Patent Application
20080032034
The present invention is explained in greater detail below with reference to the attached drawings and to preferred exemplary embodiments. In the drawings:
On a lower side of the supporting element 3 there is a receiving surface for in particular biological specimens, which is covered with an agarose layer 4 preferably consisting of high-quality, i.e. high purity LE agarose. The agarose layer serves as an adhesive agent for securing the specimens to the receiving surface. In principle, another suitable hydrogel or another adhesive agent with the desired properties may also be used instead of the agarose layer 4.
For the catapulting process described in detail with reference to
For further processing, the receiving element 1 is then placed for example on a cell culture vessel or indeed on a further Petri dish 5 with membrane 5a. The cells catapulted onto the hydrogel 4 may be detached by gentle motion or by heating the agarose layer 4. The further Petri dish 5 may in particular be filled with a cell culture liquid, into which the hydrogel 4 dips. Alternatively, the hydrogel layer may also be completely dissolved by the addition of agarase, an enzyme which dissolves agarose. Then the receiving element 1 may be replaced with a conventional lid and reused, optionally after sterilisation. If the catapulted cells are transferred in this way into a further Petri dish 5 with membrane 5a, the process may be repeated after culturing in this Petri dish with the cell cultures which arise therein.
Use of the hydrogel, in this case the agarose layer 4, has the advantage that electrostatic forces produced for example by the laser irradiation or by contact do not result in the cells which have been catapulted out remaining attached to the lid 2 instead of on the hydrogel layer 4.
Another advantage of using such a receiving means is that the agarose layer 4 allows better visualisation of the cell culture 7 and the catapulted-out cells with a microscope, since contrast is improved.
After catapulting, the agarose 4 may again be liquefied by the addition of agarase, such that immediate re-use is possible. Moreover, different additives such as for example cell culture media or buffer solutions may be introduced in the agarose layers 4. For subsequent processing or analysis of the catapulted cells, additional introduction of these agents is therefore no longer necessary, since they are liberated once the agarose has been dissolved or liquefied. Enzymes or enzyme prebatches may in this case also serve as additives. One operation is therefore dispensed with, so avoiding the associated risk of contamination, since these additives do not have to be added for example by pipetting, and the desired processing and/or analysis may be initiated immediately. One preferred field of application is in this case the polymerase chain reaction (PCR), wherein the additives may preferably be so selected that the reaction is initiated at the same time as dissolution of the hydrogel by heating. Another field of application is culturing of the catapulted cells.
In such applications, care must be taken to ensure that the dissolved hydrogel does not interfere with or contaminate the intended processing or analysis. High purity, preferably sterilised agarose is here too an example of a suitable hydrogel.
Instead of multiple culture dishes, so-called microtitre plates or 96-well microtitre plates may also be filled, as in
It goes without saying that the exemplary embodiments described here are not limited to agarose as hydrogel, it also being possible to use other hydrogels with the desired properties. For example, a hydrogel based on collagen, polyacrylamide or similar substances would be possible here. The receiving units may also assume different forms, depending on the desired application.
| Number | Date | Country | Kind |
|---|---|---|---|
| 103 58 565.6 | Dec 2003 | DE | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP04/14311 | 12/15/2004 | WO | 00 | 3/26/2007 |
