutions. Acidic etching solutions can usually achieve higher removal rates. However, basic solutions are preferred, especially when only slow removal is desired.

[0109] Etching is preferably carried out in a temperature range of 40°C to 150°C, and particularly preferably 50°C to 120°C. Generally, silicate glass with a low alkali content is particularly suitable for structuring according to the present invention. If the alkali content is too high, etching becomes difficult. Therefore, according to one advanced form of the present invention, the glass of the glass element is provided to be silicate glass having an alkali oxide content of less than 17% by weight.

[0110] The aforementioned glass elements equipped with microchannels can be advantageously used for the analysis of biomaterials, particularly human, animal, or plant cells and / or cellular components, especially proteins and / or DNA.

[0111] As an alternative to spatial diagnostics, glass elements, with their numerous blind holes, can be used for single-molecule or single-cell diagnostics. In this case, liquid samples, in particular, are introduced into the blind holes by spreading them, for example, using a blade process or the process described above. Depending on the concentration chosen, this results in only a small amount, or just one, of the target molecule or cell entering each blind hole. Subsequently, the target molecule and / or cell can be increased as desired within each blind hole for analysis, especially fluorescence analysis.

[0112] In particularly advantageous embodiments, it is provided that individual blind holes or groups of blind holes can be controlled as desired, and that specific tests be performed on each of these blind holes or groups of blind holes. This is particularly advantageous for determining the heterogeneity of samples and for medical diagnostic methods (highplex diagnostics) that allow for the performance of numerous different tests in a small space.

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

[0114] [Figure 1] Figures 1a to 1d are cross-sectional views of various embodiments of a glass element with an enlarged microchannel. [Figure 2] This is a cross-sectional view showing a portion of a glass element with enlarged microchannels. [Figure 3] This is a diagram showing an electron microscope image of the wall of a microchannel. [Figure 4] Figures 4a and 4b are top views of the top and bottom surfaces of the glass element. [Figure 5] This figure shows a microscopic image of a cross-section of a glass element. [Figure 6] This is a diagram showing a spatial analysis system for biomaterials. [Figure 7] This diagram shows the layout of the dye on the upper surface of the glass element. [Figure 8] This diagram shows the layout of dyes in blind holes of a glass element. [Figure 9] This figure shows a laser processing apparatus for glass elements in preparation for subsequent etching. [Figure 10] Figures 10a and 10b schematically show the influence of the structural components of the microchannel wall.

[0115] Figure 1a shows one embodiment of a glass element (1) having multiple microchannels (2). In this embodiment, the microchannels connect the upper surface (O) and the lower surface (U) of the glass element and penetrate the glass element. As mentioned above, the upper surface generally refers to the surface or face of the glass element that is opposite to the sample or the side on which the analysis is performed. The lower surface is usually the surface or face of the glass element that faces the sample, i.e., the biological material being analyzed. In the terminology used herein, the channel inlet (21) is located in the region of the lower surface (U), and the channel outlet (22) is located in the region of the upper surface (O).

[0116] In this example of Figure 1a, the microchannel tapers from the bottom surface (U) to the top surface (O), and this taper is continuous in this example. Generally, the widening in at least some areas is referred to as funnel-shaped in this specification. In the illustrated example, the taper is continuous. The profile of the microchannel (2) can also be referred to here as a frustocone. The tapered region extends here over the entire thickness (s) of the glass element.

[0117] The thickness of the processed glass element or material thickness (s) can be 0.1 mm to 3 mm, as described above. The illustrated glass element (1) has substantially parallel surfaces (O) and (U). Usually, the same is true for the sides. The glass element (1) is advantageously slide-shaped.

[0118] Figure 1b shows a similar embodiment in which the tapering of the microchannel is more pronounced. Because the microchannel (2) tapers toward the upper surface (O) and / or is funnel-shaped in at least some areas, in the spirit of the present invention, the diameter (dU) of the channel inlet (21) on the lower surface (U) is larger than the diameter (dO) of the channel outlet (22) on the upper surface (O) of the glass element (1).

[0119] Figure 1c shows a further embodiment in which the microchannel profile consists of a tapered frustoconical region in the lower (U) region and a cylindrical region in the upper (O) region. The overall shape can similarly be described as funnel-shaped. As described above, the pitch (P) is determined by the center point of the channel outlet (22). Since a smaller pitch results in a higher microchannel density, the smallest possible pitch is aimed for. The pitch can be 3 to 70 μm, as previously mentioned. 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 passing perpendicularly through the surfaces (O) and (U) of the glass element.

[0120] Figure 1d shows an embodiment in which the microchannel (2) tapers towards the bulk of the glass material from both the bottom (U) and top (O) surfaces. In the shape shown here as an example, this is hourglass-shaped or X-shaped. This is, in principle, like two inverted frustums stacked on top of each other with their short sides in contact.

[0121] Figure 2 shows a portion of the glass element (1) according to Figure 1b. The aperture angle α is indicated, which should be interpreted as the angle between the channel walls in the enlarged region of the microchannel (2). Advantageously, the aperture angle α is between 0.1° and 30°, and especially advantageously between 2° and 18°, and it should be understood that the upper and lower limits of this interval are, of course, variable.

[0122] As explained in detail above, this tapered microchannel makes it possible to achieve two conflicting objectives: on the one hand, to aim for the smallest possible pitch to increase the resolution of the analysis, thereby achieving high detection sensitivity and ultimately a larger channel entrance diameter; and on the other hand, to ensure the mechanical stability of the glass element in order to use the glass element efficiently. Mechanical stability is maintained, especially in the non-tapered region, where a large amount of material remains.

[0123] Figure 3 shows an electron microscope image of the wall of a microchannel (2) of a glass element (1) manufactured by the inventors according to the present invention, and in particular shows a portion of the microchannel region (40) as shown in Figure 1c. As already stated herein, the lateral dimension or diameter of the channel (2) is less than 200 micrometers, and in this example it is about 14 micrometers. A diameter in the range of 5 micrometers to 200 micrometers is preferred, but is not limited to the example described herein.

[0124] Dome-shaped or rounded cap-shaped recesses (7) resulting from etching of filamentous damage introduced into the glass element by an ultrashort pulse laser beam are clearly visible. The depth of the dome-shaped recesses (7) is typically less than 5 μm, and the lateral dimension is typically 5 to 20 μm.

[0125] As a result of the etching process of the filamentous damage, the dome-shaped recesses (7) are adjacent to each other, and the concave curves of the recesses (7) are tangent to each other, forming ridges (70). Furthermore, when viewed from above the recesses (7), it can be seen that the ridges (70) form the polygonal boundary lines (71) of the recesses (7). Here, the average number of corners (72) of the boundary lines (71) of the recesses (7) is favorably less than 8, and favorably less than 7. The latter characteristic occurs when the area occupied by the majority of the dome-shaped recesses is mathematically convex.

[0126] The ridge (70) of channel 2 shown in Figure 3 is very narrow, and there is no recognizable region where each concave curve of the recess (7) merges with one convex curved region on the ridge (70). Therefore, according to one advanced form of the present invention, the structure of channel (2) can also be described as having a convex region area ratio of less than 5%, preferably less than 2%, in channel (2).

[0127] Figure 4a schematically shows top views of the top (O) and bottom (U) surfaces of the glass element (1). The diameter of the channel inlet (21) and the diameter of the channel outlet (22) are substantially equal. Such a pattern can be assumed, for example, in the embodiment shown in Figure 1d.

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

[0129] As shown in Figures 4a and 4b, the channel inlet and channel outlet form a matrix. Since the position of the channel inlet corresponds to the position of the channel outlet, if the position of the channel outlet is known, spatial measurements can be performed on the channel inlet side, and therefore on the sample side, for the presence of a certain event, such as DNA or a specific biomolecule.

[0130] Figure 5 shows a photograph taken with an optical microscope of a cross-section of a glass element (1) manufactured by the inventors, which substantially corresponds to the embodiment shown in Figure 1c. A cylindrical region of the microchannel (2) follows a funnel-shaped magnified region. 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 embodiment, SCHOTT glass D263 was used. The present invention is not limited to any particular type of glass. In other embodiments, borosilicate glass and / or alkali-free glass were manufactured. Borosilicate glass, in particular, is frequently used in laboratories due to its chemical resistance. As already mentioned, glass that exhibits only slight intrinsic fluorescence in the visible region of the spectrum is particularly advantageous.

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

[0132] A gel, particularly the aforementioned gel, can be present within the microchannel (2). This is not shown in Figure 6. On the upper surface of the glass element (1), at least one DNA or RNA-sensitive dye (610) is applied to the channel exit (22) region. When a biomaterial (60) passes through the microchannel (2), particularly using the gel, it comes into contact with the dye (610) on the upper surface. The typical color of the dye can indicate its presence, and in some cases, the presence of a specific DNA strand can be indicated. As previously mentioned, the dye is usually irradiated with excitation light, and then the typical fluorescence of the dye is measured using an evaluation unit. For clarity, the evaluation unit is not shown in this figure. Similarly, as previously mentioned, this can be a digital camera.

[0133] Advantageously, different dyes (610) may be applied to different channel outlets (22). In that case, different DNA or DNA strands can be identified when analyzing biomaterials.

[0134] The top view shows a pattern in the illustrated system (10) that allows for the spatial estimation of the presence of specific biomolecules, particularly DNA. Everything described for DNA also applies to RNA.

[0135] The transport of biomaterial (60) into or through the microchannel (2) can be assisted by the presence of a transport unit (65). In the example in Figure 6, this is a unit that generally utilizes electric potential for the transport of biomaterial. Illustrated is a capacitor unit in which a voltage source (V) is connected to electrodes (651, 652). In this example, the electrodes are located on the underside of the sample support (62) and on a non-conductive plate (653), particularly a glass plate or plastic. Thus, the electrodes form a plate capacitor, with the biomaterial (60) and glass element (1) between the plates. When a voltage is applied, an electric field is generated, and this electric field pulls or pushes the biomaterial and / or the surrounding medium toward the electrode (651).

[0136] When the non-conductive plate (653) is brought into contact with the upper surface (O) of the glass element, the non-conductive plate (653) can also function as a closure for the microchannel (2). In this case, a blind hole is formed, so to speak, and the dye is not removed from the surface of the glass element by the progressing biological material.

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

[0138] Other configurations are also possible and are included in the present invention. In particular, it is also possible to transport biological materials using pressure and / or negative pressure instead of electromotive force.

[0139] Figure 7 shows a top view of a glass element (1) with sensitive dyes (610, 611, 612, 613, 614) on its top surface (O), particularly in the system shown in Figure 6. The illustrated pattern schematically represents possible analytical results.

[0140] Figure 8 shows, for clarity, an embodiment of a glass element having a microchannel (2) formed as a blind hole (25). The aforementioned dyes (610, 611) are applied to the bottom of the blind hole. The electrodes (651) of the electrical transport unit (65) may simply be applied to the surface of the glass element. In this case, the electrodes are particularly transparent electrodes, which can be manufactured, for example, as an ITO coating, in order to detect the dye fluorescence through the glass material located above the bottom of the blind hole. Alternatively, the electrodes may be formed as a raster and / or grid, particularly having a void above the bottom of the corresponding blind hole (25).

[0141] Figure 9 shows an embodiment of the laser processing apparatus (20), in which filamentous damage (32) can be introduced into a glass element substrate (1), and then microchannels can be introduced at the location of the filamentous damage (32) by an etching process. The apparatus (20) comprises an ultrashort pulse laser (300) to which a focusing optical system (230) is pre-connected, and a positioning unit (170). The positioning unit (170) can laterally position the point (73) of the collision of the laser beam (270) of the ultrashort pulse laser (300) with the surface (O) of the plate-shaped glass element (1) to be processed. In the illustrated example, the positioning unit (170) is equipped with an XY table, on which the lower surface (U) of the glass element (1) is placed. However, alternatively or additionally, the optical system can be made movable to move the laser beam (270), thereby allowing the collision point (73) of the laser beam (270) to be moved while the glass element (1) remains fixed.

[0142] Figures 10a and 10b schematically illustrate the effect of the structure of the microchannel wall. Figure 10a shows a smooth channel wall, while Figure 10b shows a structured channel wall with roughness Ra. The channel wall has an opening angle. The structure can have a dome-shaped recess. EOF represents electroosmotic flow. As mentioned above, electroosmotic flow is the charge-induced movement of a liquid along a polar surface in an electric field. As shown in Figure 10b, if the roughness of the polar surface is large, the EOF velocity decreases compared to a smooth surface as shown in Figure 10a, and the direction of movement of polar biomaterials within the glass element may be reversed.

[0143] The focusing optical system (230) focuses the laser beam (270) to a focal point that is elongated in the beam direction, i.e., a focal point that is elongated laterally, and especially perpendicularly, to the irradiation surface (O). Such a focal point can be generated, for example, using a conical lens (so-called axicon) or a lens having large spherical aberration and / or large chromatic aberration. The positioning unit (170) and the ultrashort pulse laser (300) are controlled, advantageously by a programmed computer unit (150). In this way, a predetermined pattern of filamentous damage (32) distributed laterally along the surface (O) can be generated, which is particularly advantageous by reading position data from a file or via a network. The shape of the filamentous damage (32) can be adjusted by the position of the focal point, and this adjustment is made such that a localized expansion of 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 and focused by 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 about 10 ps. The pulses are emitted in bursts of two or more pulses, preferably four or more pulses. The burst frequency is 12 to 48 ns, advantageously about 20 ns, the pulse energy is at least 200 microjoules, and accordingly the burst energy is at least 400 microjoules.

[0145] Subsequently, after introducing one or more filamentous damages (32), the glass element (1) is removed and stored in an etching bath, in which the glass is removed along the filamentous damages (32) by an etching process in the bath, thereby introducing one microchannel (2) at each location of such damage (32) into the glass element 1.

[0146] A basic etching bath with a pH value >12 is preferred, for example, a KOH solution of >4 mol / l, preferably >5 mol / l, particularly preferably >6 mol / l, but <30 mol / l is preferred. According to one embodiment of the present invention, 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 present invention have the advantage of enabling spatial analysis of biomaterials. The glass elements can be manufactured efficiently and are composed of materials commonly used in laboratory environments. After use, the glass can be melted again, and thus the raw materials can be recycled.

Claims

1. A system (10) for measuring a biological substance (60), A sample support (62), wherein the biological substance (60) is present on the sample support (62), and the sample support (62) and The glass element (1) in contact with the aforementioned biological material (60), A pair of electrodes (651, 652), wherein the glass element (1) is positioned between the pair of electrodes (651, 652), Equipped with, The glass element (1) comprises a plurality of microchannels (2), the microchannels (2) connecting one surface (U, O) of the glass element (1) to the opposite surface (O, U) of the glass element (1), or terminating within a blind hole (25). The microchannel (2) tapers from the bottom surface (U) to the top surface (O). Multiple different DNA or RNA-sensitive dyes (610, 611, 612, 613, 614) are present in the region of the multiple channel exits (22) or at the bottom of the multiple blind holes (25) in the upper surface (O) region. Measurement system (10), wherein when a voltage is applied to the pair of electrodes (651, 652), an electric field is generated, and this electric field pulls or pushes the biomaterial (60) and / or the surrounding medium toward one of the electrodes (651), thereby the glass element (1) plays a role in containing and / or transferring the biomaterial (60) in or through the microchannel (2) from the lower surface (U) of the glass element to the upper surface (O) of the glass element.

2. The measurement system (10) according to claim 1, wherein the diameter (dU) of the channel inlet (21) of the lower surface (U) microchannel is greater than the diameter (dO) of the channel outlet (22) or channel end (25).

3. The measurement system (10) according to claim 1 or 2, wherein the microchannel (2) has a diameter (dU, dO) of 5 μm to 200 μm in at least the channel inlet (21), channel outlet (22), or channel end (25) region of the glass element (1).

4. The microchannel (2) is formed in a funnel shape in at least a portion of its region. The measurement system (10) according to claim 2, wherein a cylindrical or funnel-shaped region is adjacent to the funnel-shaped region of the channel inlet (21).

5. The density of the microchannels is at least 20 / mm². ２ The measurement system (10) according to claim 1 or 2.

6. The measurement system (10) according to claim 1 or 2, wherein the channel wall of the microchannel (2) has a structure comprising a plurality of rounded, dome-shaped recesses (7).

7. The measurement system (10) according to claim 2, wherein molecules for polymerase chain reaction are present in the region of the channel outlet (22) or at the bottom of the blind hole (25).

8. The measurement system (10) according to claim 1, further comprising an evaluation unit capable of analyzing the biological substance in an operating state.

9. The measurement system (10) according to claim 8, wherein the color information of the DNA or RNA-sensitive dye can be obtained by the evaluation unit.

10. The measurement system (10) according to claim 8 or 9, wherein, in operation, amplified a biological substance within a microchannel (2) and / or a blind hole (25).

11. The measurement system (10) according to claim 8 or 9, wherein the evaluation unit is provided with a closure membrane or a nanoporous membrane on the outlet side of the microchannel (2).

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