Abstract
The invention relates to an electrophoresis device (1) for use in a method for producing transparent biological samples (2), comprising a reaction frame (3), the reaction frame (3) having an open top side (4) and a bottom side (5) opposite the top side (4), characterized in that the bottom side (5) has at least partially one opening (6). The invention also relates to a use of a sample cassette (19) for an electrophoresis method.
Claims
1. An electrophoresis device for use in a method for producing transparent biological samples, comprising a reaction frame, wherein the reaction frame has an open top side and a bottom side opposite the top side, wherein the bottom side at least partially comprises an opening.
2. The electrophoresis device as claimed in claim 1, wherein the reaction frame can be inserted into a receiving vessel, wherein the receiving vessel has a base plate, and wherein the reaction frame realizes, with the receiving vessel, a reaction chamber.
3. The electrophoresis device as claimed in claim 1, wherein the reaction frame has a cover plate, which is fixedly or detachably connected to the top side of the reaction frame, wherein the cover plate substantially completely covers the top side of the reaction frame.
4. The electrophoresis device as claimed in claim 1, wherein the electrophoresis device has a first electrode and a second electrode.
5. The electrophoresis device as claimed in claim 1, wherein the reaction frame comprises four inner side walls, which are arranged in a cuboid shape in relation to one another.
6. The electrophoresis device as claimed in claim 4, wherein the receiving vessel has four outer side walls, which are arrange in a cuboid shape in relation to one another, wherein the four outer side walls have an inner circumference that is larger than an outer circumference of the inner side walls.
7. The electrophoresis device as claimed in claim 1, wherein the reaction frame comprises an inner hollow cylinder.
8. The electrophoresis device as claimed in claim 7, wherein the receiving vessel comprises an outer hollow cylinder, wherein the outer hollow cylinder has an inner circumference that is larger than an outer circumference of the inner hollow cylinder.
9. The electrophoresis device as claimed in claim 5, wherein the first electrode is arranged, on the reaction chamber side, on one of the inner side walls, and that the second electrode is arranged, on the reaction chamber side, on the inner side wall that is opposite the first electrode.
10. The electrophoresis device as claimed in claim 7, wherein that the first electrode is arranged, on the reaction chamber side, on the inner hollow cylinder.
11. The electrophoresis device as claimed in claim 1, wherein the electrophoresis device has a sample cassette.
12. The electrophoresis device as claimed in claim 11, wherein the electrophoresis device comprises a cassette holder, into which the sample cassette can be clamped.
13. The electrophoresis device as claimed in claim 11, wherein the sample cassette comprises a base element and a cover element, which can be plugged together to form a cassette enclosing the sample, wherein the base element and the cover element each have a multiplicity of perforations arranged in a grid-like manner.
14. The electrophoresis device as claimed in claim 11, wherein the reaction frame has a receiving profile for receiving the sample cassette or for receiving the cassette holder.
15. The electrophoresis device as claimed in claim 11, wherein the reaction chamber is divided into a first reaction compartment and a second reaction compartment when the sample cassette or the cassette holder is received in the receiving profile.
16. A use of a sample cassette as claimed in claim 11 for an electrophoresis method.
17. The use of a sample cassette as claimed in claim 16, wherein the electrophoresis method is an electrophoresis method for producing transparent biological samples.
Description
BRIEF DESCRIPTION OF THE FIGURES
[0033] Further features, details and advantages of the invention are given by the wording of the claims and by the following description of exemplary embodiments, with reference to the drawings.
[0034] FIG. 1 shows an electrophoresis device according to a horizontal embodiment, in a schematic plan view;
[0035] FIG. 2 shows an electrophoresis device according to a horizontal embodiment with a sample cassette and a receiving vessel, in a schematic plan view;
[0036] FIG. 3 shows an electrophoresis device according to a horizontal embodiment with a receiving vessel and a cover plate, in a longitudinal vertical cross-section;
[0037] FIG. 4 shows an electrophoresis device according to a horizontal embodiment with a receiving vessel and a cover plate, in a transverse vertical cross-section;
[0038] FIG. 5 shows an electrophoresis device according to a vertical embodiment, in a vertical cross-section;
[0039] FIG. 6 shows an electrophoresis device according to a further embodiment with a gap, in a vertical cross-section;
[0040] FIG. 7 shows an electrophoresis device according to a further vertical embodiment, in a vertical cross-section;
[0041] FIG. 8 shows an electrophoresis device according to a further vertical embodiment, in a vertical cross-section;
[0042] FIG. 9 shows a preferred embodiment of a sample cassette.
DETAILED DESCRIPTION OF THE FIGURES
[0043] FIG. 1 shows an electrophoresis device 1 according to a horizontal embodiment, in a schematic plan view. The electrophoresis device 1 shown, which is intended for use in a method for producing transparent biological samples 2, preferably comprises a sample cassette (not shown) 19 as well as a reaction frame 3. The reaction frame 3 is made up of four inner side walls 13a-d, with the inner side walls 13a-d being connected to one another in such a manner that they form a cuboid that is open on two sides, and thus a reaction chamber 9 that is open on two sides. The reaction frame 3 is designed so as to be open on a top side 4 (FIG. 3) and on a bottom side 5 (FIG. 3). In this case the top side 4 of the reaction frame 3 is arranged above a horizontal central axis B (FIG. 3) of the reaction frame 3, whereas the bottom side 5 is arranged beneath the horizontal central axis B. In the case of the reaction frame 3 as shown in FIG. 1, the sample preparation 2 (not shown) and the electrodes 11, 12 are arranged substantially in the same horizontal plane. This allows the reaction liquid 27 to be changed easily. In particular, there is no need for the old reaction liquid 27 to be tipped out of the reaction frame 3. At the same time, the strength of the electric current can be influenced, for example by changing the buffer solution 27.
[0044] The reaction frame 3 shown in FIG. 1 has, in the reaction chamber 9, a first electrode 11 with a first electric leadthrough 36 for contacting to the power source, and a second electrode 12 with a second electric leadthrough 37 for contacting to the power source. The first and the second electrode 11, 12 can be connected to a DC voltage source via the electric leadthroughs 36, 37 in order to generate an electric field.
[0045] FIG. 1 additionally shows a receiving profile 24 in the form of grooves 33. The receiving profile 24 serves to receive the sample cassette 19 or to receive the cassette holder 20. According to the embodiment shown, the receiving profile 24 comprises grooves 33, which are each realized, on the reaction chamber side, on two opposite inner side walls 13a,c and extend substantially in the vertical direction.
[0046] It may be provided in this case that the grooves 33 taper toward the bottom side 5 of the reaction frame 3, thereby preventing the sample cassette 19, or the cassette holder 20, from sliding downward (i.e. toward the bottom side 5) through the grooves 33 of the reaction frame 3. The locking mechanism thus enables the reaction frame 3 to be transferred quickly and safely from one receiving vessel 7 to another without the risk of loss of the sample cassette 19, or of the cassette holder 20. A receptacle profile 24 realized in such a manner thus has the advantage that the sample cassette 19, and the cassette holder 20, can be easily and reliably inserted into the receiver 24.
[0047] FIG. 2 shows an electrophoresis device 1 according to a horizontal embodiment with a sample cassette 19 and a receiving vessel 7, in a schematic plan view. The receiving vessel 7, which in the case of the horizontal embodiment is preferably a tank, has four outer side walls 14a-d which, together with a base plate 8 (FIG. 3), form a cuboid open toward the upper side. The upper side 4 of the frame 3 is understood to be the side that faces away from the base plate 8 of the tank 7. Correspondingly, the bottom side 5 of the reaction frame 3 is the side that faces toward the base plate 8 of the tank 7. The outer side walls 14a-d span an inner circumference 15a that is larger than the outer circumference 16a of the reaction frame 3, which is also cuboid-shaped. As a result, there is a distance between the outer side walls 14a-d of the tank 7 and the inner side walls 13a-d of the reaction frame 3. This distance should be large enough to allow the frame 3 to be inserted into and removed from the tank quickly and easily. Preferably, the distance should be at least 0.5 cm. When the tank 7 is filled with buffer 27, the reaction frame 3 and the bottom plate 8 form a reaction chamber 9 that is open on one side. The reaction frame 3 can then be removed from the receiving vessel 7 at any time and transferred to another receiving vessel 7 filled with fresh reaction liquid 27, without thereby transferring old reaction liquid 27. The reaction liquid 27 can then drain off through the opening 6 in the bottom side of the frame 3. This allows easy changing of the buffer. In particular, the device 1 does not have to be tipped.
[0048] The sample cassette 19 is inserted into the grooves 33 as shown in FIG. 2. In this case, when the sample cassette 19 is received in the grooves 33, the reaction chamber 9 is divided into a first reaction compartment 25 and a second reaction compartment 26. As a result, the biological sample 2 is arranged substantially centrally in the reaction chamber 9, where an approximately homogeneous electric field is concentrated between the electrodes 11, 12. The current flow in this case is from one of the electrodes 11, 12 (anode) to the other (cathode), and passes through the sample 2. In this way the contaminating components are removed from the sample 2, with negatively charged ions migrating to the anode, and positively charged ions to the cathode.
[0049] FIG. 3 shows the electrophoresis device 1 according to a horizontal embodiment with a receiving vessel 7 and a cover plate 10, in a longitudinal vertical cross-section. The sample cassette 19 in this case is clamped in a cassette holder 20, which in turn is received by the grooves 33. In order that the cassette holder 20, in which the sample cassette 19 is enclosed, can divide the reaction chamber 9 into two reaction compartments 25, 26, the cassette holder 20 is realized in such a manner that it protrudes above the surface of the reaction liquid 27 when it is received in the receiving profile 24 of the frame 3. This ensures that the current flowing between the electrodes 11, 12 during the electrophoretic clearing process passes exclusively through the tissue sample 2.
[0050] FIG. 3 further shows how the reaction frame 3 is received in the receiving vessel 7. It can be seen how the inner side walls 13a-d together with the base plate 8 form a reaction chamber 9, which in turn is divided into a first 25 and a second reaction compartment 26 when the cassette holder 20 is received in the grooves 33. In addition, a cover plate 10 is shown.
[0051] FIG. 4 shows an electrophoresis device 1 according to a horizontal embodiment with a receiving vessel 7 and a cover plate 10, in a transverse vertical cross-section.
[0052] According to the design shown, the first electrode 11 and the second electrode 12 are each realized in the form of a rod electrode. However, plate electrodes 11, 12 or electrodes 11, 12 realized as a grid may also be provided. Plate electrodes 11, 12 have the advantage that the electric field is evenly distributed over the entire reaction chamber 9 and is not just limited to a particular region within the reaction liquid 27. It is further advantageous if the first electrode 11 and the second electrode 12 are each in electrical contact with the power source via an electric leadthrough 36, 37. The electric leadthroughs 36, 37 make it particularly easy to connect the electrodes 11, 12 to a voltage device.
[0053] As can also be seen from FIG. 3 and FIG. 4, the device 1 shown has a cover plate 10 that can be attached to the top side 4 of the reaction frame 3. The cover 10 may, for example, be placed or plugged onto the frame 3. The cover 10 completely closes off the reaction frame 9 and the receiving vessel 7 such that no foreign bodies can enter the reaction chamber 9. This ensures a safe and clean clearing method. In particular in this case, it may be provided that at least one of the electric contacts for connecting the electric leadthroughs 36, 37 to a power source is recessed in the cover plate 10, such that the cover plate 10 needs to be attached to, or placed on, the reaction frame 3 in order to connect the device 10 to the power source. In this case, the electric contacts can simultaneously serve as plug connections for attaching the cover plate 10 to the reaction frame 3. This ensures that current can only flow in the reaction chamber 9 when the cover plate 10 closes the reaction frame 3, which serves as an additional safety aspect.
[0054] Additionally shown in FIG. 4 are the cassette holder 20 and the sample cassette 19, the sample cassette 19 being received in the cassette holder 20. For the exemplary embodiment, however, it is also conceivable for the sample cassette 19 to be inserted directly into the grooves 33 without the provision of an additional cassette holder 20. According to the design, it is provided that the cassette holder 20 is slid into the grooves 33 of the receiving profile 24, as a result of which the sample 2 is fixed in the center of the reaction frame 3 and thus in the electric field. The sample cassette 19 has perforations 23 configured in the form of a grid. The perforations 23 allow the buffer 27 to reach the tissue sample 2, such that the electric current removes the desired substances from the sample 2.
[0055] FIG. 5 shows an electrophoresis device 1 according to a vertical embodiment, in a vertical cross-section. In the case of this embodiment, the first electrode 11, the sample 2 and the second electrode 12 are arranged substantially vertically with respect to each other. The electrophoresis device 1 shown comprises a receiving vessel 7 with a base plate 8 and with an outer hollow cylinder 18 realized perpendicularly to the base plate 8, the outer hollow cylinder 18 being rotationally symmetrical about a vertical central axis A. The vertical electrophoresis apparatus 1 further comprises a reaction frame 3, which has an inner hollow cylinder 17 and a cover plate 10, the inner hollow cylinder 17 likewise being rotationally symmetrical about the vertical central axis A. In this case, the outer circumference 16b of the inner hollow cylinder 17 is smaller than the inner circumference 15b of the outer hollow cylinder 18. According to the embodiment shown, it is provided that an annular interspace 31 is realized between the inner hollow cylinder 17 and the outer hollow cylinder 18. This allows the frame 3 to be slid easily and smoothly into the receiving vessel 7.
[0056] A vertical electrophoresis device 1 according to FIG. 5 enables the sample cassette 19, together with the tissue sample 2, to be arranged horizontally, resulting in the reaction chamber 9 being divided into an upper first reaction compartment 25 and a lower second reaction compartment 26 when the cassette holder 20, or the sample cassette 19, is received in the electrophoresis device 1. For the purpose of receiving the cassette holder 20 or the sample cassette 19, the inner hollow cylinder 17 has a receiving profile 24 on its side that faces toward the reaction chamber 9, the receiving profile 24 being realized, according to the embodiment shown in FIG. 5, as an annular support for the sample cassette 19.
[0057] The design shown in FIG. 5 also allows easy buffer exchange: for this purpose, the reaction frame 3, comprising the inner hollow cylinder 17 and the cover plate 10, is removed together with the sample cassette 19 from the outer hollow cylinder 18 of the receiving vessel 7 and inserted into an outer hollow cylinder 18 of a second receiving vessel 7 filled with fresh buffer 27. The contaminated reaction liquid 27 can then drain off through the opened vent hole 28 and the opening 6 in the bottom side 5 of the cylindrical frame 3. To open the vent hole 28, it is necessary only to pull the vertical pin 29 out of the vent hole 28.
[0058] Alternatively, it is possible to fill the two reaction compartments 25, 26 separately, i.e. separately from each other, with reaction liquid 27. For this purpose, the cover plate 10 has a through-hole 30 for filling the upper reaction compartment 25 with reaction liquid 27. The through-hole 30 is centrally recessed in the cover plate 10 and realized in principle so to be closable. However, it should be open during the electrophoresis process so that the gas produced at the electrodes 11, 12 can escape. Preferably, the sample cassette 19 or the cassette holder 20 has a vent hole 28, so that the gas 34 produced at the lower electrode 12 (FIG. 5) can be discharged into the upper reaction compartment 25.
[0059] The first electrode 11 is attached to an end region of the inner hollow cylinder 17 near the top side 4, or the cover plate 10, namely on the side of the inner hollow cylinder 17 that faces toward the reaction chamber 9. According to FIG. 5, the second electrode 12 is attached substantially concentrically in the receiving vessel 7, on a side of the base plate 8 that faces toward the reaction chamber 9. According to the embodiment shown in FIG. 5, the first electrode 11 and the second electrode 12 are each of an annular shape. This ensures that the electric field is evenly distributed over the entire reaction chamber 9.
[0060] FIG. 6 shows an electrophoresis device 1 according to a further vertical embodiment with gap 32, in a vertical cross-section. The gap 32 is results from the inner hollow cylinder 17 being connected to the cover plate 10 and extending vertically toward the base plate 8, and the outer hollow cylinder 18 extending vertically from the base plate toward the cover plate 10, the inner hollow cylinder being of a first height C that is less than a second height D of the outer hollow cylinder 18. The embodiment shown in FIG. 6 differs from the embodiment according to FIG. 5 in that the second electrode 12 is attached, in the receiving vessel 7, on a side of the inner hollow cylinder 17 that faces away from the reaction chamber 9. In particular in this case, it is provided that the second electrode 12 is attached to the end of the inner hollow cylinder 17 on the base plate side, i.e. slightly above the gap 32. In order that the gas bubbles produced at the second electrode 12 during electrophoresis can escape from the electrophoresis device 1, the outer hollow cylinder 18 may have openings or perforations (not shown) in a region close to the cover plate 10, via which the gas bubbles can be released to the external environment. It is also conceivable for the cover plate 10 to rest only partially on the outer hollow cylinder 18, such that a small gap remains between the outer hollow cylinder 18 and the cover plate 10 when the cover plate 10 is attached to the inner hollow cylinder 18. Gas bubbles can then likewise escape through this upper gap. According to the embodiment shown in FIG. 6, the first electrode 11 and the second electrode 12 are each of an annular shape. Both electrical leadthroughs 36, 37 of this embodiment are arranged in the cover plate 10.
[0061] FIG. 7 shows an electrophoresis device 1 according to a further vertical embodiment, in a vertical cross-section. It differs from the electrophoresis device 1 shown in FIG. 6 only in that the second electrode 12 is attached to the inner side of the outer hollow cylinder 18. In this case, the inner side is the side that faces toward the inner hollow cylinder 17. The second electrode is also attached to the end of the outer hollow cylinder 18 on the base plate side, slightly above the gap 32. According to the embodiment shown in FIG. 7, the second electrical leadthrough 37 is arranged in the base plate 8.
[0062] It is additionally provided that the cassette holder 20 has a vent hole 28 (FIG. 7 and FIG. 8) that connects the first reaction compartment 25 to the second reaction compartment 26. For the exemplary embodiment, however, it is also conceivable for the sample cassette 19 to be inserted directly into the receiving profile 24 without provision an additional cassette holder 20. In this case, the sample cassette 19 itself may have a vent hole 28. Furthermore, it is conceivable for the receiving profile 24 to have a vent hole 28. The vent hole 28 serves primarily to remove gas or air bubbles 34 produced during filling of the lower compartment 26 with buffer 27. The vent hole 28 then allows these bubbles 34 to pass through so that they can reach the surface of the reaction liquid 27 unhindered. In addition, a vertical pin 29 is provided for closing and opening the vent hole 28, the vertical pin 29 passing through a corresponding hole in the cover plate 10 so that it can be operated from outside the reaction chamber 9. During the electrophoresis process, the vertical pin 29 closes the vent hole 28 so that no current can flow past the sample 2 through the vent hole 28.
[0063] FIG. 8 shows an electrophoresis device 1 according to a further vertical embodiment, in a vertical cross-section. In contrast to the embodiment shown in FIG. 7, according to the electrophoresis device in FIG. 8 it is provided that the receiving profile 24 receives the sample cassette 19, or the cassette holder 20, at an inclination relative to the horizontal central axis B. The vent hole 28 in this case is realized at the highest point of the cassette holder 20, the highest point being understood to be that region of the cassette holder 20 that is closest to the surface of the reaction liquid 27. Advantageously, the gas bubbles 34 produced when the lower reaction chamber 26 is filled with buffer 27 collect in the vicinity of the vent hole 28, through which they can be discharged toward the surface of the reaction liquid 27.
[0064] FIG. 9 shows a preferred embodiment of the sample cassette 19. The sample cassette 19 comprises a base element 21 and a cover element 22 that can be folded together by means of hinges to form a cassette enclosing the sample 2. According to the preferred embodiment shown, it is provided that the sample cassette 19 is perforated, at least in sections, in particular the base element 21 and the cover element 22 each having a multiplicity of perforations 23 arranged in a grid-like manner. The perforations 23 allow the reaction liquid 27 to reach the tissue sample 2, which in turn ensures that the electric current removes the desired substances from the sample 2. To facilitate the assignment of different samples 2, it is also provided that the sample cassettes 19 have a bar code 35 by which they can be identified. Finally, it may be provided that the sample cassette 19 is produced from a chemically inert and electrically insulating material, in which case polyoxymethylene may preferably be used as the insulating material.
[0065] The invention is not limited to any of the above-described embodiments, but may be varied in a variety of ways.
[0066] All of the features and advantages, including constructional details, spatial arrangements and method steps given by the claims, the description and the drawing, can be essential to the invention both individually and in the widest variety of combinations.
