DE102009048039A1 - Sample e.g. biological sample, treatment device for use in e.g. laboratory to remove undesirable constituent, has sample units connected in row, where sample volumes are connected in locking bar area so that gap is developed between volumes - Google Patents

Abstract

The device has a locking bar (1) in which eight sample units (8) are connected in a row by a bar (4). Each sample unit comprises membranes (5) and an opening (6) for extraction and addition of samples. Sample volumes (2) in a capillary form are connected only in an upper locking bar area by the bar, so that a gap (3) is developed between the sample volumes. The locking bar comprises two projections (7) on front and rear sides, which the projections are perpendicular to a locking bar plane. Each projection comprises a rounded-off or angled lower corner.

Description

The invention describes a device compatible with the microtiter plate format and the frequently used microcentrifuge tubes. When working in the medical, molecular biological, biotechnological, pharmaceutical, chemical or environmental analytical laboratory, frequently fluctuating sample numbers occur. The device is used for more effective and easier processing of very different numbers of samples of liquid samples, especially in aqueous solutions. These are z. For example biological or medical samples in the sample preparation or treatment in the volume of 2 to 150 μl. The simple production can be a one-time use as a disposable product, which saves cleaning and reduces the risk of contamination. Biological but also other samples often contain accompanying components that are to be selectively removed. For this purpose, the different properties of the accompanying components such. Eg size, charge or hydrophobicity used.

An example of sample processing is the denaturation of a protein in a denaturing agent, such as urea. The denaturing agent must be removed for subsequent analysis steps. For this purpose, a separation of the low molecular weight urea from the much higher molecular weight protein by diffusion through a membrane with pores of a size that allows the urea to pass and retain the protein can be used. For this purpose, the sample is placed in a sample unit ( 8th ), with the semipermeable membrane ( 5 ) as part of at least one boundary of the sample unit ( 8th ). The sample unit ( 8th ) is placed with the sample in an outer container of a solution z. Eg contains a buffer and is referred to below as a buffer vessel. The membrane in this case represents a selective contact between the sample and the solution or the buffer. This principle is present in the device described and can of course. also be used for the separation of other high molecular weight biopolymers such as sugar polymers, DNA or RNA of low molecular weight components. Alternative methods of separating the exemplified substances would be ultrafiltration methods, chromatography methods such as gel chromatography and precipitation methods. Further possibilities of use of the device described here are the concentration and rebuffering of samples as well as the carrying out of sequential enzymatic reactions in a sample vessel or the selective supply or removal of substances to cell cultures in a sample vessel.

State of the art:

For the sample treatment according to the principle of diffusion through selective membranes, there are different solutions (eg. DE 101 60 975 A US 2004195163 . US 2005019774 . US 2005148066 . US5503741 , SpectrumLabs: Spectra / PorMicrodialyzer).

Also on the market there are various offered solutions for disposable products, which can be distinguished among other things according to the number of samples to be processed in parallel. Primarily for single samples, products such as the Float-A-Lyzer, Slide-A.Lyzer (Pierce), Tube-O-Lyzer (Geno Technology), BioDispoDialyser (Spectrum) are suitable for different volumes. For parallelized processing of samples there are 96-well dialysers (Harvard Bioscience) and D-Tube96 ^™ Dialyzer (EMD Bioscience) with 96 sample places.

An intermediate solution in terms of the number of samples offers DE 10 2007 011 866 A1 with a cartridge with a bar-shaped structure, hereinafter referred to as dialysis lock, with preferably 8 sample sites with sample volumes of 1-500 .mu.l and can be built with in the grid of the microtiter plates. This allows samples within the microtiter plate grid z. Eg be transferred with 8-fold pipettes. The solution also has a particularly high dialysis rate through the capillary geometry of the sample vessels and membrane. However, it can not be used in such a way that the bars can be placed individually with each sample space in an outer buffer vessel. Each bar comes in a common buffer vessel. Thus, an exchange of substances across the membrane, the solvent in the buffer from a sample vessel reach into adjacent sample vessels. All sample containers, with or without sample, are placed in an external buffer.

The invention is based on the object to provide a device which is based on a known latch-shaped device for sample processing ( DE 10 2007 011 866 A1 ). The user can flexibly process very different sample numbers with the proposed solutions, without any additional expenditure on equipment and work, and use the standard equipment in the laboratory and standard consumables.

In the following, a particularly advantageous embodiment is illustrated by means of an embodiment.

The device is in 1 represented and consists of a bolt ( 1 ) in the 8 sample units ( 8th ) in a row with each other by a bridge ( 4 ) are connected. Each sample unit exists from the membranes ( 5 ), the sample volume in capillary form ( 2 ), Opening for sampling and addition ( 6 ), Pressure compensation and overflow ( 10 ), Head Start ( 7 ), Plate ( 11 ) and surfaces ( 12 ). The opening for sampling and addition ( 6 ) are conical inside and in their dimensions the reversible densities with conventional pipette tips z. For example, the volumes 2-200 ul and 100-1000 ul adjusted. This allows the sample in the volume of 5-110 ul by pressing the pipette with slight overpressure from the pipette tip into the U-shaped sample volume in capillary form ( 2 ) to be brought. It is useful, with the excess volume of the pipette by repressing air, the position of the liquid column within the sample volume in the capillary ( 2 ) to achieve optimal results. Within a bar, up to 8 samples can be accommodated in parallel. For evaporation and contamination protection, the opening for sampling and addition ( 6 ) and pressure compensation and overflow ( 10 ) with a strip of adhesive tape. The bars will be like in 2 shown in a standard deep well microtiter plate ( 13 ). The individual volumes of the microtiter plate in this case contain up to 2000 ul of a solution z. Eg a dialysis buffer. Typically, z. For example, used to desalt or rebuffer 1800 ul. This ensures a ratio of the volume of dialysis buffer to sample of 1: 16.4 at maximum sample volumes (110 μl) and 1: 360 at minimum sample volumes (5 μl). Within a deep well microtiter plate ( 13 ) can be 1 to 12 bars ( 1 ) are used simultaneously. Will less bars ( 1 ) in a deep well microtiter plate ( 13 ), equal or different gradations of the concentration of a solution but also different solutions in the volumes of a deep well microtiter plate ( 13 ) submit. In this the bar ( 1 ) are easily transferred with the samples after the first part of the sample processing. As a result, sequential processes such as those frequently used in multistep processes or in enzymatically catalyzed reactions, for example, can be used. For example, occur the tryptic protein digestion, perform in the same sample vessel. The composition of the samples in the sample volume in capillary form ( 2 ) is achieved simply by moving the bar within a deep well microtiter plate ( 13 ) or between several Deep Well microtiter plates ( 13 ) through the selective membrane. Concentrating the sample is possible by the dialysis buffer in the deep well microtiter plate ( 13 ) having a higher osmolarity than the sample, wherein the substance such. For example, higher molecular weight dextran increase the osmolarity in the dialysis buffer due to the high molecular weight of the membrane ( 5 ) can not happen. Will the bar ( 1 ) brought into a gas stream with the samples, the solvent evaporates z. For example, water across the membrane quickly and concentrate the sample. For the use of individual samples, the bridge ( 4 ) provided as a predetermined breaking point or interface. Here, one or more sample units ( 8th ) split off. Individual sample units ( 8th ) are the dimensions of a microcentrifuge tube ( 14 ) with 2 ml sample volume. The single sample unit can thus be transferred without further processing into a 2 ml microcentrifuge tube ( 14 ), which serves as a dialysis buffer vessel, are used and the lid closed. After the end of dialysis or sample treatment, the sample unit ( 8th ) and the sample is taken and subjected to analysis or sequentially into another microcentrifuge tube ( 14 ) are treated with the same or different solutions. If the process for sample processing is to be accelerated, both microcentrifuge tubes ( 14 ) as well as deep well microtiter plate ( 13 ) are moved with a conventional shaker with shaking speeds of 100 to 3500 revolutions per minute. The marking of the bolt ( 1 ), the user can by a label with a smudge-proof pen or by sticking a labeled label on the plate ( 11 ) at the end of the bolt ( 1 ) carry out. The plate ( 11 ) also has a predetermined breaking point, which in the same way as for the separation of individual sample units ( 8th ) is used to separate the plate ( 11 ) is being used. Thus, the associated sample unit ( 8th ) into a microcentrifuge tube ( 14 ) can be used. The name of the individual sample units ( 8th ) can as in on the surface ( 15 ) is exemplified by letters, analogous to the standard designation of the microtiter plates with the letters A-H.

pictures

1 Bar and sample unit for sample processing

2 , Arrangement of the bars in a microtiter plate in deep well format

3 , Sample unit in the microcentrifuge tube

LIST OF REFERENCE NUMBERS

1

bars

2

Sample volume in capillary form

3

gap

4

web

5

membrane

6

Opening for sampling and addition

7

head Start

8th

sample unit

9

imaginary circle

10

Pressure equalization and overflow hole

11

plate

12

area

13

Deep well microtiter plate

14

Micro centrifuge tubes

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• DE 10160975 A [0003]
• US 2004195163 [0003]
• US 2005019774 [0003]
• US 2005148066 [0003]
• US 5503741 [0003]
• DE 102007011866 A1 [0005, 0006]

Claims (11)

Device for sample treatment in small volumes, consisting of a known bar form ( 1 ) with a plurality of vertically arranged sample volumes in capillary form ( 2 ) the membranes which are selective on both longitudinal sides ( 5 ) are arranged at least once curved through 360 ° in a plane in which at least the openings for sampling and addition ( 6 ) above, characterized in that the sample volumes in capillary form ( 2 ) only in the upper bar area by at least one bridge ( 4 ) are connected so that between the sample volumes in capillary form ( 2 ) A gap ( 3 ) arises. Device according to claim 1, characterized in that the web ( 4 ) below the upper edge of the bolt ( 1 ) where the vertical thickness is less than 4 mm, the bridge ( 4 ) narrower than the width of the bolt and the width is greater than 0.5 mm and the web ( 4 ) by a vertical and or horizontal wedge-shaped constriction is a predetermined breaking and / or interface and thus single and / or multiple sample units ( 8th ) are separated at the predetermined breaking / interfaces around the respective desired number of sample units ( 8th ) to obtain. Device according to claim 1, characterized in that the sample volumes are in capillary form ( 2 ) a maximum length of 46 mm, preferably less than 35 mm and a minimum length of 4 mm from the lower edge of the web ( 4 ) to its lower edge and a maximum width of 9 mm and a minimum width of 3 mm and the center ( 10 ) the sample volumes in capillary form ( 2 ) are each 9 mm apart, the gap ( 4 ) between the sample volumes in capillary form ( 2 ) is at least 0.8 mm and at most 5 mm, and thus are suitable for conventional grid dimensions of microtiter plates. Device according to claim 1, characterized in that the latches have at least 2 projections ( 7 ) on the front and rear side, which are perpendicular to the bar plane and allow a centering of the bar in the upper region of the individual sample volumes of sample containers in the microtiter plate format ( 2 ). Device according to claims 1 and 9, characterized in that the projections ( 7 ) have a rounded or sloping bottom corner and allow for easy insertion of the bar at the top of each sample volume of microtiter plate size sample containers ( 2 ). Device according to claim 1, characterized in that the projections ( 7 ) in the middle of the sample volumes in capillary form and the upper part of the horizontal surface ( 12 ) a sample unit ( 8th ), bounded by the bridge ( 4 ) and / or the end of the bolt ( 1 ) in the edge regions, by the chamfered and / or rounded corners including the projection (s) ( 7 ) within an imaginary circle ( 9 ) of maximum 9 mm diameter around the center bar ( 10 ) of a sample volume in capillary form ( 2 ) as the center of the circle, and thus centered in a conventional microcentrifuge tube 3 ) can be used. Device according to claim 1, characterized in that the opening for sampling and addition ( 6 ) in funnel shape with a maximum height of 10 and a minimum height of 1 mm, the lower diameter being between 0.1 and 2 mm preferably between 0.5 and 1 mm and the upper diameter between 1 and 9 mm preferably between 2 and 5 mm, thereby enabling reversible sealing by light pressure with a standard pipette tip for sample addition and removal and / or flushing of sample volumes in capillary form. Device according to claim 1, characterized in that the pressure compensation and overflow holes are made in a funnel shape with a maximum height of 10 mm and a minimum height of 1 mm, the lower diameter being between 0.1 and 2 mm and the upper diameter between 1 and 9 mm and thus form an overflow space for excess sample. Device according to claim 1, characterized in that at one or both ends of the device via a web ( 4 ) as breaking point / interface a small plate ( 11 ) is applied to markings / labels whose width is not greater than 9 mm and whose length is not greater than 12 mm and that at least on one side of the upper side of the bolt ( 1 ) Areas ( 12 ) are provided with which the individual sample units can be marked or labeled. Use of the device according to claims 1-16, characterized in that the device with at least one sample unit successively in different columns or rows of one or more microtiter plates, each with the same or various solutions for removing undesired constituents, supplying desired constituents and or rebuffering the sample according to the selectivity of the membrane ( 5 ) is used. Use of the device according to claims 1-16 and 17, characterized in that a sample unit ( 8th ) successively in different centrifuge tubes, each with the same or different solutions for removing unwanted components, supplying desired components and or rebuffering the sample according to the selectivity of the membrane ( 5 ) is used.

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