FR2536306A1 - SAMPLE RECEIVING DEVICE FOR CENTRIFUGES USED IN CYTOLOGY - Google Patents

Abstract

THIS UNIT IS USED IN "CYTOCENTRIFUGATION", THAT IS IN THE TECHNIQUE OF OBTAINING, FROM SUSPENSIONS, CELLULAR PREPARATIONS INTENDED FOR MEDICAL LABORATORY EXAMINATIONS, SUCH AS PHOTRONOELECTRIC MICROSCOPY AND ELECTRIC MICROSCOPY A SWEEPING, AS THOSE PRACTICES IN PARTICULAR BY CYTOLOGICAL LABORATORIES ON BIOLOGICAL LIQUIDS CONTAINING CELLS. THE UNIT CONTAINS A CELL BLOCK 1, A CONTAINER HOLDER 5 AND A MICROSCOPIC OBJECT HOLDER. THE ALVEOLE 1 BLOCK CONTAINS AT LEAST TWO STRAIGHT AND CROSSING CAVITIES PERPENDICULAR TO THE OBJECT HOLDER AND INTENDED TO RECEIVE SAMPLES. THE UNIT CAN BE PLACED, POSSIBLY USING AN ADAPTER 16, IN A CONTAINER 17, IN PARTICULAR IN AN OSCILLATING CONTAINER OF A CENTRIFUGE ROTOR. THE HANDLING OF THE UNIT IS EASY AND SAFE, THE LIQUID CAN BE REMOVED EASILY, ALSO WITHOUT FILTER PAPER. THE SAMPLE AND A WITNESS MAY BE EXAMINED SIMULTANEOUSLY.

Description

The invention relates to a container unit for "cyto-

  centrifugation "> clest- say for the technique of obtaining, from suspensions, cell preparations intended for medical laboratory examinations such as photoelectric microscopy and electron microscopy & scanning as those practiced in particular by aboratory cytology ques biological liquids containing cells During centrifugation, the cells are deposited on microscopic object holders Their distribution, their shape and their size?> then allow to diagnose possibly diseases or other alterations With the known "Vcytocentrifuges", we applies the so-called filter paper technique, according to which the liquid flows in a strip of filter paper, while the cells are deposited on the holder = object This technique l 5 poses difficulties with regard to hygienic safety and

the risk of cell loss.

  The invention aims to create a method and a device which also make it possible to completely dispense with filtering.

  for the sedimentation of cellutes or similar materials.

  According to the invention, a container unit for centrifuge with cytological or cytocentri'uge eue comprising a blister block, a container holder and a microscopic object holder> with the interposition of a filter or a sealing device between the block honeycomb and the object holder, is characterized in 2 that the honeycomb block contains at least two straight and through cavities, which are perpendicular or at least substantially perpendicular to the object holder and are intended to receive samples, and that the honeycomb block is maintained in position

  working by elastic elements provided on the holder

  container The main advantages achievable thanks to the invention

tion are indicated below.

  After centrifugation, it is enough to remove the liquid which covers the cellular substance deposited on the port @ 3 Sr object The cost of filter paper is eliminated The container unit can be closed tightly The overlaps and displacements of cells as a result of V 45 flow of liquid Zi

  through the filter paper are avoided.

  Thanks to the invention, it is also possible, for the first time, to simultaneously examine double samples on an object carrier, for example a main sample and a 4 sample of u-n ee'ul contcrble. and tugme petiean C eelon a wde of realization rcxetavsntage Lx of livnin lec elveolê block "resides dao conical receptor cavities? possible in

Current technology.

  When, according to a preferred embodiment, the block

  lvoéde uiré rcipient sttransparent, d'ventuellos impure-

  very contained in the samples are: ijmediately visibleo and can be elésfclaet 1) a re-anticipate unit is easily left in place in the potnet lnetas necessary, during this ati Or L opea, to respect: aornttn riciredu It is also advantageous and easy to place it in a container such as a Gr bec (nuleetà lidu dkdapteurs) camele those usually used in cn Zrîfug'acuses with back rotors in which the containers are arranged 3 so that they are moved outward under centrifugal force It is advantageous that such a container

  May be closed in a vacuum-tight manner.

  A mark (a patient number for example) is affixed in a clearly visible manner on the object-holder before centrifugation, which reduces the risk of reversals of the effects. This marking is not covered by the honeycomb block of the unit

  container in normal viewing direction.

  Other features and advantages of the invention

  will emerge more clearly from the description which follows

  of execution of non-limiting embodiment Cs, as well as of the appended drawings, in which the figure shows it in vertical section and in plan a block av representing two parallel cylindrical cavities; Figure lb is a similar representation of a honeycomb block having two conical cavities; Figure 2a is a side view partially in section of a honeycomb block like that of Figure la after its establishment, together with a microscopic object holder, in a container holder; Figure 2b is an end view of the container unit of Figure 2a; Figure 3 is a perspective view of a block

  dimpled like that of figure la with a filter and a throat-

  object; Figure 4 is a perspective view of the various components of a container unit intended to be placed in a beaker designed to be mounted oscillating in the rotor of a centrifuge where the beakers are moved outwards. centrifugal force; and Figure 5 is a perspective view of such a centrifuge rotor with containers (beakers) in which

  container units according to the invention have been installed.

  The container units according to the invention each comprise a honeycomb block 1 which is transparent and essentially has a parallelepiped shape, in which at least two through cavities are formed, which are sealed at the bottom relative to the object holder, by O-rings. for example and whose sample introduction openings, provided in

high, are open.

  Figure la shows a honeycomb block 1 which has cylindrical cavities 2 for receiving samples = The cavities 2 are parallel and are arranged symmetrically by

  relative to a median transverse plane of the alveolar block 1 The fi-

  gure lb represents a honeycomb block 1 with sample receiving cavities 3 which are conical These cavities are also arranged symmetrically with respect to a median transverse plane of the block While a cylindrical cavity 2 like those visible in the figure la can have a capacity of about 1.5 ml for example, the cavities 3 of the example shown in Figure lb can receive samples whose volume is approximately

  double It is thus possible (with the same diameter of the

  sample thickness) to obtain an enrichment in cells of a factor of approximately 5 The execution with conical cavities is therefore preferred. The honeycomb block 1 is placed, together with the object holder, in the container holder 5, as shown in the

  Figures 2 a and 2 b, so that a transportable unit is obtained.

  The block 1 is immobilized on the container holder 5 by res-

  blocking spells 4, with arrangement of the object holder 6 between the block 1 and the container holder 5 The drawings clearly show the O-rings 7, made of a fluorinated elastomer like that known under the brand "Viton" On the FIGS. 1a, 1b, 2a and 2b, the O-rings 7, held in dovetail grooves,

  are located at the bottom, i.e. facing towards the bottom of the

  pient The locking springs 4, fixed on the container holder 5, are shaped so as to allow an easy installation of the honeycomb block and nevertheless to ensure a solid holding of the block in the working position and during oscillation and rotation in the centrifuge The blocking springs have bulges 9 for holding the container unit in place in an adapter, u Beaker of centrifuge or in another container intended to receive the unit Block 1 has previously been covered at the top of a cover plate 8, which is also held by the locking springs 4 This cover plate has small covers 10 which are located directly above the cavities 2 or 3 and the number, shape and the size are adapted to those of the sample receiving cavities The locking springs can be removably fixed to the container holder by screws 11 The honeycomb block 1 has guide grooves 12, visible s ur in Figure 3, which are intended to receive and hold in place the locking springs 14 on both sides of the block, In the representation of Figure 3, the O-rings 7, held in place in the tail grooves dovetail, are located at the top This figure also shows the symmetrical embodiment of the honeycomb block 1 with respect to a central plane As already mentioned, the block will generally be made of a transparent material, of acrylic glass for example, Between the container holder and the honeycomb block can be arranged an object holder provided with a marking area 15, with an inscription area for example, which is located on a

  short side of the object holder 6 and which protrudes from the cor-

  corresponding to the honeycomb block A strip of filter paper 14 of similar shape can be placed between the object holder 6 and the

  honeycomb block 1 in case it is necessary to work with such strips.

  In such a case, that is to say if it is necessary to work according to the filtering technique and not according to the decantation technique, the honeycomb block 1 is turned over (turned 180), so that the toroidal jnints 7 are located at the top The opposite faces of the block must be as flat and smooth as possible to ensure watertightness and 12 suitable flows The marking of the object holder represents an obvious advantage, especially since it is not covered by the honeycomb block (we can clearly see in Figure 3 that the marking range exceeds block 1) o The risks of reversals are thus practically excluded, the marking also remaining visible

  in the centrifuge when the rotor is stopped.

  The honeycombed blocks with their cavities for receiving samples with parallel axes can be easily cleaned and disinfected or sterilized Stains are easily visible The disinfection of the container units can be done with an aqueous solution of glutaric dialddhyde The blocks have in particular two cavities but it is perfectly possible to provide 4, 6, 8 or any number of cavities

in each block.

  In order to improve the adhesion of the cells on the holder

  object, it can be coated beforehand with a thin layer

  egg white glycerin or gelatin glycerin.

  As can be seen in Figure 4, a container unit according to the invention can be decomposed and assembled to be placed in a centrifuge container such as an oscillating beaker. Figure 4 shows, from right to left, the plate cover 8, the honeycomb block 1, the container holder 5 2536306 i with the locking springs 4 and an adapter 16 for setting

  place the container unit in a container 17, consisting of pre-

  reference by an oscillating beaker of a rotor, like the rotor 18 shown in FIG. 5, where the beakers are mounted so as to oscillate outwards under the effect of centrifugal force

during rotation.

  If a filter paper is interposed between the honeycomb block and the microscopic object holder, a liquid collecting container can be provided which at least partially surrounds

the container unit.

  The different operations of a-eytocentrifugation are as follows 1 Placement of the honeycomb block in the container holder, where

  already finds a door = microscopic object (possibly covered

  green of a strip of filter paper) 0 2 Introduction of the material & examine such as a cell suspension in the container unit, i.e. in the cavities, by means of a pipette and through the openings at the top

two cavities.

  3 Marking of the slide, for example with a patient number

  4 Positioning of the container unit in the centrifuge rotor

  fugeuseo The consecutive centrifugation is carried out with numbers g, in the plane of the object-holder, between 100 and approximately 1000, preferably Erence between approximately 200 and 800 g; depending on the diameter of the rotor: this corresponds to g numbers of 500 and 3,000 min 1 l

  preferably between around 850 and 2700 min -

  After centrifugation, remove the liquid that covers the

  deposit on the object holder, for example by pipette or by pouring

sement.

  6 The honeycomb block with the object holder are removed from the holder

  container and the preparations are then stained according to known methods, covered with covering glass and fixed in the sense that the cells with their albuminum structures are stabilized by coagulation The staining solutions normally contain glue The staining is carried out in a known manner in baths and for a fixed time with

2536306 '

  precision With the container unit according to the invention, the fixing can be carried out with alcohol or aldehyde immediately after the removal of the liquid covering the deposit. In raisondas straight through cavities of the honeycomb block in a container unit according to the the invention can also be carried out using a fixing agent in the form of a vapor or aerosol,

  for example from a spray bottle.

  It is only then that we remove the object with the deposit or cellular sediment (thin layer) and that we

does the coloring.

  The coloring is normally followed by the coating of the cell preparation in gelatin and its covering with glass plates The microscopic examination properly

said only takes place afterwards.

  Applications Cytocentrifugation in its broadest sense is interesting for a large number of medical disciplines: cytology immunology

pediatric cytopathology-

  urology gerontology anatomy embryology genetic toxicology nuclear medicine general medicine neurology biometry hematology oncology and also in biology for: botanists virologists zoologists biotechnologists marine biologists biochemists bacteriologists cytochemists paleobotanists paleologists but also for other important bordering fields pharmacology mineralogy structural examinations

crystal geology on the carrier

  object wastewater chemistry aerosol chemistry suspended matter

aerosol chemistry-

  At the very top of the range of applications is

  finds especially oncology (oncology) but also can-

  cerology in cooperation with all other disciplines of

  medical research mentioned above (especially uro-

logy and cytopathology).

  What matters most to cytologists during examinations are qualitative phenotypes in the sediment

  cellular, i.e., for example, morpho-

  logic of cells or cell nuclei, or the appearance of

  types of cells that had not been planned, and so on.

  At most, quantitative assessments relate to

  the relationship of different types of cells to each other.

  The most important organic sample liquids in which cells are examined are the following cerebral fluid prostatic fluid cerebrospinal fluid tissue fluid (fluid obtained by saliva puncture the spine) urine blood cells breast fluid lymphatic fluid liquid obtained by fluid puncture pancreatic pleurocentesis Other possibilities of application of the system described here are examinations other than by ordinary microscopy (optical) which require costs or precipitations of substances in thin layers, such as: 1 scanning electron microscopy 2 imunofluorescence 3 microscopy electronic transmission 4 autoradiography

and even more.

9-

Claims (14)

  1 container unit for cytological centrifuges or   cytocentrifuges, comprising a honeycomb block (1), a holder   container (5) and a microscopic object holder (6), with inter-   position of a filter M 4) or of a sealing device (7) between the honeycomb block and the object holder, characterized in that the block   honeycomb (1) has at least two straight and transverse cavities   santes (2, 3), which are perpendicular or to the moias substantially perpendicular to the object holder (6) and are intended to receive samples, and that the honeycomb block (1) is held in the working position by elastic elements (4) provided on the container holder (5).   2 unit according to claim 1 i characterized in that the honeycomb block (1) can be arranged in the container holder (5) by a face carrying seals (7) or by a face   intended to be applied against filter paper (14).   3 Unit according to claim 1 or 2, -characterized in that the honeycomb block (1) has cylindrical cavities (2)   or conical (3), one end of which, that facing towards the holder   microscopic object (6) on the bottom of the container holder (5), is provided with a sealing ring (7) whose diameter is adapted   to the smallest diameter of the cavity.   4 Unit according to any one of the preceding claims.   cédentes, characterized in that the honeycomb block (1) is produced symmetrically with respect to a central plane and in that 1 Xa face opposite to that containing the sealing rings (7> is designed to be applied against a paper filter (14)   arranged on the bottom of the container holder (5).   Unit according to any one of the preceding claims.   cédentes, characterized in that the sealing rings provided in the honeycomb block (1) are O-rings (7) which are   detachably held in the block.   6 unit according to claim 5, characterized in that   that the O-rings (7) are made of fluorinated elastomer. 2536306 '   7 Unit according to any one of the preceding claims.   teeth, characterized in that the honeycomb block (1) has   guide grooves (12) for receiving the elastic elements.   ticks (4) provided on the container holder (5).   8 Unit according to any one of the preceding claims.   denges, characterized in that the honeycomb block (1) has a cover plate (8) which contains GO covers) whose number, shape and size correspond to those of the cavities   sample receivers (2, 3).   IO 9 Unit according to any one of the preceding claims.   teeth, characterized in that the cover plate (8) can be removed from the honeycomb block (1) to remove cavities (2, 3), by pouring or by pipette, from the liquid   covering the deposits formed on the object holder (6).   10 Unit according to any one of the preceding claims.   teeth, characterized in that the honeycomb block is made of a   transparent synthetic material, in particular acrylic glass.   1 l Unit according to any one of the preceding claims.   teeth, characterized in that, if a paper is inserted   filter (14) between the honeycomb block (1) and the micros-   copic (6), a liquid collecting container surrounds at least partially the container unit.   12 Unit according to any one of the preceding claims.   teeth, characterized in that it is combined with u Ln container which surrounds the container unit with the honeycomb block (1) and which consists of a container (17) of a centrifuge rotor whose containers are movable towards the under the effect of centrifugal force, the container unit possibly being   be placed in this container using an adapter (16).   13 Unit according to claim 12, characterized in that the receiving container (17) can be sealed empty.   14 Unit according to any one of the preceding claims.   teeth, characterized in that the honeycomb block (1) is smaller than the object holder (6) and in that the latter is likely to be provided with a marking (15) which is not covered by the honeycomb block. 25363 o 6 '   Unit according to any one of the preceding claims.   cédentes, characterized Ae in that the elastic elements (4), provided on the container holder (5), have bulges_ (9)   to keep the unit in place in a container,   using an adapter (16), in particular in a container (17) of a centrifuge rotor carrying containers   movable inward under the effect of centrifugal force.