Patent Application
20200298241
This U.S. patent application claims priority to European patent application serial no. EP 19163639, filed Mar. 19, 2019 This prior application is incorporated by reference in its entirety.
Not applicable.
Not applicable.
This invention relates to a slide chamber i.e. a device providing wells attached to a microscope inspection slide for analyzing biological specimens.
In immunofluorescence analysis, fluorescent dyes conjugated to one or more antigen recognizing moieties like antibodies are used for specific detection and isolation of target cells. To this end, a vast number of antibodies, fluorescent dyes, flow cytometers, flow sorters and fluorescence microscopes has been developed.
One technique of immunofluorescence analysis makes use of sequential elimination of the fluorescence signal and re-staining of the same biological specimen, thus allowing detection of a plurality of targets (antigens) on the biological specimen resulting in much higher multiplexing potential compared to standard procedures using simultaneous labeling and detection. For example, U.S. Pat. No. 7,741,045 B2, EP 0810 428 B1 or DE10143757 disclose elimination of the fluorescence signal by photo- or chemical destruction of the conjugated fluorescent moieties.
The staining and de-staining processes require devices to hold or support the biological specimen in a way that optical imaging is possible and into which fluid reagents may be introduced or withdrawn. For example, WO2018001767A1 discloses a multilevel disposable cartridge, which is configured to accept a biological sample on a transparent support and is provided with a plurality of fluid wells or reservoirs to hold a plurality of fluid reagents.
On the other hand, most staining experiments for biological specimen like pathology examinations are performed on standard microscope inspection slides known for decades. Accordingly, many devices comprising wells or reservoirs for fluid reagents, attached to a standard microscope inspection slide have been developed. So called “chambered slides” are for example disclosed in EP0014007A1, EP0210071B1, EP1222462B1, GB2127577A, U.S. Pat. No. 3,883,398 and DE2157150. The devices disclosed herein comprise a plastic structure providing reservoirs or wells either glued or attached by mechanical means like clips or clamps to a microscope inspection slide.
However, the known chambered slides suffer from leakage and/or can not be disassembled to store the microscope inspection slide after use with the sample only. Accordingly, there is a need for chambered slides which can be easily assembled/disassembled without the danger of leaking.
Object of the invention is a slide chamber comprising a top member (10), a fluidic seal (20), a transparent closure member (30) and base member (40) wherein the top member (10) is provided with at least one (1) examination chamber (11) and a plurality of openings (12) positioned at two sides of the cover member outside of the examination chamber (11); and
the base member (40) is provided with interconnecting means (41) complementary to the openings (12) of the top member wherein
the interconnecting means (41) of the base member (40) and the openings (12) of the top member (10) are configured to mechanically interlock with each other by lateral movement thereby pressing the top member (10) against the transparent closure (30) member, preferable in a water-tight manner.
Further object of the invention is a process for preparing at least one biological tissue in a slide chamber as defined in the claims for optical examination characterized by placing the at least one biological tissue on a transparent closure member (30), combining the top member (10) and the transparent closure member (30) with an interleaved fluidic seal (20); attaching the base member (40) so that the interconnecting means (41) of the base member (40) are inserted into the openings (12) of the top member (10) and closing the slide chamber by lateral movement of the top member (10) against the transparent closure member (30), preferable in a water-tight manner.
Various exemplary details are described with reference to the following figures, wherein:
It should be understood that the drawings are not necessarily to scale, and that like numbers may refer to like features.
The slide chamber as disclosed herein is especially useful for optical examination under wet conditions, i.e. under presence of a fluid, for at least one biological tissue located inside in one or more examination chambers (11). The term “optical examination” refers to any examination method known in biology or medicine involving a biological sample and a fluid containing at least one reactant. This term includes visible light microcopy, fluorescence microscopy and staining with or without antibody conjugated dyes. Since the slide chamber according to the invention may comprise one (1) or more, like 2, 4, or even 8 examination chambers and fluids can easily introduced and removed from these, the slide chamber of the invention can be used in a multi-staining and de-staining process, for example as disclosed in U.S. Pat. No. 7,741,045 B2, EP 0810 428 B1, DE10143757, EP3037821A1, EP2725359B1 or WO2017144338A1. It should be noted that the terms “sample”, “biological tissue” or “biological sample” are used synonymous hereinafter.
After the examination is performed, the slide chamber of the invention may be disassembled, so that the sample located on the transparent closure member (30) can be retrieved and optionally stored for further examination. Accordingly, the interconnecting means (41) and the openings (12) are configured to be mechanically disengaged from each other by lateral movement of the of the base member (40) against the top member (10).
Essential for optical examination of a sample covered or at least wetted by a fluid is the water-tight assembly of the parts (members) of the slide chamber. This requires a sufficient pressure on the fluidic seal (20) combining the top member (10) and the transparent closure member (30).
To this end, the interconnecting means (41) of the base member (40) and the openings (12) of the top member (10) are preferably configured as bayonetted joint. As shown in
Preferably, the interconnecting means (41) of the base member (40) and the openings (12) are configured as to create an increasing pressure (force) between the base member (40) and the top member (10) during the closing process, i.e. during lateral movement of the of the base member (40) against the top member (10).
In a first variant, increasing pressure (force) between the base member (40) and the top member (10) is achieved by providing the interconnecting means (41) of the base member (40) and the openings (12) with a tilted shape as shown in
In a further embodiment of the invention, the increasing pressure (force) between the base member (40) top member (10) is provided by the fluidic seal (20), which is slightly compressed during the closing process, i.e. during lateral movement of the of the base member (40) against the top member (10). In this embodiment, it is not necessary to provide the interconnecting means (41) of the base member (40) or the openings (12) with a tilted shape. This embodiment is shown in
In a further embodiment of the invention, the top member (10) is provided with a recess for the fluidic seal (20). The recess for the fluidic seal (20) may be provided only around or at the outer circumference of the examination chambers. In a variant thereof, if a plurality of examination chambers is used, the recess and the fluidic seal (20) may be provided around or at the position of each examination chamber. Of course, the shape of the fluidic seal (20) determines the shape of the recess and preferable, the fluidic seal (20) seals off each examination chamber as for example shown in
The fluidic seal (20) and the top member (10) may be provided with one or more mounting means like the projection (22) and the recess/through-hole (13) as shown in
In best case, a biological tissue is placed within one examination chamber, i.e. the area of the examination chamber and the seal is larger than the sample. However, it might occur that the biological tissue is placed within at least two examination chambers, i.e. either by bad positioning of the sample or if the sample is too large for one examination chamber. In this case, the seal should nonetheless prevent leaking of fluid from one examination chamber to another and/or out of an examination chamber. In other words, the fluidic seal should not be too hard or to soft. Preferred materials for the fluidic seal (20) have a Shore A hardness of 20-50.
For easy positioning of the transparent closure member (30) i.e. of the sample against the examination chambers, the top member (10) is preferable provided with means for holding the transparent closure member (30) as shown by (14) in
To aid assembly and disassembly of the slide chamber, the top member (10) may be provided with at least one grip for example as shown with (14) in
Optical examination methods often require adding fluids containing appropriate reactants like dyes to the sample. The slide chamber of the invention is therefore provided with one or more examination chambers (as shown in
In general, “open” examination chambers are preferred. However, in a variant of the invention the examination chambers are provided with a top cover optionally having at least one orifice to allow input and removal of fluids and radiation.
The base member (40) and the top member (10) can be manufactured from PET, PS, PE, PP or COP for example by injection-moulding. Preferable, the top member (10) is transparent to visible and/or UV radiation and has a low fluorescent background, which can be achieved by using COP for manufacturing.
The slide chamber according to the invention may be used in all application of biology or medicine where cells or biological tissue is subjected to optical examination under wet conditions. This includes microscopy, in situ cultivation of cells during microscopy, life cell microscopy or ultrasonic microscopy, preferable involving a process comprising staining, incubation, washing and removal of stain on the biological sample
While various details have been described in conjunction with the exemplary implementations outlined above, various alternatives, modifications, variations, improvements, and/or substantial equivalents, whether known or that are or may be presently unforeseen, may become apparent upon reviewing the foregoing disclosure. Accordingly, the exemplary implementations set forth above, are intended to be illustrative, not limiting.
| Number | Date | Country | Kind |
|---|---|---|---|
| 19163639.8 | Mar 2019 | EP | regional |
