DE202004009793U1 - Biological reaction chamber for e.g. micro-arrays, microtitration plates and slides, includes permanent magnets in lid and walls - Google Patents

Abstract

A biological reaction chamber comprising permanent magnets in the lid and walls, is new. A biological reaction chamber comprising permanent magnets in the lid and walls, is new. Magnetic subassemblies (11) are fitted in opposite walls of the reaction chamber (1). These align flush with the chamber wall on the cover side, or are blocked off and slightly set back from it. Similarly, in the cover (8), in corresponding positions, second magnetic subassemblies (811) are fitted flush or set back and blocked off, on the side facing the reaction chamber. The first- (11) and second- (811) magnetic assemblies have opposite magnetization. The magnetic subassemblies comprise permanent magnets. There are handling depressions (4) in opposite walls of the reaction chamber, reaching down to its base. The underside of the reaction chamber has projecting magnetic feet (12). External dimensions of the feet correspond with normal microtitration plate formats. Outer walls of the reaction chamber have handle sections (16).

Description

The invention relates to a reaction chamber for Processing, in particular hybridizing, of biological samples, especially those on microarrays, microtiter plates and slides for tent biological Applications.

Microarrays play a key role modern bioscience. Slides are made from microarrays different substrates such as Understanding glass or plastics on which spots of DNA or RNA - oligonucleotides, enzymes, Proteins, etc. can be immobilized. The spots are in one two-dimensional grid arranged on the substrate and in the Usually immobilized. Subsequently the microarray is hybridized, i.e. biological samples who often with Fluorescent markers are marked, applied to the microarray and reaction conditions are created that bind the samples to the Support spots on the array.

Microarrays have been used in research established as a successful tool. For hybridizing the arrays Several devices and procedures are known. A method to perform a hybridization reaction after application place a cover slip on the microarray of the biological sample. The cover glass distributes the sample solution. In addition, the sample solution spreads due to of capillary forces well under the coverslip. Such designs have a disadvantageous effect the strong adhesion of the cover slip to the microarray, the possible inclusion from air bubbles and the low reproducibility.

To avoid this disadvantage, after so-called hybridization chambers used in the prior art, where micro arrays are placed in the chamber, which is covered with a reactant and subsequently be closed with a lid.

The present invention comes closest to in DE 203 01 279 U1 described reaction vessel. The hybridization chamber described there is a reaction container for two or more microarrays. The reaction spaces are sealed to the outside by means of a rubber seal and transparent cover. After this solution, the lid is pressed onto the reaction container using a lid pressing device (screw with pressure cross). A spring mechanism, which has a certain susceptibility to malfunctions and is particularly troublesome during cleaning steps, is used to remove the cover without force.

Similar Hybridization chambers as described above are also used for one or multiple microarrays from companies such as Scienion AG, HTI-Bio-X GmbH offered. There too the chamber is covered by a floor and formed a lid. In the bottom is a holder for the microarray intended. Furthermore, the bottom is covered by a rubber seal, which surrounds the entire array. The lid is placed on the sealing ring and attached to the floor with four screws. This forms there is an airtight seal between the lid, seal and base Chamber in which the microarrays are located.

For optimal execution of a Hybridization reaction become the hybridization chambers described above placed in a tempered immersion bath. The temperature range is depending on the protocol used, between 10 ° C and 100 ° C in the chambers when cooling a negative pressure. This lasts the lid even after loosening all fastenings firmly connected to the ground. To solve the Lid must have adequate pulling force or sideways pull a greater force be applied to lift off the lid. This often leads to Shed the liquid it contains. Furthermore, pulling it sideways can injure the Seal come. The one above force necessary to open the lid is for hinders the user. Another disadvantage is the small number of simultaneously processable microarrays, microtiter plates or slide for tent biological or microarray applications. The type of device design Known and comparable solutions to the present invention further guarantees not the possibility the mixing of the biological sample on the array. This makes fewer signal strength to be expected when evaluating the individual experiments. All the known devices for the special purpose is more for a smaller laboratory scale and manual tests designed.

The object of the invention is a Create reaction chamber for processing biological samples, that are easy and effortless to open can and at the same time a higher sample throughput Use commercially available equipment for processing microtiter plates, e.g. vibrating tables, allows.

This task is done with a reaction chamber solved with the features of claim 1. Advantageous further training are by the subclaims detected.

A reaction chamber according to the present invention has a reaction space in which holders for slides and microarrays or microtiter plates or holders for reaction tubes can be used. In the bottom area of the reaction space, at least one recess for receiving liquids is embedded. Furthermore, the reaction chamber has a lid which can be placed on and connected to the reaction space. A seal is attached between the reaction chamber and the lid. Furthermore, the reaction chamber and the lid are provided with magnetic assemblies which are attached in such a way that their forces act in opposite directions. Reaction chamber and Lids are connected to one another using suitable locking means, for example screws. This creates a closed reaction space that protects the content from external influences.

Repelling has an advantageous effect magnetized components on opening the lid. After hybridization, it can be used despite the prevailing Vacuum is gentle on energy and materials and uncomplicated reaction vessel remove.

Feet are also preferred under the reaction chamber made of magnetic materials. These are attached that the reaction chamber on commercially available devices for systems in microtiter plate format, such as. Microplate shaker or thermal shaker, set up and over the feet mechanically and / or also about the acting magnetic force can be fixed. Has an advantageous effect the shaking of the reaction container on the even distribution the biological sample on the microarray. This leads to a higher number of bonds on the array.

The invention is exemplary Hand the following schematic drawings explained in more detail. Show it:

1 a perspective view of an embodiment of the reaction chamber according to the invention;

2 a plan view of a reaction chamber after 1 ;

3 a perspective view of a lid associated with the reaction chamber and

4 a perspective view of the underside of a reaction chamber according to the invention.

1 shows a perspective view of an embodiment of a reaction chamber according to the invention. A reaction container according to the invention 1 includes a reaction space 2 , The reaction space 2 is dimensioned so that it holds at least one microtiter plate 15 , in the 1 is shown lying on the floor of the reaction space, guaranteed in the format according to ANSI / SBS 01-2004. A plurality of microtiter plates can be used one above the other, which are spaced apart from one another by a peripheral frame (not shown). It is within the scope of the invention, mounts 3 to provide the in 1 are indicated in a partial section, which enable the side-by-side recording and simultaneous spacing of several sample holders in microtiter plate format. To simplify loading and unloading, there are at least two opposite walls of the reaction space 2 intervention troughs 4 intended. According to the invention are at least in two opposite wall areas of the reaction container 1 first magnetic assemblies 11 introduced, which are flush with the chamber wall or slightly behind. These magnetic assemblies are advantageously formed by cylindrical permanent magnets, without the invention being limited to such an embodiment. In the example, the magnets are pressed in and if necessary cast flush.

In the top edge of the reaction container 1 a circumferential seal is advantageous 5 used. Through the seal 5 is used in conjunction with a lid 8th (see. 3 ) the reaction space 2 sealed when closing means, preferably in the form of through the lid 8th through holes 9 in threaded holes 10 screws engaging the housing wall 14 be introduced. The reaction space 2 (see. 2 ) is in the ground 6 with at least one depression 7 provided for the absorption of liquids. Depending on the application, several recesses are also possible.

In the example are on the two transverse sides of the lid 8th two magnetized elements each 811 , for example permanent magnets, introduced as in 3 indicated. These magnets 811 correspond in their position with those in the walls of the reaction container 1 inserted magnets 11 , but have opposite polarization. For the magnets in the lid 8th have nickel-plated neodymium-iron-boron magnets with a diameter of 6 mm and a height of 4 mm and a magnetization of 1300 mT [Millitesla] and for the magnets 11 in the chamber walls, those with a diameter of 4 mm and a height of 12.5 mm and such magnetization prove to be advantageous. Such a design guarantees problem-free opening of the reaction chamber under reduced pressure after the experiment has been carried out with a chamber size of the order of 100-150 mm by pushing off the lid. Other chamber sizes require more or fewer magnets or different geometries of the magnets or, if necessary, the provision of yokes.

The arrangement of feet is also within the scope of the invention 12 on the underside of the floor 13 of the reaction container 1 (see. 4 ) intended. As feet 12 For example, suitable permanent magnets can be used. These are attached in such a way that their external dimensions correspond to the standard microtiter plate formats. Due to these dimensions, the reaction chamber can be used on almost all commercially available devices for microtiter plates. In contrast to known chambers, this ensures, for example, active mixing by means of a shaker and thus a more uniform binding of the samples on the microarray.

In the context of the invention, it is also advantageous to use the opposite longitudinal sides of the outer walls of the reaction container 1 with grips 16 , especially in the form of recessed grips.

1

reaction vessel

2

reaction chamber

3

bracket

4

engagement troughs

5

poetry

6

ground

7

deepening

8th

cover

9

drilling

10

threaded holes

11

first permanent magnets

811

second permanent magnets

12

feet

13

Bottom underside

14

closure means (Screws)

15

microtiter plate

16

Handle (recess)

Claims (9)

Reaction chamber for processing biological samples, especially those on microarrays, microtiter plates and slides, consisting of a reaction container ( 1 ) with at least one reaction space ( 2 ) for holding at least one sample holder, an attachable lid ( 8th ), a seal ( 5 ) between reaction vessel ( 1 ) and lid ( 8th ) and closure means ( 14 ), characterized in that at least in two opposite wall areas of the reaction container ( 1 ) first magnetic assemblies ( 11 ) are inserted, which are flush with the chamber wall on the cover side or slightly behind and in the cover ( 8th ) with the first magnetic assemblies ( 11 ) corresponding second magnetic assemblies ( 811 ), which is also flush or slightly behind the reaction vessel ( 1 ) facing the cover side are introduced in such a way that the first ( 11 ) and second magnetic assemblies ( 811 ) have an opposite magnetization. Reaction chamber according to claim 1, characterized in that the magnetic assemblies ( 11 ; 811 ) are formed by permanent magnets. Reaction chamber according to claim 1, characterized in that the reaction space is dimensioned so that it at least is able to hold a microtiter plate in ANSI / SBS 01-2004 format. Reaction chamber according to claim 1 or 3, characterized in that in at least two opposite walls of the reaction container ( 1 ) Intervention troughs ( 4 ) are introduced. Reaction chamber according to claim 4, characterized in that the engagement troughs ( 4 ) to the bottom of the reaction container ( 1 ) extend. Reaction chamber according to claim 1, characterized in that the bottom underside ( 13 ) of the reaction container ( 1 ) with raised feet ( 12 ) is provided. Reaction chamber according to claim 6, characterized in that the feet ( 12 ) are magnetizable or formed by magnets. Reaction chamber according to claim 6 or 7, characterized in that the feet ( 12 ) are arranged in an external dimension that corresponds to the footprint of conventional microtiter plate formats. Reaction chamber according to claim 1, characterized in that preferably the opposite longitudinal sides of the outer walls of the reaction container ( 1 ) with grips ( 16 ) are provided.