GB2130395A

GB2130395A - Container units for cyto-centrifuges

Info

Publication number: GB2130395A
Application number: GB08327887A
Authority: GB
Inventors: Egon Rodel; Gerhart Schroter
Original assignee: Heraeus Christ GmbH
Current assignee: Heraeus Sepatech GmbH
Priority date: 1982-11-18
Filing date: 1983-10-18
Publication date: 1984-05-31
Also published as: FR2536306A1; GB8327887D0; DE3242500A1

Abstract

Container unit for performing cyto-centrifugation, in particular for insertion into vessels such as pivoted beakers of centrifuge swing-out rotors. A chamber block 1 is held fast by clamping springs 4 in a container carrier 5 and a microscopic object carrier 6 is situated between the two. The chamber block is provided with at least two passages 2 arranged at right angles to the object carrier 6 and passing rectilinearly through the chamber block. The arrangement may have different embodiments and is easy and safe to handle. Single decanting is also possible without using filter paper. A specimen and a control may be examined at the same time. <IMAGE>

Description

SPECIFICATION Container units for cyto-centrifuges The present invention relates to container units for performing cyto-centrifugation, that is the technique of cell preparation from suspensions for medical laboratory examinations such as light or electron screen microscopy, as utilised in particular by cytological laboratories for biological fluids which contain cells. The cells settle on microscopic object carriers during the centrifugation. Their distribution, shape and size then allows of drawing conclusions regarding illnesses or other variables, if applicable. In the case of known cyto-centrifuges, the operation was conducted by the so-called filter paper technique, meaning that the fluid flows into a filter paper strip whilst doing so, whilst the cells settle on the object carrier.Difficulties arose in this connection regarding hygienic safety and the risk of cell losses.

It is an object of the invention to provide an arrangement which allow of totally renouncing filtration moreover, during sedimentation of cells orthe like.

Accordingly, the invention consists of a container unit for a cyto-centrifuge comprising a chamber block, a container carrier and a microscopic object carrier, a filter or a seal being situated between the chamber block and the object carrier, wherein the chamber block contains at least two passages passing rectilinearly therethrough and arranged at right angles or substantially at right angles to the object carrier, which passages form container cavities, and the chamber block containing the container cavities is held in operating position by spring elements arranged on the container carrier.

The advantages obtainable by means of the invention are primarily the following: After centrifuging, it is merely necessary to decant excess fluid: the costs of the filter paper are eliminated: the container unit is sealable: cell superimpositions or displacements caused by the draining of the fluid through the filter paper, are avoided.

By means of the invention, it is possible for the first time to examine twin samples simultaneously on each object carrier, such as for example a principal and a control sample of one and the same patient.

In a particularly advantageous embodiment, the container unit according to the invention is provided with conical passages as container cavities for reception of the sample (conical) and a cell enrichment is even possible in this manner by contrast to the prior art.

The chamber block is preferably transparent and as a result of this transparency, contaminations of the sample are immediately detectable and easy to remove.

The container unit is easily insertible into the container carrier. A particular direction of installation need not be adhered to, whilst doing so. The unit may advantageously and easily be inserted into a vessel such as a beaker (by means of adaptors, if appropriate), such as customary in the case of centrifuges comprising swing-out rotors. A vessel of this nature is advantageously closable in vacuum-tight manner.

An identification (for example patient's number) is advantageously incribed prior to centrifugation on the object carrier in satisfactory visible manner, thereby reducing the risk of confusion for the samples. The identification remains uncovered by the chamber block of the container unit in the normal direction of observation.

In order that the invention may be more clearly understood, reference will now be made to the accompanying drawings which show certain embodiments thereof by way of example and in which: Figure la shows the chamber block of the container unit with two transpiercing cylindrical passages parallel to each other, Figure lh shows a chamber block comprising two tapering passages, Figure 2a shows the chamber block with a microscopic object carrier inserted into a container carrier, Figure 2b shows the subject of Figure 2a in lateral elevation, Figure 3 shows the chamber block according to Figure la together with a filter and an object carrier, Figure 4 shows the container unit for insertion into the pivoted beaker as a swing-out rotor of a centrifuge dismantled into its separate parts, and Figure 5shows a swing-out rotor comprising vessels into which container units are inserted.

Referring now to the drawings, the container units according to the invention comprise substantially rectangular transparent chamber blocks comprising two transpiercing bores in each case, which are sealed at the bottom by O-rings on the object carrier and the filler openings of which are open at the top.

Container units having cylindrical passages 2 as sample reception spaces are illustrated in Figure la, and having tapering passages 3 as a sample reception space are illustrated in Figure ib, being parallel and symmetrical to each other. Whereas the cavity, for example in Figure 1a, may amount to say 1.5 cc, it may receive an apporoximately doubled sample volume in the case of Figure 1b. A cell enrichment by a factor of approximately 5 may thereby be rendered possible (at identical diameter of the sample opening). The embodiment comprising tapering passages is preferred for this reason.

The chamber block is inserted into the container carrier together with the object carrier - as shown by Figures 2a and 2b - and thus forms a potable unit. The chamber block 1 is held fast to this end by clamping springs 4 on the container carrier 5, the object carrier 6 being situated between the two. O-rings 7 consisting of a fluoro-elastomer such as obtainable under the Registered Trade Mark "Viton", which are secured in loss-proof manner in dovetail-shaped grooves and face towards the bottom of the container, are clearly apparent. Clamping springs 4 which are fastened to the container carrier 5 are so formed that they allow of easy insertion of the chamber block whilst on the other hand holding the same fast in the operating position and during the pivotal displacement in the swing-out rotor of the centrifuge.These clamping springs have wart-like curvatures or prominances 9 which hold the container unit fast in the adaptor or centrifuge beaker or other vessels into which the unit is inserted. After the block had been closed off at the top by a cover plate 8 it also is held fast by the clamping springs 4. Two small covers 10 are situated right above the passages 2 or 3 in the cover plate, which are matched in number, shape and size to the sample recesses such as passages.

The fastening of the clamping springs on the container carrier may be performed releasably by means of screws 11. The chamber block has guiding grooves 12 as shown in Figure 3, and acting to receive and secure the clamping springs 4 at either side of the block. In Figure 3 the O-rings 7 are seen in plan view, with their dovetailed grooves, as is the rotationally and mirror symmetrical embodiment of the chamber block 1, which consists of transparent plastics material (such as acrylic resin) or the like. An object carrier having an identification face 15, such as an inscription panel may be provided prominently projecting from the chamber block end at a narrow end thereof between the container carrier and the chamber block, in the same way as a filter or paper strip 14 if it is intended to work with such strips.The side of the chamber block turned away from the side provided with the O-rings, being that which is turned towards the filter paper strip, meaning that the chamber block is turned through 1800 for the "filter technique" by contrast to the decanting technique, is then apparent at the block side oppositeiy situated to the O-rings. All the sides should be as plane as smooth as possible, to ensure sealing, and the directions of flow. The advantages of providing the identification on the object carrier are obvious, because the chamber block does not cover the same, the inscription panei projecting with respect to the same, on the contrary (see Figure 2a). Risks of confusion are practically eliminated, because observation of this identification area is also possible within the centrifuge with the rotor at rest.

The chamber blocks comprising their axially parallel sample reception passages, in particular twin passages (e.g. also quadruple, sextuple, octuple etc) may easily be cleaned and disinfected or sterilised.

Contaminations are easily detectable. An aqueous solution of glutaro-di-aldehyde is appropriate for disinfecting the container units.

To improve the adhesion of the cells on the object carrier, the same way be prepared with a thin coating of glyceric albumen or glyceric gelatine.

As apparent from Figure 4, the container unit may be dismantled, assembled and inserted into a centrifuging vessel such as a pivoted beaker of a swing-out rotor. Figure 4 shows, from the right towards the left, the cover plate 8, the chamber block 1, the container carrier 5 comprising clamping springs 4, an adaptor 16 for insertion of the container unit in a vessel, preferably a pivoted beaker of a swing-out rotor as illustrated in Figure 5 and marked 18.

The steps for carrying out the cyto-centrifugation sequence are, in particular: 1. Insertion of the chamber block into the container carrier, wherein is already present a microscopic object carrier (if appropriate also a filter paper strip in between).

2. Filling matter for examination, such as a cell suspension, into the container unit, in this case into the passages into their open filler openings from above by means of a pipette.

3. Identification of the object carrier, for example with a patient's number.

4. Insertion of the container unit into the centrifuge rotor. Followed by centrifuging at g-values of between 100 and say 1000 in the plane of the object carrier, and preferably between say 200 and 800 g, and depending on rotor diameter this corresponds tog values of 500 and 3000 min-', preferably between say 850 and 2700 min-'.

5. After centrifuging, the excess fluid is decanted, for example by being drawn off by means of pipettes or poured off.

6. The chamber block is withdrawn from the container carrier together with the object carrier, the preparations are then tinted according to known methods and covered with covering glass slides as well as fixed, meaning that the cells are stabilised by coagulation of albumen structures present therein. Tinting solutions normally contain alcohol. The tinting operation occurs in known manner in baths and for a precisely determined period. In the case of the container unit according to the invention, the fixing action may be performed right after decanting of excess fluid, by means of alcohol or aldehyde. Because of the rectilinearly transpiercing passages in the chamber block in the container unit according to the invention, the fixing action by means of the fixing agent may also be performed in vapour form or aerosol form, for example from a spray can.

It is only then that the object carrier will be removed, with the cell sediment (thin layer) and that the tinting operation will be performed.

The tinting operation is normally followed by the embedment of the cell preparation in gelatine and by covering with small glass slides. It is only then that the actual microscopic examination is performed.

Applications: Cyto-centrifugation within its widest meaning is of interest for a large number of medical disciplines: Cytology Immunology Cytopathology Paediatrics Urology Gerontology Anatomy Embryology Genetics Toxicology Nuclear Medicine Forensic Medicine Neurology Biometry Hematology Oncology for biologists moreover: Botanists Virologists Zoologists Biotechnologists Marine biologists Biochemists Bacteriologists Cytochemists Paleobotanists Paleologists but also for other important ancillary subjects: Pharmacology Mineralogy Geology Crystal examinations on Sewage chemistry the object carrier Aerosol chemistry Suspended substances The special oncology (cancer research) takes pride of place in the range of application but cancer research also does so in conjunction with all other aforesaid medical research subjects (in particular urology and cytopathology).

The most important medical specimen fluids in which cells are examined, are: Liquor (cerebral fluid) Prostatic fluid Cerebrospinal fluid Tissue fluid (spinal cord puncture) Sputum (saliva) Urine Blood cells Breast fluid Lymphatic fluid Fleural puncture (costal Gallbladder fluid pleura fluid) Other possibilities of application of the cytosystem: Examinations other than optical microscopy, which require precipitates of substances in thin layers: 1. Electron screen microscopy 2. Immunofluorescence 3. Transirradiation electron microscopy 4. Autoradiography amongst others.

Claims

1. A container unit for a cyto-centrifuge comprising a chamber block, a container carrier and a microscopic object carrier, a filter or a seal being situated between the chamber block and the object carrier, wherein the chamber block contains at least two passages passing rectilinearly therethrough and arranged at right angles or substantially at right angles to the object carrier, which passages form container cavities, and the chamber block containing the container cavities is held in operating position by spring elements arranged on the container carrier.

2. A container unit as claimed in claim 1, wherein the chamber block is insertible into the container carrier with a side which is either provided with seals or turned towards a filter paper.

3. A container unit as claimed in claim 1 or 2, wherein the chamber block has cylindrical or tapering passages one side of which is turned towards the microscopic object carrier at the bottom of the container carrier, contains sealing rings whose diameters match the smallest passage diameter.

4. A container unit as claimed in claim 3, wherein the chamber block is constructed in rotational and mirror symmetry, and the side which faces away from the side containing the sealing rings and is oppositely situated with respect to the same is formed as a side turned towards a filter paper at the bottom of the container carrier.

5. A container unit as claimed in claim 3 or 4, wherein the sealing rings within the chamber block are constructed in the form of O-rings, which are secured in the chamber block in withdrawable but lossproof manner.

6. A container unit as claimed in claim 5, wherein the sealing rings consist of a fluoro-elastomer.

7. A container unit as claimed in any of the preceding claims, wherein the chamber block has guiding grooves for reception of the spring elements on the container carrier.

8. A container unit as claimed in any of the preceding claims, wherein the chamber block has a cover plate which contains covers corresponding in number, shape and size to the passages which form the container spaces.

9. A container unit as claimed in any of the preceding claims, wherein the cover plate is removable from the chamber block to allow of removal of excess fluid from the passages by being poured out or drawn off by pipettes.

10. A container unit as claimed in any of the preceding claims, wherein the chamber block consists of a transparent plastics material.

11. A container unit as claimed in claim 10, wherein the chamber block consists of acrylic resin.

12. A container unit as claimed in any of the preceding claims, wherein, if a filter paper is interposed between the chamber block and the microscopic object carrier, a liquid collector vessel at least partially surrounds the container unit.

13. A container unit as claimed in any of the preceding claims, wherein the vessel is constructed to surround the container carrier and chamber block and as a container of centrifuge swing-out rotors, into which the container unit is insertible, by means of adaptors if appropriate.

14. A container unit as claimed in claim 13, wherein the reception vessel is closable in vacuum-tight manner.

15. A container unit as claimed in any of the preceding claims, wherein the chamber block is smaller than the object carrier and the latter is provided with an identification which remains uncovered by the chamber block.

16. A container unit as claimed in any of the preceding claims, wherein the spring elements which are arranged on the container carrier, have wart-like prominances which hold the container unit fast in the vessel, in particular in the vessel of centrifuge swing-out rotors, by means of adaptors if appropriate.

17. Container units substantially as hereinbefore described with reference to the accompanying drawings.