• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Protective element of aqueous samples in thermocyclers, designed to interpose between dropchips (1) containers of aqueous samples arranged on the surface of a tray (2) located inside a PCR thermocycler (3) and a cover (4) of said thermocycler (3). The protective element comprises a lower sheet (5) of material with thermal insulation capacity, designed to face the dropchips (1), with a first central window (7), in a central sector and first perimeter windows (8), and an upper sheet (6) of material with resistance to deformation, intended to be interposed between the lower sheet (5) and the cover (4), upper sheet (6) which in turn has a second central window (9) intended to face the first central window (7), and second perimeter windows (10), intended to face the first perimeter windows (8). (Machine-translation by Google Translate, not legally binding)
    公开号:ES2703981A1
    申请号:ES201731112
    申请日:2017-09-13
    公开日:2019-03-13
    发明作者:Martinez Ana Belen Rodriguez；Barcina Maria Garcia；Diez Esther Sarasola
    申请人:Administracion General de la Comunidad Autonoma de Euskadi；
    IPC主号:
    专利说明:

    [0001] PROTECTIVE ELEMENT OF AQUEOUS SAMPLES IN THERMOSCYCLES
    [0002]
    [0003]
    [0004]
    [0005] OBJECT OF THE INVENTION
    [0006]
    [0007] The present invention falls within the technical field of laboratory apparatus for heating or cooling, more specifically in those devices specifically adapted for the treatment of nucleic acid samples, as well as in the additional construction details of said apparatuses, and refers in particular to a sheet-like protective element for coating and protecting container chips from aqueous nucleic acid samples inside thermal cyclers.
    [0008]
    [0009] BACKGROUND OF THE INVENTION
    [0010]
    [0011] A thermal cycler, also known as a PCR machine, is a device used in molecular biology that allows to perform the temperature cycles necessary for a chain reaction of the DNA amplification polymerase. The most common thermocycler model consists of an electrical resistance block that distributes a homogeneous temperature through a plate during pre-established times, normally with approximate temperature ranges from 4 ° C to 96 ° C.
    [0012]
    [0013] Since the reactions incubated in the apparatus are on aqueous solutions, the thermal cyclers usually include a cover plate constantly heated to approximately 103 ° C, to avoid condensation of the water in the caps of the tubes or sample containers where the reaction occurs, and thus prevent the solutes from concentrating, which would modify the optimal conditions for the polymerizing enzyme and the thermodynamics of the primer pairing.
    [0014]
    [0015] Prior to its introduction inside the thermal cycler, liquid samples with DNA, conveniently mixed with a solute, are introduced into the sample container to be centrifuged. On the other hand, after carrying out the amplification of DNA inside the thermal cycler, we proceed to the scanning and subsequent reading of the image obtained to obtain and interpret the results.
    [0016]
    [0017] In the current state of the art, support plates containing samples, also known as dropchips (1), are intended to house at least two samples of genetic material in liquid state. As seen in Figure 1, each support plate comprises a laminar body (101) of essentially rectangular geometry and made of plastic materials, which has an upper face (102) in which two modules (103) are located for housing of samples.
    [0018]
    [0019] Each of said modules (103) in turn comprises an entrance chamber (104) provided with a hole through which the diluted aqueous sample is introduced, wherein said orifice is covered by an aeration membrane (105). From said inlet chamber divides a duct (106) in the form of a circuit, inside which the samples are divided into a plurality of drops during the emulsion centrifugation process.
    [0020]
    [0021] The duct (106) ends in a reading chamber (107) inside which the droplets formed after the centrifugation are deposited to carry out the PCR reaction and, subsequently, to read the results by scanning with a microarray scanner. Said reading chamber (107) must therefore be left free to perform the scanning.
    [0022]
    [0023] This type of dropchips (1) are usually placed on the surface of a tray located inside the thermal cycler, so that up to four dropchips can be placed on a single tray. Said tray keeps them in a suitable position so that the heat transfer cap proper of the thermal cycler comes into contact with the dropchips (1), so that inside the reading chamber (107) the necessary temperature is reached so that produce the chain reaction of the polymerase that amplifies the DNA.
    [0024]
    [0025] As indicated above, the thermocyclers available in the market have a flat lid that, when closed, is in direct contact with the sample tubes. In the case of the dropchips (1), the contact is practically complete with the entire surface of its upper face (102), which is not beneficial for the performance of the technique, since it physically affects its structural integrity and may be deformed and the sample itself, which may even cause the evaporation of it. The curvature that causes the loss of structural integrity of the dropchip (1) prevents establishing a uniform focal plane of the entire surface to analyze the results by means of an optical reader.
    [0026]
    [0027] Experiments carried out have shown that the most optimal way to maintain the dropchip (1) without curvature is to have a protective element that leaves free the zones of the dropchip (1) corresponding to the reading chamber (107) and the membrane (105). ) and that is at the same time capable of exerting a pressure on the perimeter edges of the dropchip (1) to prevent it from curling after the thermocycling.
    [0028]
    [0029] DESCRIPTION OF THE INVENTION
    [0030]
    [0031] The object of the invention consists of a protective element designed to interpose between some dropchips placed on the surface of a tray located inside a PCR thermocycler of the type currently existing and the cover of said thermal cycler, to prevent the dropchips they deform and curl due to the heat transmitted directly by the lid, thus facilitating the reading of results by scanning.
    [0032]
    [0033] For this, the element comprises at least two laminar bodies of similar geometry and dimensions superimposed on each other. The first laminar body constitutes a lower layer of the protective element, and is intended to be arranged in direct contact with the dropchips, for which reason it is made of a material with a capacity for damping the heat transfer, preferably silicone.
    [0034]
    [0035] The second laminar body constitutes an upper layer of the element, and is intended to be arranged in direct contact with the cover of the thermal cycler, so it is made of a material with good capacity for structural resistance to deformations produced by heat, preferably brass.
    [0036]
    [0037] The main advantages of the protective element lie in the fact that it allows the execution of the technique in any thermal cycler, since it avoids deformation of the dropchip, allowing its reading by scanning, as well as avoiding the evaporation of the samples housed inside said dropchip. Likewise, the protective element thus described allows simultaneous thermocycling of up to four dropchips arranged on the thermocycler tray.
    [0038]
    [0039] DESCRIPTION OF THE DRAWINGS
    [0040]
    [0041] To complement the description that is being made and in order to help a better understanding of the characteristics of the invention, according to a preferred example of practical realization thereof, a set of drawings is attached as an integral part of said description. where, with illustrative and non-limiting character, the following has been represented:
    [0042]
    [0043] Figure 1 shows a plan view of a dropchip according to the current state of the art.
    [0044]
    [0045] Figure 2.- Shows a plan view of an exploded view of the protective element according to a first preferred embodiment, in which its main constituent elements are appreciated.
    [0046]
    [0047] Figure 3.- Shows a plan view of an exploded view of the protective element according to a second preferred embodiment.
    [0048]
    [0049] Figure 4 shows a perspective view of the interior of a PCR thermal cycler.
    [0050]
    [0051] Figure 5 shows a perspective view of the interior of the thermal cycler with a plurality of dropchips disposed on the tray.
    [0052]
    [0053] Figure 6 shows the view of figure 5 with the lower layer of the protective element arranged on the dropchips.
    [0054]
    [0055] Figure 7 shows the view of figure 6 with the upper layer of the protective element of the first preferred embodiment arranged overlying the lower layer.
    [0056] PREFERRED EMBODIMENT OF THE INVENTION
    [0057]
    [0058] A detailed explanation of a preferred embodiment of the object of the present invention is given below, with the aid of the aforementioned figures 2 to 7.
    [0059]
    [0060] The protective element of aqueous samples in thermocyclers described is intended to be interposed between an upper face (102) of the laminar body (101) of dropchips (1) containers of aqueous samples placed on the surface of a tray (2) located in the inside of a PCR thermocycler (3) and a cover (4) of said thermocycler (3), to prevent the laminar bodies (101) from deforming and curling due to the heat transmitted by the cover (4), thus facilitating the reading results by scanning.
    [0061]
    [0062] To do this, the protective element is formed by a lower sheet (5), intended to be faced and in direct contact with the dropchips (1) arranged on the tray (2), and an upper sheet (6), of geometry and dimensions perimetral essentially similar to those of the lower sheet (5), to which it overlaps, said upper sheet (6) being intended to face and in direct contact with the cover (4) of the thermal cycler (3). In an alternative embodiment, the lower (5) and upper (6) sheets are fixed to each other.
    [0063]
    [0064] As seen in figures 2 and 3, the lower sheet (5) has an essentially polygonal geometry, more specifically rectangular, and dimensions essentially coinciding with those of the tray (2). Said lower sheet (5) is made of materials with thermal insulation capacity, to avoid an excess of direct transfer of heat from the cover (4) to the dropchips (1), which could produce deformations in the dropchip itself (1). ), as well as getting to evaporate the samples in liquid state housed inside. In the preferred embodiment described herein, the lower sheet (5) is made of silicone.
    [0065]
    [0066] The lower sheet (5) also has a plurality of through openings in the shape of a window. A first central window (7), located in the central area of said lower sheet (5) and of essentially rectangular geometry, avoids coating of the zone of the dropchip (1) in which the membranes (105) are located, which allows the aeration of the sample avoiding in this way that they lose their integrity and even evaporate. First perimeter windows (8), located along two opposite perimeter edges of the lower sheet (5), are intended to avoid coating the reading chamber (107) of the dropchip (1), to allow direct reading of a sample housed in said reading chamber (107) by a scanner.
    [0067]
    [0068] In the preferred embodiment described herein, the first perimeter windows (8) have an essentially square geometry and dimensions essentially similar to those of the reading chamber (107) of the dropchip (1), and the lower sheet has a total of eight first Perimeter windows (8), which allows the coating of up to four dropchips (1). These first eight perimeter windows (8) are grouped in an upper row (801) and in a lower row (802) with respect to the first central window (7).
    [0069]
    [0070] Figures 2 and 3 illustrate two respective embodiments of the upper sheet (6), which has a geometry and dimensions essentially similar to those of the lower sheet (5), on which it is intended to overlap. The upper sheet (6) is made of materials with a capacity of resistance to deformation by temperature, to avoid deformations due to the heat transmitted by the cover (4). In this preferred embodiment, the upper sheet (6) is materialized in brass, although the option of making it in various metallic materials or its alloys is contemplated.
    [0071]
    [0072] The upper sheet (6) also has a second central window (9), intended to face the first central window (7) of the lower sheet (5) to allow air to enter through the membrane (105), as well as it presents second perimeter windows (10), intended to face the first perimeter windows (8) of the lower sheet (5) to allow a space between the cover (4) and the reading chamber (107), thus avoiding an excess in the application of pressure on it and consequently, preventing the evaporation of the aqueous sample.
    [0073] The second perimeter windows (10) are grouped in a first column (1001) of parallel windows superior to the second central window (9) and in a second column (1002) of parallel windows inferior to the second central window (9).
    [0074]
    [0075] Figure 2 illustrates a first preferred embodiment of the protective element, in which the second perimeter windows (10) have an essentially square geometry and dimensions, similar to those of the first perimeter windows (8) of the lower sheet (5), which are destined to face.
    [0076]
    [0077] Figure 3 shows a second preferred embodiment of the protective element, in which the second perimetric windows (10) are rectangular, so that when superimposed on the first perimeter windows (8) they create a kind of grid.
    [0078]
    [0079] Figure 4 shows how a tray (2) is coupled to the interior of a thermocycler (3), which will then be covered by the cover (4) that will transmit heat. The tray (2) is designed to house up to four dropchips (1), arranged as shown in figure 5, with their respective input chambers (104) located in the central area of the tray (2), and reading chambers (107) arranged in the vicinity of two opposite perimeter edges of said tray (2).
    [0080]
    [0081] In figure 6 it can be seen how the lower sheet (5) of the protective element covers the dropchips (1) arranged on the tray (2), leaving the respective reading cameras (107) and membranes (105) of their cameras free. of entry (104). Finally, Figure 7 shows how the upper sheet (6) overlaps the lower sheet (5) to be in direct contact with the cover (4) of the thermal cycler (3) when it closes on the tray (2) and the dropchips (1).
    [0082]
    [0083] The protective element of aqueous samples in thermocyclers exerts a uniform and balanced pressure on the dropchips (1) as well as avoids the direct transfer of heat from the cover (4) of the thermocycler (3) during the thermocycling procedure of the samples housed in the modules (103) of the dropchips (1), which avoid alterations in the sample and in the integrity of the dropchip itself (1), which allows a correct reading of the results obtained.
    权利要求:
    Claims (10)
    [1]
    1. Protective element of aqueous samples in thermal cyclers to interpose between dropchips (1) containers of aqueous samples arranged on the surface of a polygonal tray (2) located inside a PCR thermocycler (3) and a lid (4) ) of said thermocycler (3) heat transmitter, to prevent direct heat transfer of the cover (4), in which each of said dropchips (1) has a laminar body (101) of plastic materials, with one face upper (102) in which two modules (103) for locating samples are located, the protective element being characterized in that it comprises:
    - a lower sheet (5) of material with thermal insulation capacity and geometry and dimensions essentially similar to those of the tray (2), intended to face and in direct contact with the dropchips (1) arranged on said tray (2) , lower sheet (5) that presents in turn:
    - a first central window (7), located in a central sector of the lower sheet (5), and
    - first perimeter windows (8), and
    - an upper sheet (6) of material with a capacity of resistance to deformation by temperature and of geometry and dimensions essentially similar to those of the lower sheet (5), intended to be interposed between the lower sheet (5) and the cover (4) ), upper sheet (6) which in turn presents:
    - a second central window (9) intended to face the first central window (7), and
    - second perimeter windows (10), designed to face the first perimeter windows (8).
    [2]
    2. A protective element for aqueous samples according to claim 1, characterized in that the first central window (7) and the second central window (9) have an essentially similar geometry and dimensions.
    [3]
    3. Protective element for aqueous samples according to claim 1 or 2, characterized in that the first perimeter windows (8) and the second perimeter windows (10) have an essentially similar geometry, dimensions and distribution.
    [4]
    4. Protective element of aqueous samples according to claims 1 or 2 characterized in that the first perimeter windows (8) are grouped in an upper row (801) and in a lower row (802) with respect to the first central window (7) .
    [5]
    5. A protective element for aqueous samples according to claim 1 or 2, characterized in that the second perimeter windows (10) are grouped in a first column (1001) of parallel windows, higher than the second central window (9), and in one second column (1002) of parallel windows, lower than the second central window (9).
    [6]
    6. Protective element of aqueous samples according to claim 5 characterized in that the second perimeter windows (10) are rectangular.
    [7]
    7. Protective element of aqueous samples according to claim 4, characterized in that the first perimeter windows (8) are square.
    [8]
    8. Protective element of aqueous samples according to any of the preceding claims, characterized in that the lower sheet (5) is made of silicone.
    [9]
    9. Protective element of aqueous samples according to any of the preceding claims, characterized in that the upper sheet (6) is made of brass.
    [10]
    10. A protective element for aqueous samples according to any of the preceding claims, characterized in that the lower sheet (5) and the upper sheet (6) are fixedly linked to each other.
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    引用文献:
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    US20060246580A1|2005-05-02|2006-11-02|Jin-Tae Kim|Polymerase chain reaction module, and multiple polymerase chain reaction system including the module|
    EP1980321A2|2007-04-09|2008-10-15|Hitachi Software Engineering Co., Ltd.|Reaction apparatus and reaction chip|
    US20090148912A1|2007-12-06|2009-06-11|Seiko Epson Corporation|Biological sample reaction chip, biological sample reaction apparatus, and biological sample reaction method|
    US20090186404A1|2008-01-22|2009-07-23|Samsung Electronics Co., Ltd.|Apparatus for amplifying nucleic acids|
    US20150190811A1|2012-06-22|2015-07-09|Panasonic Intellectual Property Management Co., Ltd.|Microfluidic device|
    法律状态:
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    优先权:
    申请号 | 申请日 | 专利标题
    ES201731112A|ES2703981B2|2017-09-13|2017-09-13|PROTECTIVE ELEMENT OF AQUEOUS SAMPLES IN THERMOCYCLERS|ES201731112A| ES2703981B2|2017-09-13|2017-09-13|PROTECTIVE ELEMENT OF AQUEOUS SAMPLES IN THERMOCYCLERS|
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