WO2020171693A1

WO2020171693A1 - Apparatus for immunohistochemical processing of multiple histological samples

Info

Publication number: WO2020171693A1
Application number: PCT/MX2019/000072
Authority: WO
Inventors: Nicole Stephanie MUÑOZ FILIPPETTI; Jesús Raúl BELTRÁN RAMÍREZ; Ma. Del Rocío MACIEL ARELLANO; Javier ESPINOZA JR.; Víctor Manuel LARIOS ROSILLO; José Ángel DOMÍNGUEZ RAMÍREZ; Jonathan De Jesús MARTÍNEZ MENDOZA; Elsa Julieta CEDILLO ELÍAS; Guillermo GARCÍA TORALES
Original assignee: Consorcio De Investigación E Innovación Abierta S.A.P.I. De C.V.
Priority date: 2019-02-21
Filing date: 2019-06-06
Publication date: 2020-08-27
Also published as: MX2019002085A

Abstract

The present invention refers to an apparatus for immunohistochemical processing of multiple histological samples, which can process biological (histological) samples; said apparatus has a main structure with a platform at the top which has an opening at the centre and channels over which the reagent to be used is distributed from a container inside the structure, and by means of a pump, the reagent is capable of reaching trays, which are attached to the main structure by means of bolts that allow them to move in different directions, with the intention of applying movement to the process and so that the samples are covered by the reagents. The main advantage of the apparatus for immunohistochemical processing of multiple histological samples is that more than one sample can be processed at the same time, thus reducing processing times in serial samples.

Description

APPARATUS FOR THE IMMUNOHISTOCHEMICAL PROCESSING OF MULTIPLE HISTOLOGICAL SAMPLES

TECHNICAL FIELD OF THE INVENTION

The present invention is related to the technical field of medicine, mechanics, electronics since it provides an apparatus for the immunohistochemical processing of multiple histological samples.

BACKGROUND OF THE INVENTION

The first histological investigations were possible from the year 1600, when the microscope was incorporated into anatomical studies. Marcello Malpighi is the founder of histology and his name is still linked to various histological structures. In 1665 the existence of small units within the tissues was discovered and they were called cells. In 1830, accompanying the improvements that were introduced in optical microscopy, it was possible to distinguish the cell nucleus. In 1838 the concept of cell theory was introduced.

In the following years, Rudolf Virchow introduced the concept that every cell originates from another cell (omnis cellula ex cellula).

The modern technological development of research tools allowed an enormous advance in histological knowledge. Among them we can mention electron microscopy, immunohistochemistry, and in situ hybridization technique. Recent techniques added to new research gave way to the emergence of cell biology.

Immunohistochemistry is a procedure that aims to detect, amplify, and make a specific antigen visible, which is generally a protein. This technique makes it possible to identify the location of a specific substance at the tissue or cellular level. It is based on the use of antibodies that specifically bind to a substance to be identified (primary antigen). These antibodies may have an enzyme attached or it may be attached to a secondary antibody that recognizes and binds to the primary one.

Applied to an organic tissue, the primary antibody specifically binds to the substrate and the enzymatic activity is exploited to visualize the binding. In this way, a substrate-antibody-enzyme complex is achieved bound to the place where the substrate is located and by activating the enzyme with the addition of its substrate, an identifiable product is generated where the complex is found.

This technique makes it possible to identify the exact location of a tissue or cytological substance, identifying the antigenic markers characteristic of a cell line, cells that secrete a protein, membrane receptors, tissue concentration gradients or cells that have responded to a hormone (with specific antibodies for intracellular signaling pathways).

This procedure is also used in histology, which is the discipline that studies everything related to organic tissues: their microscopic structure, their development and their functions. Histology is identified with microscopic anatomy, since its study does not stop at the tissues, but goes further, also observing internally cells and other corpuscles, relating to biochemistry and cytology. Histology has several subdivisions that allow to improve the study approach, thus, there is general histology that is responsible for the study of basic tissues and histology of systems that is responsible for the study of the tissue structure of devices and systems; In addition to other approaches by regions such as oral histology that is responsible for carrying out the conjunction of the general histology approach and the systems to talk about a specific region and understand the tissue composition and development, being able to make a greater focus towards the dental organ 1

Histology has never had the prominence in the biology and medicine curriculum that it has achieved today. Histology is the study of the microscopic structure of biological material and the way in which the various individual components are related both structurally and functionally. It is crucial for medicine and for biology because it lies at the intersections between biochemistry, molecular biology and physiology on the one hand and pathological processes and their consequences on the other.

Histologists pay increasing attention to chemical problems. Thus, for example, the aspiration to determine exactly the chemical composition of certain structures of the living mass is spreading among them, when studying enzymes, ions, proteins, carbohydrates, fats and lipoids, ferments, etc. in cells and tissues with the aid of the microscope.

A search of the state of the art was carried out to provide devices for the immunohistochemical processing of histological samples, where it was found that different equipment has been developed, as mentioned in the patent application document number US2009004691 (Al) (Al), with Publication date of January 1, 2009, entitled "IMPROVED FLUID METHOD AND APPARATUS FOR RAPID AUTOMATED IMMUNOHISTOCHEMISTRY", which describes a sample processing system that can be configured to achieve rapid sample processing, such as histochemical processing fast, it may involve a wave element that can use angular microscopic sliding motions to cause repeated removal and reapplication of a fluid substance perhaps through the action of capillary motion to update a microenvironment adjacent to a sample, such as a biopsy or another similar sample. With such cooling of a microenvironment, depletion of the microenvironment is avoided and the time required for slide processing can be drastically shortened from the more common 60 to 120 minutes to perhaps less than 15 minutes to allow use of such a system in an intraoperative device or surgical setting as recommended by the intraoperative guidelines of the College of American Pathologists or similar; therefore, patients can avoid the need for an additional surgical procedure when laboratory results are available to see if tumors or the like were completely removed in a previous procedure. The above-described document aims to decrease reaction, binding or interaction times through a system that contains particular conditions within itself, allowing a microenvironment to be supplied in an external sample area so that in this way a next reaction can be carried out. out faster. However, there is no evidence that it can process more than one sample at the same time and thus reduce the times of serial samples, the invention described does not have a set of trays that have the ability to place more than one sample at the same time. , does not describe a reagent container and a hydraulic pump that allows the transfer of the same to the samples through the use of channels located from the center of an upper base to the trays, Nor does it present a control module and a graphical interface that allows the set the activation times for both the reagent pump and the movement of the motors. Another document that was found is patent number US9040284 (B2), with publication date of May 26, 2015, entitled "SYSTEM AND METHOD OF PROCESSING SAMPLES AND TEMPERATURE CONTROL", which describes systems and methods of sample processing and temperature control. The invention disclosed in the document may be especially related to temperature control, and in some embodiments may be methods of controlling the temperature of an automated sample processing system and automated sample processing methods. Specifically, it provides temperature control in relation to sample processing systems and sample processing methods, and in some embodiments provides temperature control in relation to with sample carriers and processing materials such as reagents. Corresponding systems and devices are described, including sample processing systems, sample carrier temperature regulation systems, reagent temperature regulation systems, sample processing control systems, and devices. temperature regulation, among other embodiments. As can be seen in document US9040284 (B2), the main characteristic of this invention is that it is equipped with a temperature control in relation to the automatic processing of samples and the objective of this is to analyze histological samples. However, its main structure does not have a hole in the upper part through which the reagent to be used is distributed from a container within the structure itself, nor does it describe having trays that can be moved in different directions and to which reagent is targeted, thereby allowing samples to be covered by reagents.

Finally, the publication of application number EP1685382 (A2) was found, with publication date of August 2, 2006, which is entitled "Universal multiaxial testing machine", said application refers to a universal multiaxial testing machine , which allows evaluating the mechanical behavior and performance of materials with flat structures, such as fabrics, composites and laminates. The machine comprises 4 horizontal axes at 45 °, with 8 movable gripping jaws along the sliding rails and driven by the action of 8 independent motors. The connection between a gripping jaw and its respective motor is ensured by a linear actuator. The test specimen is clamped with the gripping jaws and can be subjected to tensile, compression and fatigue tests, making it possible to analyze the behavior of materials under simultaneous multidirectional loads.

As can be seen, the document describes an invention whose objective is to carry out physical tests on materials, therefore, the mechanism it presents forms a set of fasteners and motors connected to these, with the help of a linear guide, the material to be subjected to the tests, it remains in one position, and there is no evidence that it can process biological (histological) samples, lacking a functional structure for this purpose, such as a main structure with a hole in its upper part where the reagent to be used from a container inside the structure, which with the help of a pump, the reagent is able to reach some trays, which are attached to the main structure by means of a joint that allows them to move in different directions, with the intention of applying movement to the process and the samples are covered by the reagents.

In the documents previously described, it is not evidenced that the equipment provides a main structure that has an orifice in the upper part through which the reagent to be used is distributed from a container within the structure itself, nor does it describe that it has multiple trays that can be moved in different directions and to which the reagent is directed, thus allowing the samples to be covered by the reagents, which allows to process more than one sample at the same time and thus reduce the times of serial samples.

OBJECT OF THE INVENTION

It is, therefore, object of the present invention, to provide an apparatus for the immunohistochemical processing of multiple histological samples, which solves the aforementioned problems.

BRIEF DESCRIPTION OF THE FIGURES

The characteristic details of this novel apparatus for the immunohistochemical processing of multiple histological samples are clearly shown in the following description and in the accompanying figures, as well as an illustration thereof, and following the same reference signs to indicate the parts shown. However, these figures are shown by way of example and should not be considered as limiting for the present invention.

Figure 1 shows a perspective view of the apparatus for immunohistochemical processing of multiple histological samples.

Figure 2 shows a perspective view of the apparatus for immunohistochemical processing of multiple histological samples, with the transparency of the main structure.

Figure 3 shows a perspective view of the apparatus for immunohistochemical processing of multiple histological samples, without the main structure. Figure 4 shows a perspective view of the main structure of the apparatus for immunohistochemical processing of multiple histological samples.

Figure 5 shows a perspective view of the platform of the apparatus for immunohistochemical processing of multiple histological samples.

Figure 6 shows a detailed perspective view of the tray of the apparatus for immunohistochemical processing of multiple histological samples.

Figure 7 shows a detailed perspective view of the spherical support installed in the tray of the apparatus for the immunohistochemical processing of multiple histological samples.

Figure 8 shows a detailed perspective view of the bolt of the apparatus for immunohistochemical processing of multiple histological samples.

Figure 9 shows a perspective view of the internal structure of the apparatus for immunohistochemical processing of multiple histological samples.

Figure 10 shows a view of the assembly of the support rods with the motor of the apparatus for the immunohistochemical processing of multiple histological samples.

Figure 11 shows a view of the connection of the pipes of the hydraulic pump with the container of the apparatus for processing immunohistochemical analysis of multiple histological samples.

Figure 12 shows a top view of the apparatus for immunohistochemical processing of multiple histological samples.

DETAILED DESCRIPTION OF THE INVENTION

For a better understanding of the invention, the parts that make up the apparatus for the immunohistochemical processing of multiple histological samples are listed below:

1. Main structure

2. Slot

3. Platform

4. Orifice

5. Channel

6. Internal structure

7. Post

8. Support

9. Hydraulic pump

10. Pipe

11. Container

12. Bolt

13. Tray

14. Relief

15. Entrance

16. Elevation

17. Spherical support

18. Support rods

19. Engine

20. Control module

21. Display With reference to the figures, the apparatus for the immunohistochemical processing of multiple histological samples is made up of a main structure (1) in a prismatic shape, preferably octagonal, which has at least one groove in the upper part of its walls ( two); In the upper part of the main structure (1) a platform (3) is placed which is in the same prismatic shape as said main structure, the edges of the platform are aligned with the edges of the main structure (1); The platform (3) has a hole (4) in the center and, at least, one channel (5) for each wall of the main structure (1), these channels function as reagent guides to carry out the immunohistochemical processing of the histological samples.

An internal structure (6) that has a cylindrical shape, is configured to be placed inside the main structure (1) in its upper part, as can be seen in figure 2. The internal structure (6) has a post (7) For each wall of the main structure (1) which is positioned adjacent to the grooves (2) of said main structure (1), a support (8) is located in the lower part of the internal structure (6), on the which is placed a hydraulic pump (9) which has a pipe

(10) connected to the hole (4) of the platform (3) on one side and on the other side it is connected to at least one container

(11) filled with reagents used for histological tests; This configuration allows that when the hydraulic pump (9) is activated the reagent leaves through the orifice (4) and flows through the channels (5) of the platform (3). At the center of each edge of the platform (3) is placed at least one bolt (12) which is configured to rotate about the horizontal by means of its axis. As illustrated in figure 7, the bolts (12) have a channel on their surface which coincides with the channel (5).

On each bolt (12) a tray (13) is placed as the one illustrated in figure 6, said tray is prismatic in shape, preferably rectangular and has an unevenness (16) which has reliefs (14) in which you can place the samples to be analyzed. The end of the trays (13) that is attached to the bolt (12) has an inlet (15) in the form of a channel that coincides with the channel of the bolt (12) and the channels (5) of the platform (3); This configuration allows the reagent that comes out of the hole (4) and flows through the channels (5) of the platform (3), passing through the channel of the bolt (12) and the inlet (15) of the tray ( 13) to fill the gap (16) covering all the samples, to carry out the immunohistochemical processing.

The trays (13) have on their underside a spherical support (17) configured to rotate and move vertically; On said spherical supports (17) the end of a support rod (18) is placed, and the opposite end of the support rod (18) passes through the slot (2) of the main structure (1) and is fastened to a motor (19) that is fixed on each post (7) of the internal structure (6); This configuration allows the tray (13) to be tilted and perform oscillatory movements, making the process of filling the unevenness (16) with the reagent more efficient and uniform. Once the reagent was in contact with the samples for the established time, the trays (13) are raised vertically, causing the reagent to return through the inlet (15) and the channels (5) of the platform (3) until it reaches the hole (4 ), in such a way that the reagent returns to the container (11) to be used again in other tests.

On the outside of the main structure (1), a control module (20) is placed consisting of a microcontroller (not illustrated) that is connected to the motors (19) and the hydraulic pump (9), to drive them accordingly. with previously programmed parameters. The control module (20) has a touch screen (21), which preferably is liquid crystal, and shows the information of the immunohistochemical process of histological samples that is being carried out, by means of the screen (21) you can program the microcontroller with the parameters that the apparatus works at for the immunohistochemical processing of multiple histological samples.

PREFERRED EMBODIMENT OF THE INVENTION

Examples

The following examples illustrate a preferred manner of how to carry out the embodiment of the present invention, so they should not be considered as limiting it.

Example 1. Operation of the apparatus for immunohistochemical processing of multiple histological samples.

The histological samples are placed in the reliefs (14) of the trays (13), subsequently by means of the screen (21) the working parameters are programmed, allowing the microcontroller (not illustrated) to actuate the hydraulic pump (9) so that the reagent found in the container (11), flows through the pipe (10) and out through the orifice (4) and by means of the pressure travel through the channels (5) of the platform (3) passing through the entrance (15) of the trays (13) while the motors (19) are also driven by the microcontroller to transmit the vertical and oscillatory movement of the support rods (18), allowing the trays (13) to move until the reagent fills the gap (16) and covers all of the samples.

Once the reagent was in contact with the samples for the established time, the trays (13) are raised vertically, causing the reagent to return through the inlet (15) and the channels (5) of the platform (3) until reach the hole (4), in such a way that the reagent returns to the container (11) to be used again in other tests.

The invention has been described sufficiently so that a person of average knowledge in the matter can reproduce and obtain the results mentioned in the present invention. However, any person skilled in the field of the art that concerns the present invention may be able to make modifications not described in the present application, however, if for the application of these modifications in a certain structure or in the manufacturing process of the same, the matter claimed in the following claims is required, said structures should be understood within the scope of the invention.

Claims

1. An apparatus for the immunohistochemical processing of multiple histological samples, characterized in that it comprises: a main structure (1) in a prismatic shape, which has at least one slot in the upper part of its walls

(two); In the upper part of the main structure (1) a platform (3) is placed which is of the same prismatic shape as said main structure (1), the edges of the platform are aligned with the edges of the main structure (1 ); the platform

(3) has a hole

(4) in the center and at least one channel

(5) for each wall of the main structure (1), said channels function as guides of the reagent to carry out the immunohistochemical processing of the histological samples; an internal structure (6) is configured to be placed inside the main structure (1) in its upper part; the internal structure

(6) has a pole

(7) for each wall of the main structure (1), said post is positioned adjacent to the grooves (2) of said main structure (1); a support

(8) is located in the lower part of the internal structure (6), on which a hydraulic pump is placed

(9) that has a pipe (10) connected to the hole (4) of the platform (3) on one side and on the other side it is connected to at least one container (11) filled with reagents used for histological tests ; at the center of each edge of the platform (3) is placed at least one bolt (12) that is configured to rotate on the horizontal through its axis, said bolts (12) have on their surface a channel which coincides with the channel (5); On each bolt (12) a tray (13) is placed that has a slope (16) which has reliefs (14) in which they can place the samples to be analyzed; the end of the trays (13) that is attached to the bolt (12) has an inlet (15) in the form of a channel that coincides with the channel of the bolt (12) and the channels (5) of the platform (3); the trays (13) have on their underside a spherical support (17) configured to rotate and move vertically; On said spherical supports (17) the end of a support rod (18) is placed, and the opposite end of the support rod (18) passes through the slot (2) of the main structure (1) and is fastened to a motor (19) that is fixed on each post (7) of the internal structure (6), this configuration allows the tray (13) to tilt and perform oscillatory movements; On the outside of the main structure (1), a control module (20) made up of a microcontroller (not illustrated) is placed that is connected to the motors (19) and the hydraulic pump (9), to drive them according to with previously programmed parameters; and, the control module (20) has a screen (21) by means of which the microcontroller can be programmed.

2. The apparatus of claim 1 characterized in that the prismatic shape of the main structure (1) is preferably octagonal.

The apparatus of claim 1, characterized in that the internal structure (6) is cylindrical in shape.

4. The apparatus of claim 1 characterized in that the tray (13) is prismatic in shape.

5. The apparatus of claim 4 characterized in that the prismatic shape is preferably rectangular.

6. The apparatus of claim 1 characterized in that the screen (21) is tactile. 7. The apparatus of claims 1 and 6 characterized in that the screen (21) is preferably made of liquid crystal.