JP3207404U - Instruments for preparing cell smears - Google Patents

Abstract

An object of the present invention is to provide an adapter for preparing a smear sample and a chamber holder for centrifuging a chamber system in a centrifuge equipped with a general-purpose swing rotor. An adapter 1 has a shape that matches the shape of a bucket of a general-purpose centrifuge, and has chamber system receiving portions 2a, 2b, and 2c for holding the chamber system in the center portion. According to the shape of the bottom surface, the shape viewed from the top surface is a substantially hexagonal shape, a substantially rectangular shape, or the like. Further, when the bucket is deep, the adapter 1 is provided with a little play at the corner so as to be easily taken out, or the handle 3 is attached to the left or right or the diameter direction of the rotor. [Selection] Figure 1

Description

  The present invention relates to a device for producing a single-layer smear of cells directly on a slide glass from a sample in which suspended cells such as body fluids and cultured cells are suspended. In particular, the present invention relates to an instrument for preparing a smear without requiring a special centrifuge.

  There are two methods for creating smears in body fluids such as blood and cerebrospinal fluid and cells that are suspended like cultured cells: artificially smearing and smearing using a machine It is divided roughly into. Artificial smearing methods include the drawing glass method, the laminating method, and the natural sedimentation method, all of which require skill. For this reason, when a technician who is immature in the technique prepares a smear by an artificial method, a difference occurs in the completion of the cell smear. In some cases, the result determines the cell determination result, and an accurate diagnosis may not be made.

  On the other hand, the mechanical smearing method also called a spin method is a method of smearing cells by centrifugation. The mechanical smearing method includes an auto smear (registered trademark) method and a cytospin method. Both of these methods require dedicated centrifuges and instruments, but if the number of cells contained in the specimen is kept properly, no special procedure is required and anyone can make similar specimens. (Non-Patent Document 1).

  In hospitals, smear specimens such as cerebrospinal fluid specimens and blood specimens may be prepared to make an appropriate diagnosis. In particular, the cerebrospinal fluid test is a very important test in the pathophysiology of cerebrospinal diseases and the differentiation and observation of treatment of neoplastic diseases. In particular, the calculation and classification of the number of cells is extremely important for accuracy and speed, and high-accuracy testing is required. The Japan Society for Clinical Laboratory Engineers recommends that a cell smear be prepared and observed in detail if the number of cells is difficult to classify or if unknown (difficult to classify) cells are observed. (Non-Patent Document 2).

  Since cerebrospinal fluid specimens have a small number of cells and cell degeneration is fast, it is desirable to prepare a smear within 1 hour after collection, and an environment in which a smear can be prepared simultaneously with specimen processing is optimal. In facilities where cerebrospinal fluid examination is an emergency examination item such as overtime or on holidays, it is desirable that an unaccustomed person on duty can prepare an appropriate specimen (Non-patent Document 3).

  Artificial smear preparation methods for cerebrospinal fluid specimens include the ordinary drag glass method (ordinary method), the drag glass method using the capillary phenomenon recommended in Non-Patent Document 2, and the natural sedimentation method of Sayk. Etc. are used.

  The pulling glass method is a method in which a sample is extended to one side of a cover glass on a slide glass, and then spread on the slide glass at a constant speed by the cover glass. In the drag glass method (ordinary method), cells often gather at the end of pulling, and in a cerebrospinal fluid sample with a small number of cells, the distribution of leukocytes is not uniform and the cell distribution is often not observed. In addition, many cells are destroyed. Furthermore, skill is required because it is necessary to adjust the angle between the slide glass and the ground glass, the drawing speed of the glass, and the like depending on the viscosity of the specimen. Usually, training with cerebrospinal fluid specimens requires about 1 day to 1 week, but cerebrospinal fluid specimens are rarely submitted and the amount of specimens is small, so training is very much Have difficulty.

  In the pulling glass method using capillary action, the specimen is centrifuged and the supernatant is removed, and then the AB type serum that has been sucked into the Pasteur pipette is discharged and mixed with the sample. Next, the cells are collected by a Pasteur pipette, and a sample is prepared from the collected cells by a normal drag glass method. At this time, when the number of cells is small, there is a problem that whether the cells can be collected or not visually confirmed when the cells are collected by a Pasteur pipette. Further, since the AB type serum is mixed with the sample in order to prevent cell degeneration, the AB type serum is required for preparing a smear. Although this method is suitable for cerebrospinal fluid specimens, it is a method that applies the drag glass method, so that training is necessary as in the normal method.

  Syak's natural sedimentation method is a method in which a sample is sedimented and smeared without centrifugation. First, filter paper with a hole is placed on a slide glass, and a plastic tube having the same inner diameter as the hole is set up to match the hole. Appropriate pressure is applied to the filter paper from above, and about 1 ml of cerebrospinal fluid is injected. In this method, the water of cerebrospinal fluid is absorbed by the filter paper, and the cells are settled and smeared in the hole portion on the slide glass. This method not only requires 20-30 minutes for sedimentation, but also has the problem that cell degeneration occurs during that time. Further, the cerebrospinal fluid sample with a low cell collection rate of about 25% and a small number of cells has a problem that the number of cells to be observed is small.

  In contrast, mechanical smearing does not require training and anyone can produce a clean smear, and has the advantage that cell degeneration is less and the cells can be smeared evenly compared to the drag glass method. Furthermore, a smear can be prepared even with a sample having a small number of cells or a small sample of about 100 μL because of the cell collection method. Furthermore, the time required for specimen preparation is as short as about 5 minutes.

  However, the centrifuge used for the auto smear method and the cytospin method is a special dedicated machine and cannot be used for other purposes, so it is not so popular as it is provided in all hospital examination rooms. On the other hand, a general-purpose centrifuge equipped with a swing rotor used for urine sediment or the like is usually installed in a hospital laboratory. However, the container containing the cell suspension used in the auto smear method and the cytospin method and the chamber in which the slide glass is set have a special shape, so it is not suitable for a bucket of a general-purpose centrifuge. Therefore, a sample cannot be prepared using a general swing rotor general-purpose centrifuge.

  So far, adapters have been developed for holding test tubes suitable for swing rotors and angle rotors when test tubes having various outer diameters and bottom shapes are placed in a centrifuge (Patent Documents 1 and 2). . An adapter for separating a sample from a microplate or a specially shaped container using a general-purpose centrifuge has also been proposed (Patent Documents 3 and 4).

Tadao Kobayashi et al. Jpn. Soc. Clin. Cytol. Vol. 30 (4), p. 703-709. JAMT Technical Textbook Series Cerebrospinal Fluid Test Technology Textbook, 2015, Japan Society for Clinical Hygiene Laboratory Engineers Masahiro Ishiyama, 2012, Medical Technology Vol. 40, no. 4, p. 369-376

JP 2004-261699 A JP-T-2015-521554 JP 2003-154292 A Japanese Patent Laying-Open No. 2015-057287

  However, adapters and instruments for preparing smears using a centrifuge that is always installed in each hospital laboratory have not been developed. This invention makes it a subject to provide the adapter for producing a smear sample with the general purpose centrifuge provided with the swing rotor. The present invention also relates to a chamber holder that holds the chamber system.

  The present invention relates to an adapter for preparing a smear with a centrifuge equipped with a general-purpose swing rotor in a hospital laboratory or the like, and a chamber holder for centrifuging a chamber system. Moreover, it produces by centrifuging the adapter of any one of following (1)-(4), or the chamber holder of (5) or (6) using a predetermined centrifuge chamber or a centrifuge. It is an object of the present invention to disclose a smear preparation method characterized by this.

(1) An adapter for a bucket of a swing rotor of a centrifuge, which is provided with a chamber system receiving portion for engaging a chamber system for producing a smear for cytodiagnosis.
(2) The adapter according to (1), wherein the adapter is provided with a marker for distinguishing between an outer side and an inner side of the swing rotor in the axial direction.
(3) The adapter according to (1) or (2), wherein the adapter is provided with a handle for attaching to and detaching from the bucket.
(4) The adapter according to claim 1 or 2, wherein the adapter includes a basic unit including the chamber system receiving portion and an adjustment unit fitted to the basic unit.
(5) A chamber holder for installing a chamber system for preparing a smear for cytodiagnosis on a swing type centrifuge, and a bridging portion for bridging the swing rotor;
A chamber holder provided with a chamber system installation part.
(6) The chamber holder according to (5), wherein the weight of the portion located inside the swing shaft when stationary is greater than the portion located outside.

  Since a smear can be prepared by a spin method using a general-purpose centrifuge usually installed in any hospital, a stable cell smear can be prepared regardless of the procedure.

The perspective view of the adapter of the 1st Embodiment of this invention and the state which has arrange | positioned the chamber system are shown. The figure which shows the state which installed the chamber system in the general purpose centrifuge using the adapter of 1st Embodiment of this invention. The figure which shows the state which has arrange | positioned the chamber system to the adapter of the 2nd Embodiment of this invention, and an adapter assembly example. It is the perspective view which shows the chamber holder which is the 3rd Embodiment of this invention, swing rotor bridge | crosslinking part, and sectional drawing of an adjustment member. It is a figure which shows the state which installed the chamber system in the general purpose centrifuge using the chamber holder of the 3rd Embodiment of this invention. It is a perspective view which shows the state which installed the chamber holder in the chamber holder of the 4th Embodiment of this invention, and this chamber holder. Giemsa-stained images of smears prepared using the adapter of the present invention and smears prepared using auto smear.

  In the spin method, a clean smear can be produced by applying a centrifugal force perpendicular to the slide glass using a swing rotor. When preparing a smear using the instrument of the present invention, any swing rotor can be used. Further, since the purpose is to prepare a smear, the centrifuge need only be capable of being centrifuged at about 500 to 2000 rpm, and need not be particularly capable of being centrifuged at a high speed.

  Hereinafter, the container for cell suspension used in the auto smear method and cytospin method, and the centrifuge tube for cell collection are referred to as a chamber, combined with a slide glass, filter paper, etc., and set so that a smear can be prepared by centrifugation. This is called a chamber system.

  Since the adapter of the present invention is used for preparing a smear for cytodiagnosis, it is used in a relatively low-speed centrifugation. Therefore, any material can be used as long as it can withstand centrifugation at about 500 to 2000 rpm. Examples of such materials include polypropylene resins, polyethylene resins, styrene resins, acrylic resins, polyamide resins, polyester resins, and other synthetic resins, nitrile rubber, acrylic rubber, polyisoprene rubber, and other rubbers. Can be mentioned. Further, when used in a hospital, patient specimens are handled, so there is a possibility of contamination with bacteria, viruses, and the like. Therefore, since it is necessary to wash and sterilize when contaminated, a material that can withstand washing and sterilization is preferable. As a cleaning method, chlorine-based and alkaline detergents are often used, and therefore, a method that can withstand the use of these detergents is preferable. Moreover, it is preferable that it is a material which can endure high temperature and high pressure like autoclave sterilization.

  In addition, the chamber holder of the present invention can be manufactured from a material generally used as a bucket of a centrifuge, such as metal, for example, aluminum, titanium, and an alloy containing these metals.

  Hereinafter, the adapter and chamber holder adapted to the shape of a commercially available chamber system mainly used in the auto smear method will be described. However, it goes without saying that if the shape of the portion of the adapter with which the chamber system is engaged is replaced with the shape of the chamber system used in the cytospin method, it can be used in the cytospin method.

[Example 1] Adapter using an existing bucket For a general-purpose swing rotor, a bucket for holding a centrifuge tube is prepared in several shapes and sizes according to the size and number of centrifuge tubes. Yes. The adapter 1 of the present invention has a shape that matches the shape of the bucket of a general-purpose centrifuge, and may have chamber system receiving portions 2a, 2b, and 2c for holding the chamber system at the center (FIG. 1). FIGS. 1A to 1D show adapters shaped to fit a commercial chamber system used in the auto smear method, and FIGS. 1E and 1F show adapters shaped to fit a cytospin sample chamber.

  For example, as illustrated in FIG. 1, the adapter 1 of the present invention has a substantially hexagonal shape (FIG. 1A), a substantially rectangular shape (FIG. 1B, C), etc. It can be a shape. In addition, when the bucket is deep, the adapter cannot be easily attached to and detached from the bucket. Therefore, the adapter may have some play at the corners so that it can be easily taken out, or the handle 3 may be attached to the left or right or the diameter direction of the rotor (FIG. 1C).

  The chamber system receiving portions 2a, 2b, and 2c form recesses and holes that engage with the chamber system 4 in the center portion of the adapter. The chamber system receiving portions 2a, 2b, 2c may be formed as holes (2a, 2b) penetrating in the vertical direction of the adapter 1 when the bottom surface of the bucket of the existing swing rotor is flat and has no irregularities. When the bottom surface of the bucket has irregularities, it is preferable that the chamber system 4 is not stable and is formed as a recess (2c) so that the bottom of the chamber system 4 does not directly contact the bucket.

  In addition, when the swing rotor starts to be centrifuged, a strong centrifugal force is applied outward (in the direction of the arrow in FIG. 2), and the chamber system 4 stands upright during the rotation. Therefore, the direction in which the chamber system is installed is important. In other words, if the installation direction of the chamber system 4 is wrong, a specimen containing cells that have moved downward according to gravity is not applied well on the slide glass even if a centrifugal force is applied, and a clean cell smear cannot be produced. .

  In the case of the auto smear, the projection 5 of the chamber system 4 is formed in the same direction as the lid of the container in which the cell suspension is put. The adapters 1a, 1b, and 1c are also formed with recesses 6a, 6b, and 6c for engaging the protrusion 5, and the adapter 1 is installed so that the protrusion 5 of the chamber system 4 is in the outer peripheral direction of the centrifuge. Good (see FIG. 2). If the installation direction of the adapter 1 is wrong, a clean smear cannot be produced as described above. Therefore, it is preferable to attach a clear marker so that the installation direction of the adapter 1 is not mistaken. For example, in the case of auto smear, characters such as “out” and “out” may be imprinted on the adapter surface so that the projection of the chamber system comes outside the swing rotor in the axial direction.

  FIG. 2 shows a swing rotor R in which four buckets B each having a hexagonal bottom shape are suspended. Although the bucket B in which the adapter 1 is mounted at one place is described, it is needless to say that the bucket B and the adapter 1 need to be installed at all four places, and the weights must be equalized and centrifuged. Here, the adapter 1 is shown in which one chamber system 4 is engaged with one bucket B. However, depending on the size of the bucket, a chamber in which two chamber systems can be installed in one adapter. A system receiver may be formed.

[Example 2] Assembled adapter Next, an assembled adapter according to a second embodiment of the present invention will be described (Fig. 3). The assembly-type adapter 1d includes a basic unit 7 having a chamber system receiving portion 2d and a block-shaped adjustment unit 8 that adjusts the size in an assembly manner according to the size of the centrifuge bucket. As long as the centrifuge bucket is small, the basic unit 7 can be used as an adapter as it is, but when the centrifuge bucket is large, the adjustment unit 8 is fitted and used.

  The adjustment unit 8 has a plurality of fitting projections 9 formed on one surface. On the opposite surface or side surface of the adjustment unit 8 where the fitting projections 9 are formed, fitting recesses 10 are formed at positions corresponding to the fitting projections or at the same interval (FIG. 3B, 3C). A fitting recess 10 is formed at a position corresponding to the fitting projection 9 of the adjustment unit on the bottom and side portions of the basic unit 7, and depending on the size of the bucket, on the side or bottom of the basic unit. By fitting the adjustment unit, the assembly type adapter 1d can be assembled according to the size of the bucket.

[Example 3] Chamber holder directly mounted on a swing rotor Next, a chamber holder 11 which is a third embodiment of the present invention and is directly mounted on a rotor will be described (Fig. 4).

  Depending on the shape of the swing rotor, the chamber holder 11 on which the chamber system 4 is installed may be directly mounted on the rotor R without using a bucket (see FIG. 5). The chamber holder 11 is provided with a chamber system installation unit 12 for installing the chamber system 4. In the case of a chamber holder for auto smear, the chamber system installation part 12 is provided with a recess 13 so as to engage with the protrusion 5 of the chamber system 4. By setting the depth of the chamber system installation part 12 to about 5 mm to 30 mm, the chamber system 4 can be easily attached and detached, and the chamber system does not come off during centrifugation.

  A U-shaped support member 14 is bonded to the bottom of the chamber system installation unit 12. A pair of swing rotor bridging portions 15 are bonded to the upper portion of the support member 14 so that they can be directly mounted on the swing rotor (see FIG. 5).

  The chamber holder 11 may be manufactured to have a width equivalent to that of a bucket of a general-purpose swing rotor and can be mounted as it is. However, the length of the swing rotor bridging portion 15 and the support member 14 are as follows. The width of the chamber holder 11 may be adjusted so that the width of the chamber holder 11 can be adjusted.

  The swing rotor bridging portion 15 may be provided with an elastic body such as a spring at the end so that the length in the lateral direction is adjusted and the swing rotor bridging portion 15 can be stably mounted on the swing rotor (FIG. 4B). A spring portion 16 is provided inside the swing rotor bridging portion 15, and the lateral length can be adjusted by pressing the pressing portion 17. The spring pushes outward, so it can be firmly attached to the swing rotor.

  In addition to the spring, the swing rotor bridging portion 15 is a double cylinder, the outer cylinder is threaded, the inner cylinder is threaded, and the length of the bridging portion is adjusted with screws according to the size of the swing rotor. It is good also as a structure which can be adjusted.

  Furthermore, it is good also as a structure which can adjust a supporting member to the width direction (FIG. 4C). In the case where the length of the support member is adjustable in the lateral direction, the adjustable support members 14a and 14b are a pair of opposed ones having a slit in the center at the bottom of the chamber system installation unit 12. Let it be an L-shaped member. The chamber system installation part 12 is fixed by crimping with an appropriate width by a screw 18 inserted in a slit at the center of the adjustable support members 14a and 14b.

  The adjustment of the lateral width may be a configuration in which the adjustment of the length of the swing rotor bridging portion 15 described above and the adjustment by the support members 14a and 14b may be performed independently, or a configuration in which both are used together. By adopting a configuration in which the lateral width can be adjusted, it can be used in a plurality of types of centrifuges.

  Further, a weight (not shown) can be bonded to a portion near the center of the centrifuge at the lower portion of the chamber setting portion 12 or the lower portion of the support member 14. By applying the weight, the chamber system rotates upright immediately after the start of centrifugation, so that a clean smear can be produced. Even if it is a structure other than the weight, any structure may be used as long as the weight of the portion located inside the swing axis when stationary is heavy with respect to the portion located outside.

[Example 4] Chamber holder directly mounted on a swing rotor Next, another embodiment (FIG. 6) of a chamber holder mounted on a swing rotor according to a fourth embodiment will be described.

  The chamber holder 21 is configured to bend the pair of left and right support members 23a and 23b above the chamber system installation portion 22 and match them at one place at the upper end. A hole 25 through which the swing rotor bridging member 24 for bridging the swing rotor R is formed at a place where the left and right support members 23a and 23b are combined. The swing rotor bridging member 24 passes through the holes 25 of the support members 23a and 23b. By placing the swing rotor bridging member 24 on the swing rotor, the chamber holder 21 can be installed on the swing rotor. The swing rotor bridging member 24 can be installed on swing rotors having different sizes by preparing members having various lengths. Here, the pair of support members 23a and 23b are configured to adhere to the side surface of the chamber system installation unit 22, but may be manufactured as an integral member and bonded to the side surface and the bottom surface of the chamber system installation unit. Further, the support member may be configured as a single belt-like member so as to pass through the center of the chamber system installation portion.

[Example 5] Analysis results using a centrifuge for preparing urine sediments The results of a comparative examination between a smear prepared using the adapter of the present invention and a smear prepared using the auto smear method are shown.

  Samples were prepared using leukocyte suspensions and analyzed to see if the morphology of the cells, particularly the nucleus and cytoplasm, could be identified by Giemsa staining. A leukocyte suspension prepared to a cell number of 100 / μL was prepared using an auto smear sample preparation kit (chamber system, manufactured by Sakura Finetech Japan). A chamber for 1 mL was used, and the chamber, slide glass, and filter paper were set according to a conventional method, and 500 μL of the sample was dispensed. The centrifuge is a biochemical sample measurement centrifuge Kokusan H-80R, and the adapter of the present invention (see Fig. 1B) is installed in the metal bucket holder of the centrifuge, and the chamber system is laid with the lid facing outward. Place gently and centrifuge at 800 rpm for 3 minutes. The rotation speed and time may be the rotation speed and time suitable for the specimen. After completion of the centrifugation, the slide glass was completely dried and fixed with cold air from a dryer.

  Giemsa staining was performed, and staining properties and cell morphology were compared with specimens prepared using an auto smear centrifuge (auto smear method). Microscopic observation is performed at 400 times. The results are shown in FIG. In the figure, black arrows indicate polymorphonuclear cells, white arrows indicate mononuclear cells, and triangles indicate red blood cells. The smear sample (adapter method) prepared using the adapter of the present invention had a clear nucleus and cytoplasm and was able to perform cell differentiation well like the smear sample by the auto smear method.

[Example 6] Results of analysis using a centrifuge for biochemical sample measurement The number of cells and the ratio of disrupted cells were analyzed using a leukocyte suspension. Biochemical specimen measurement centrifuge Kokusan H-80R was used, the chamber was fixed with an adapter to prepare a smear, and compared with the auto smear method. The number of cells in the leukocyte suspension was 100 cells / μL, and 500 μL was dispensed into the chamber to prepare a specimen. A 20-field photograph was taken at 200 times so that the smeared portion of the specimen subjected to Giemsa staining did not overlap on the diagonal line, and four technicians calculated the number of cells and obtained the variation. In addition, the ratio of the cells in which the cell morphology was disintegrated was calculated as disintegrated cells. The results are shown in Table 1.

  There was no significant difference in the number of cells or the ratio of disrupted cells between the adapter method using the adapter of the present invention and the auto smear method.

  As described above, by using the adapter and the chamber holder, it was possible to produce a smear similar to the case using a smearing device such as auto smear or cytospin, using a general-purpose centrifuge. It is a simple method by simply dispensing the sample and centrifuging it, and it does not require special techniques, so even a technician with little experience in smear preparation such as on a day shift can make stable preparations. it can. The cell collection method is used not only for the preparation of smears of cerebrospinal fluid specimens but also for body cavity fluids such as urine, pleural effusions and ascites, and further studies on peritoneal dialysis drainage. The adapter and chamber holder of the present invention can be applied to any centrifuge having a metal bucket, and it is considered that the application of smear preparation with various specimens can be expanded to contribute to clinical practice.

  DESCRIPTION OF SYMBOLS 1a, 1b, 1c ... Adapter, 1d ... Assembling type adapter, 2a, 2b, 2c ... Chamber system receiving part, 3 ... Handle, 4 ... Chamber system, 5 ... Projection, 6a, 6b, 6c ... Recess, 7 ... Basic unit , 8 ... Adjustment unit, 9 ... Fitting convex part, 10 ... Fitting concave part, 11 ... Chamber holder, 12 ... Chamber system installation part, 13 ... Recessed part, 14, 14a, 14b ... Support member, 15 ... Swing rotor bridging part , 16 ... Spring part, 17 ... Pressing part, 18 ... Screw, 21 ... Chamber holder, 22 ... Chamber system installation part, 23a, 23b ... Support member, 24 ... Swing rotor bridging member, R ... Rotor, B ... Bucket

Claims (6)

A centrifuge swing rotor bucket adapter,
An adapter comprising a chamber system receiving portion for engaging a chamber system for producing a smear for cytodiagnosis.   The adapter according to claim 1, wherein a marker for distinguishing between an outer side and an inner side of the swing rotor is provided on the adapter. The adapter includes
The adapter according to claim 1, further comprising a handle for attaching to and detaching from the bucket. The adapter is
A basic unit comprising the chamber system receiver;
The adapter according to claim 1 or 2, comprising an adjustment unit fitted to the basic unit. A chamber holder for installing a chamber system for preparing a smear for cytodiagnosis on a swing centrifuge,
A bridging portion for bridging the swing rotor;
A chamber holder provided with a chamber system installation part. The chamber holder according to claim 5,
A chamber holder configured so that a weight of a portion located inside a swing shaft is heavier than a portion located outside when the swing shaft is stationary.