DE8232257U1 - VESSEL UNIT FOR CYTOCENTRIFUGES - Google Patents

Description

Hanau, November 15, 19SZ ZPL-Zw / ha

Heraeus Christ GmbH, Osterrods

"Vessel unit for cytocentrifuges"

The invention relates to a vessel unit for performing cytocentrifugation, that is to say the technique of cell preparation from suspensions for medical laboratory tests such as light and scanning electron microscopy, like them used in particular by cytological laboratories for biological fluids containing cells. at After centrifugation, the cells settle on microscopic slides. Their distribution, shape and size leaves then, if necessary, for diseases or other changes conclude. In known cytocentrifuges, the so-called filter paper technique was used, i.e. the liquid flows into a strip of filter paper while the cells are deposited on the slide. Passed it Difficulties in terms of hygienic safety and the risk of cell loss. Object of the present invention is to create a method and an apparatus that also allows filtration during sedimentation to completely dispense with cells or the like. This object is achieved by the features listed in claim 1 Further refinements of the invention are set out in the subclaims and the description and drawings of exemplary embodiments refer to. The invention also includes all combinations and sub-combinations of those shown and claimed features described both with each other and with features known per se.

The advantages that are achievable with the invention are especially the following:

After centrifuging only supernatant is needed decanted to «/ earth. The cost of the filter paper is eliminated. The vessel unit can be sealed. Cell overlays or displacements as a result of the liquid flowing out through the filter paper M / earth avoided.

With the invention it is possible for the first time to carry out duplicate samples such as main and control samples on each slide examine one and the same patient at the same time. The vessel unit according to the invention is special in one aspect advantageous embodiment provided with conical channels as vessel cavities for receiving the sample (conical) and in this way, in contrast to the prior art, even cell enrichment is possible.

As a result of the transparency of the chamber block of the vessel unit, contamination of the sample can be recognized immediately and easy to remove.

The tube unit can easily be inserted into the tube carrier. It is not necessary to pay attention to a particular installation direction. The unit is also in a container like mug It is advantageous to use it easily (if necessary by means of adapters), as is customary in centrifuges with swing-bucket rotors are. Such a container can advantageously be closed in a vacuum-tight manner.

An identification (e.g. patient number) is clearly visible on the slide before centrifugation attached and thereby reduces the risk of confusion for the samples. The marking remains in normal view

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Direction of the chamber block of the vessel unit uncovered. Further Advantages result from the description below From exemplary embodiments.

Show it

Figure la the chamber block of the vessel units with two continuous cylindrical channels parallel to each other,

Figure Ib shows a chamber block with two conical channels (conical),

Figure 2a the chamber block with microscopic slide inserted into a vessel holder,

Figure 2b shows the subject of Figure 2a in a side view ,

Figure 3 the Kammefblock according to Figure Ip together with a filter and a slide,

Figure 4 shows the vessel unit to be inserted into the Schv / enlbecher dismantled into its individual parts as a cut-out of a centrifuge,

Figure 5 a cut-out with ": Containers in which Cargo units are networked,

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The vessel units according to the invention consist essentially of cuboid formines through the i chtirjen comb lure with each

two through holes through O-rings on the bottom sealed on the slide and their filling openings are open at the top.

In Figure la are vessel units with cylindrical channels 2 as a rehearsal space and in figure lb with conical (Conical) channels 3 shown as a sample receiving space parallel and symmetrical to each other. During the For example, the cavity in Figure la can be about 1.5 ml, it can be about twice as large in Figure Ib Take up the sample volume. This means that cells can be enriched by etu / a (with the same diameter of the sample opening) Factor 5 can be made possible. The design with conical channels is therefore preferred.

The Ksmmerblock comes together with the slide - like Figures 2a and 2b show - inserted into the vessel carrier and thus forms a transportable unit. The chamber block 1 is held by clamping springs 4 on the vessel carrier 5, wherein the slide 6 is arranged between the two. The O-rings 7 are clearly visible from one Fluoroelastomer such as is available under the trade name "Viton", which is held captive in dovetail grooves and face the bottom of the vessel. The clamping springs 4, which are attached to the vessel carrier 5, are like this shaped so that they allow easy insertion of the chamber block, on the other hand this in the working position and Hold the centrifuge's swing-bucket rotor during the swivel movement. These spring clips have bulbous bulges or elevations 9, which hold the vessel unit in the adapter or centrifuge ugeribecher or other containers in which the unit is inserted. After the block above covered with a cover plate Q, which is also covered by the clamping springs 4 is held in place in the cover plate are two small lids 10 directly above the channels 2 relational way 3 arranged in counting, form

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and size are adapted to the Prabeausnehmungen and / or channels. The fastening of the clamping springs can be detachable by means of screws 11 take place on the vessel carrier. The chamber block has guide grooves 12, as can be seen in Figure 3 and for receiving and holding the clamping springs 4 on both Serving sides of the block. In Figure 3, the O-rings and their dovetail grooves can be seen from above, likewise u / ie the centrally symmetrical and mirror-image execution of the chamber block 1, which entern from transpar Plastic, such as acrylic glass) or the like. A slide can be placed between the vessel carrier and the chamber block with identification field 15, such as writing field, are also provided on a narrow end protruding from the chamber block end, like a filter or paper strip 14, if with such Strip to be worked. The side of the chamber block facing away from the side provided with O-rings can then be seen, the one facing the filter paper strip is, that is, the chamber block is used for the "filter technology" In contrast to decanting technology, rotated 180 ° to the side of the block opposite the O-rings. All sides should be as flat and smooth as possible in order to ensure the seal or To ensure flow directions. The advantages of placing the marking on the slide are obvious, because the chamber block does not cover this, but the text field protrudes from it (cf. Figure 2a). There is practically no risk of confusion because the view is up this identification field also in the centrifuge when it is at a standstill Rotor is possible.

The chamber blocks with their parallel-axis sample receiving channels Double channels in particular (e.g. also four, six, eight, etc.) can be easily cleaned and disinfected or sterilize, soiling is easily recognizable. A is suitable for disinfecting the vessel measuring units aqueous solution of giutartdialdehyde.

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To improve the adhesion of the cells to the slide, this can be prepared with a thin layer of glycerine protein or glycerine gelatin.

As can be seen from Figure 4, the vessel unit can be dismantled, assembled and inserted into a centrifuge container like swivel bucket a swing-bucket rotor. Figure 4 shows the cover plate 8 from right to left, the Kanrnrerblock 1, the vessel carrier 5 with clamping springs 4, one Adapter 16 for inserting the vessel unit into a container 17, preferably a swivel cup of a swing-bucket rotor, as shown in Figure 5 and labeled 18.

The steps in performing cytocentrifugation are in detail:

1. Insertion of the chamber block into the tube carrier, which already contains a microscopic slide (if necessary there is also a strip of filter paper in between).

Z. Filling in test material, such as cell suspension in the vessel unit, here in the channels using a pipette from above into their open filling openings.

3. Marking the slide, for example with a Patient number.

4. Insertion of the tube assembly into the rotor of the centrifuge. Then centrifuge with cj-Zahlon in the plane of the slide between 100 and about 1000, preferably between etwn 200 and 800 g and depending on the rotor diameter diets y numbers of 500 and "JOOOrnin", preferred zuiüchtiri

about 850 and 2700min

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5. After centrifugation, the supernatant liquid decanted, for example, by pipetting or pouring.

6. The chamber block is removed from the vessel carrier together with the slide, the preparations are then colored according to known methods and covered with cover glasses and fixed, that is, the cells and their existing protein structures are stabilized by coagulation. Usually staining solutions contain alcohol. The dyeing is done in a known manner ^ n bathrooms and a well-defined time. In the case of the vessel unit according to the invention, fixation can take place immediately after the supernatant liquid has been decanted by means of alcohol or aldehyde. As a result of the straight through channels in the container unit according to the invention in the Kgmmerblock, the fixation with the fixative can also take place in vapor form or as an aerosol, for example from a spray bottle

Only then will the slide with the cell sediment (thin layer) be removed and the staining carried out.

The staining is usually followed by embedding the cell preparation in gelatin and covering it with small glass slides at. Only then is the actual microscopic examination carried out.

Applications:

Cytocentrifugation in the broadest sense is interesting for a large number of medical disciplines:

cytology
Cytopathology
urology
anatomy

Immunology Pediatrics Gerontology Embrylogic

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genetics

Nuclear Medicine Neurology Maematology

Toxicology Forensic Medicine Biometry Oncolgy

also for biologists:

Botanist zoologist

Army biologists, bacteriologists, paleobotanists

Virologists

Biotechnologists

biochemist

Cytochemist

Paleologists

but also for other important border areas:

Pharmacology mineralogy geology
Wastewater chemistry

Aerosol chemistry

Crystal examinations on the slide

Schvi / ebsto f f e

At the top of the application area is special oncology (cancer research) but also cancer research in cooperation with all other medical research institutions mentioned above (especially urology and cytopathology).

The cell researchers are primarily interested in the qualitative appearance in the cell sediment that means, for example, morphological changes in cells or cell nuclei or the appearance of cell types, that were not expected, etc.

Quantitative evaluations relate at most to the ratio of different cell types to one another.

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pie u / ichtsten (lisdical sample liquids in which Cells to be examined:

(.Cerebral fluid) cerebrospinal fluid (Spinal cord puncture) urine

Breast fluid pleural puncture (pleural fluid)

Prostate fluid Tissue fluid Sputum (saliva) Blood cells
Lymph fluid

Bile

Other possible uses of the cytosis system:

Investigations other than light microscopy, the substance precipitations in thin layers require:

1. Scanning electron microscopy

2. Immunofluorescence

3. Transmission electron microscopy

4t autoradiography a.o.

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Claims (15)

■ i · I · ■ · t ■ * it I} 1 · » t t I I * Hanau, November 15, 1982 ZPL-Zvu / ha Heraeus Christ GmbH, Osterrode "Vessel unit for cytocentrifuges" Expectations Vessel unit for cytocentrifuges with one chamber block, a tube holder and a microscopic slide, with a filter between the chamber block and slide or a seal is provided, characterized in that the Karr, lerblock at least two perpendicular or Contains straight through channels arranged essentially perpendicular to the slide, the vessel cavities form and the chamber block containing the vessel cavities is held in the working position by arranged on the vessel carrier spring elements. 2. Vessel unit according to claim 1, characterized in that the chamber block either with one with seals provided or a filter paper facing side can be used in the vessel carrier. 3. Vessel unit according to claim 1 or 2, characterized in that the chamber block has cylindrical or conical channels has, one side of which faces the microscopic slide at the bottom of the vessel carrier, sealing rings contains the diameter of which is adapted to the smallest duct diameter, Vessel unit according to one of the preceding claims, characterized in that the chamber block is designed to be centrally symmetrical and mirror-inverted and the side which is turned away from the side containing the sealing rings and is opposite to it, as a filter paper on the floor the side facing the vessel carrier formed 5. Vessel unit according to one of the preceding claims, characterized in that the sealing rings in the iiammerblock designed in the form of G-rings which are removable but captive in the chamber block are. 6. vessel unit according to claim 5, characterized in that the sealing rings consist of a fluoroelastomer. 7. Vessel unit according to one of the preceding claims, characterized in that the chamber block guide grooves for receiving the spring elements on the vessel carrier. 8. Vessel unit according to one of the preceding claims, characterized in that the chamber block has a cover plate has, the number, shape and size of the channels ,, which form the vascular spaces, contains corresponding covers. 9. Gef äßeirfhei t according to one of the preceding claims ^ characterized in that the cover plate is removable from the chamber block in order to remove excess liquid by pouring or pipetting out of the channels. 10. vessel unit according to one of the preceding claims, characterized in that the chamber block is made of transparent Plastic, in particular acrylic glass, is made. tt t * " I < s · I". it till « * ΙΛ it t H i 11 »Vessel unit according to one of the preceding claims, characterized by j that between Kämmerblock And a filter paper placed on a microscope slide and a liquid collector at least partially surrounds the vessel unit. 12. vessel unit according to one of the preceding claims, characterized in that the container surrounds the vessel carrier and the chamber block and acts as a container for the centrifuge swing-out furnace is designed, in which the vessel unit can optionally be inserted by means of adapters is. 13. Vessel unit according to claim 12, characterized in that that the receptacle can be closed in a vacuum-tight manner. 14. Vessel unit according to one of the preceding claims, characterized in that the chamber block is smaller than the slide is designed and the latter can be provided with a label that is not covered by the chamber block remain. 15. Vessel unit according to one of the preceding claims, characterized in that the spring elements on the vessel carrier are arranged, have wart-like elevations, which hold the vessel unit in the container, especially in the container of centrifuge swing-out rotors if necessary by means of an adapter.