Portable device for multiplex staining and analysis of histopathology samples

Abstract

The present invention relates to a portable device for the staining and analysis of histopathology samples, formed by a main base on which cylindrical structures are arranged, said structures containing containers of slides. The slides are individually stained, and the reagent can be reutilised by means of a two-way pump that returns the reagent to the container once used. In order to stain only one sample per container, the device comprises a circular base provided with a rectangular opening that allows the reagent to pass from a reagent container to the slide container; if another sample needs to be stained, a stepper motor is used to rotate the cylindrical structure such that another slide container is positioned above the rectangular opening. A camera obtains images for the analysis of the stained cytology or histology samples. A smart device or a screen located on a side of the main base can be used to configure the operation of the device of the invention and view the results obtained.

Description

[0001] PORTABLE DEVICE FOR MULTIPLE STAINING AND ANALYSIS OF HISTOPATHOLOGICAL SAMPLES

[0002] TECHNICAL FIELD OF THE INVENTION

[0003] The present invention relates to the technical fields of mechanics, electronics, biomedical engineering, microfluidics, biotechnology, diagnostic medicine, and more specifically to cytology, as it provides a portable device for multiple staining and analysis of histopathological samples. This device can stain and analyze several slides with cytological and histological samples within cylindrical containers on a main base, using one camera per cylinder to analyze several samples simultaneously. Various containers and hoses are used to deliver the reagent to each container, utilizing bidirectional pumps and valves that also allow the reagent to be returned to its container for reuse.

[0004] BACKGROUND OF THE INVENTION

[0005] Histopathological samples, which include both cytological and histological samples, are fundamental for the diagnosis and study of a wide range of diseases at the cellular and tissue levels. These samples allow direct observation of the morphological changes that occur in cells and tissues, which is essential for identifying pathologies such as cancer, infections, and inflammatory diseases.

[0006] Cytological samples focus on the analysis of individual cells obtained through various methods, such as smears, aspirates, or washings. These samples are crucial for detecting abnormalities in cell morphology that may indicate the presence of disease. For example, a Pap smear is used for the early detection of cervical cancer, while cytology of bodily fluids, such as pleural or peritoneal fluid, can reveal the presence of malignant or infectious cells.

[0007] Histological samples, on the other hand, involve the study of thin sections of whole tissue. These samples provide a more comprehensive view of tissue architecture and the relationships between different cell types and the extracellular matrix. Histological staining techniques, such as hematoxylin and eosin (H&E) staining, are widely used to differentiate cell nuclei from cytoplasm and to highlight different tissue types. Other techniques, such as immunohistochemistry, allow the identification of specific proteins in cells, providing detailed information on molecular expression and helping to better characterize diseases at the histological level.

[0008] The histopathological analysis of these samples involves evaluating various morphological aspects to identify pathological characteristics. In cytological samples, this may include observing atypical cells, changes in cell size and shape, and the presence of intracellular inclusions. In histological samples, the analysis may focus on the overall tissue structure, cellular organization, the presence of inflammation, necrosis, fibrosis, and abnormal cell proliferation.

[0009] The accuracy of histopathological diagnosis depends largely on the quality of the samples, the preparation and staining techniques used, and the pathologist's skill in interpreting the findings. Both cytological and histological samples play complementary roles in medical diagnosis. While cytological samples allow for rapid and less invasive analysis, histological samples provide a more detailed and comprehensive understanding of tissue architecture.

[0010] Staining is an auxiliary technique used in microscopy to enhance contrast in the image viewed under the microscope. Dyes and dyes are substances commonly used in biology and medicine to highlight structures in biological tissues that will be observed with different types of microscopes. Different dyes can be used to increase definition and examine large tissue sections (highlighting, for example, muscle fibers or connective tissue), cell populations (for example, classifying different blood cells), or even to highlight organelles within individual cells.

[0011] Different types of biological stains are also used to label cells in flow cytometry and to label proteins or nucleic acids in gel electrophoresis. In its simplest form, the actual staining process may involve immersing the sample (before or after fixation and mounting) in the staining solution, followed by rinsing, which is a washing to remove excess stain, and observation.

[0012] When working in the field of cytology and histology, the need to stain various cytological and histological samples is very common. This requires different types of reagents depending on what needs to be observed in the sample. Furthermore, reagents are often wasted because there is no way to collect them again if they can be reused. Additionally, there are times when samples need to be transported safely from one place to another, or staining equipment needs to be taken to remote locations far from the laboratories, which complicates the work of those in this area. Portable devices for multiple staining and analysis of histopathological samples have revolutionized the field of clinical diagnosis and biomedical research. Traditionally, the staining and analysis of histopathological samples was carried out in fixed laboratories, requiring bulky and specialized equipment.This approach not only limited accessibility to these services in remote regions or in emergency situations, but also increased the waiting time for crucial diagnostic results.

[0013] The development of portable multistaining devices arose in response to the need for rapid and efficient point-of-care cytological analysis. These devices are designed to be compact, lightweight, and easy to use, enabling healthcare professionals to perform diagnostic procedures directly at the patient's location. This point-of-care testing capability is especially useful in rural areas, conflict zones, or during epidemic outbreaks, where access to well-equipped laboratories may be limited or nonexistent.

[0014] Technological innovation in these devices has enabled the integration of multiple stages of the staining and analysis process into a single portable unit. Advanced microfluidic techniques and the development of standardized staining reagents have facilitated the miniaturization of these systems without compromising the accuracy and reproducibility of the results. Furthermore, the integration of automated detection systems and AI-based analysis algorithms has significantly improved the ability of these devices to provide rapid and reliable diagnoses.

[0015] Historically, staining histopathological samples has been a crucial technique in the diagnosis of various diseases, including infections, inflammatory diseases, and cancer. Multiplex staining, which uses several dyes to highlight different cellular components, offers a detailed view of cell morphology and pathological characteristics. Portable devices have made it possible to adapt these complex staining techniques into accessible and manageable formats, while maintaining the quality of diagnoses traditionally obtained in specialized laboratories.

[0016] The impact of portable multistaining and histopathological sample analysis devices extends beyond clinical diagnosis. These devices have also found applications in biomedical research, where their portability and efficiency allow researchers to conduct field studies and obtain data in real time. This is particularly relevant in epidemiological studies and clinical trials, where the speed and accuracy of sample collection and analysis can significantly influence study results and clinical decision-making.

[0017] Portable multistaining and histopathological analysis devices represent a significant advance in diagnostic medicine and biomedical research. By providing an accessible and efficient solution for point-of-care diagnostic testing, these devices have improved the healthcare system's responsiveness to diverse clinical situations and democratized access to high-quality diagnostics worldwide.

[0018] Various devices and systems have been developed to perform staining on cytology samples, such as the apparatus described in US patent 11,926,815 (B2), which outlines a system, apparatus, and method for collecting and preparing individual cells, nuclei, subcellular components, and biomolecules from samples, including tissues, and in some embodiments, using the individual cells to form organoids or microtissues. The system can perform enzymatic and / or physical disruption of the tissue to dissociate it into individual cells and then use a hanging drop method to form organoids or microtissues.

[0019] Also included is the system and method disclosed in patent CA2745561(C), which describes an apparatus comprising at least one staining module and one coating module; an imaging module; a storage module; an automated transport module for transporting at least one film between at least one of the staining and coating modules, the imaging module, and the storage module; and a controller. A method comprising processing at least one film; determining whether an imaging module is available for imaging a biological sample on at least one film; transporting at least one film to the imaging module using an automated transport module; and transporting at least one film to a storage module using the automated transport module when the imaging module is determined to be unavailable.A system that includes a processing module for processing at least one sheet with a biological sample.

[0020] As will be observed later in the detailed description of the invention, there are differences between the apparatus and systems developed in the patents mentioned, differences that provide technical advantages for the analysis of multiple cytological and histological samples by staining, in a single device.

[0021] OBJECT OF THE INVENTION

[0022] The present invention aims to provide a portable device for multiple staining and analysis of histopathological samples, which can stain and analyze several slides with cytological and / or histological samples within cylindrical containers on a main base, using one camera per cylinder to analyze several samples at the same time. In addition, there are various containers and hoses to carry the reagent to each container using bidirectional pumps and valves that also allow the reagent to be returned to its container for reuse. Additionally, there is a screen to visualize the analysis, pins for data transmission, and charging pins to power the camera and its movement mechanism.

[0023] BRIEF DESCRIPTION OF THE FIGURES

[0024] The characteristic details of this novel portable device for multiple staining and analysis of histopathological samples are clearly shown in the following description and accompanying figures, as well as an illustration thereof, using the same reference symbols to indicate the parts shown. However, these figures are shown by way of example and should not be considered limiting to the present invention.

[0025] Figure 1 shows a front perspective view of the portable device for multiple staining and analysis of histopathological samples.

[0026] Figure 2 shows a perspective view of the transparency of the portable device for multiple staining and analysis of histopathological samples, allowing internal details of the main base and hollow vertical protrusion to be seen.

[0027] Figure 3 shows a perspective view of the portable device for multiple staining and analysis of histopathological samples, showing details of the circular cavities.

[0028] Figure 4 shows a perspective view of the internal connections of the tubing and hoses that distribute the reagents of the portable multi-staining device for histopathological sample analysis.

[0029] Figure 5 shows a perspective view of the internal connections of the tubing and hoses that distribute the reagents of the portable multi-staining device for histopathological sample analysis.

[0030] Figure 6 shows a detailed view of the internal connections of the tubing and hoses that distribute the reagents of the portable multi-staining and histopathological sample analysis device. Figure 7 shows a perspective view highlighting details of the cylindrical structure of the portable multi-staining and histopathological sample analysis device.

[0031] Figure 8 shows a perspective view of the reading mechanism of the portable device for multiple staining and analysis of histopathological samples.

[0032] DETAILED DESCRIPTION OF THE INVENTION

[0033] For a better understanding of the present invention, the parts that make up the portable device for multiple staining and analysis of histopathological samples that is the subject of the present invention are listed below:

[0034] 1. Main base

[0035] 2. Cylindrical structure

[0036] 3. Circular cavity 4. Rectangular cut

[0037] 5. Charging pins

[0038] 6. Data transfer pins 7. Lower base

[0039] 8. Camera

[0040] 9. Screen

[0041] 10. Circular perforation

[0042] 11. Hollow vertical projection

[0043] 12. Reagent containers

[0044] 13. Bidirectional pump

[0045] 14. Horizontal tube

[0046] 15. Connector

[0047] 16. Bidirectional valve

[0048] 17. Vertical tube

[0049] 18. Bluetooth Module

[0050] 19. Hose

[0051] 20. Control Module

[0052] 21. Rechargeable battery

[0053] 22. Loading Port

[0054] 23. Flake container

[0055] 24. Magnet

[0056] 25. Circular base

[0057] 26. Rectangular projection

[0058] 27. Stepper motor

[0059] 28. Second magnet

[0060] 29. Rail

[0061] 30. Hinge-type support

[0062] 31. Rectangular perforation

[0063] 32. Protuberance

[0064] 33. Tube

[0065] 34. Flat nozzle

[0066] 35. Support With reference to the figures, the portable device for multiple staining and analysis of histopathological samples that is the subject of the present invention comprises two essential parts, a main base (1) and cylindrical structures (2). The main base (1) is in the form of a hollow rectangular prism. On its upper face are located a plurality of circular cavities (3), each of which serves to hold a cylindrical structure (2). Inside each circular cavity (3), near its perimeter, there is a rectangular cut (4), as can be seen in Figure 3, which allows the passage of the reagent to a container of slides (23) located inside the cylindrical structure (2).Inside each of the circular cavities (3) there is a pair of charging pins (5) which allow the operation of a slide reading mechanism and another pair of information transfer pins (6) located inside each of the circular cavities (3) which are for transferring the information from the slide reading obtained with a camera (8) as illustrated in figure 8. This information is sent to a screen (9) located on one side of the main base (1) attached to it by means of a hinge-type support (30) that allows the rotation of the screen (9).In the center of each of the circular cavities (3) there is a circular perforation (10) that serves to place the cylindrical structure (2), by means of a protrusion (32) located on the lower surface of a base (25) that each cylindrical structure (2) has; this protrusion (32) has the same shape and dimension as the circular perforation (10) allowing the cylindrical structure (2) to be held firmly. The circular base (25) is the size of the cylindrical structure (2); on the upper face of this circular base (25) near its perimeter there is a rectangular perforation (31) that coincides in direction and dimensions with the rectangular cut (4) that is located in the circular cavity (3) of the main base (1).At the rear of the main base (1) there is a hollow vertical projection (11), in which a plurality of reagent containers (12) are placed, each of which at its lower part is connected by a tube (33) to a bidirectional pump (13) located at the bottom of the hollow vertical projection (11). Each of the bidirectional pumps (13) is connected to a horizontal tube (14) that enters the main base (1) and reaches its end. These horizontal tubes (14) serve to distribute the reagent found in each reagent container (12) to each of the cylindrical structures (2) through hoses (19) that are connected to bidirectional valves (16) located inside the main base (1), preferably below each circular cavity (3).

[0067] At the bottom of each circular cavity (3) there is a multi-inlet, single-outlet connector (15), installed diagonally. Each inlet has a bidirectional valve (16), and the outlet is connected to a vertical tube (17) for connection to the cylindrical structure (2). The vertical tube (17) has a rectangular flat nozzle (34) at its end, the size of the rectangular cut (4) in the circular cavity (3), which serves to distribute the reagent to each of the slide containers (23). A hose (19) is connected from each bidirectional valve (16) to each of the horizontal tubes (14), each of which contains the reagent from each of the reagent containers (12).

[0068] The main base (1) houses inside, below the circular cavities (3), a control module (20) and a Bluetooth module (18) connected to it, both powered by a rechargeable battery (21). These elements are mounted on a lower base (7) that covers the entire bottom of the main base (1). On one side of the main base (1) there is a charging port (22) for charging the rechargeable battery (21).

[0069] With reference to figures 7 and 8, each of the cylindrical structures (2) which are hollow, has around the inner face of the same, a plurality of slide containers (23) in the shape of a hollow rectangle to place inside a slide with a cytological or histological sample that is to be stained, these containers are placed vertically and have a perforation on their lower face that allows the passage of the reagent as well as the introduction of the slides with a cytological or histological sample to be stained.

[0070] In the center of the inner upper face of the cylindrical structure (2) there is a magnet (24) preferably circular in shape which allows said cylindrical structure (2) to be joined with a movement mechanism; which is composed of a rectangular projection (26) placed in the center of said circular base (25), said rectangular projection (26) has on its upper part a stepper motor (27) on whose axis there is a second circular magnet (28) which joins with the magnet (24) of the cylindrical structure (2), allowing it to rotate with each movement of the stepper motor (27) and in turn the cylindrical structure (2) also rotates.

[0071] On one of the faces of the rectangular projection (26) is the reading mechanism consisting of a worm screw with a rail (29) held by a support (35) at its upper and lower part on the rectangular projection (26); on one of the supports is a motor (not illustrated) placed inside the rectangular projection (26) for the movement of the worm screw, on the rail (29) is installed a base with a camera ( 8 ) which allows obtaining images for the analysis of the stained cytological or histological samples.This analysis requires scanning the sample. The camera (8) is placed at the base of the rail (29) on top of the screw conveyor, and the cylindrical structure (2) is moved with the help of the stepper motor (27) until the camera (8) scans the width of the stained slide container (23). The camera (8) is then lowered by moving the screw conveyor (29), and the stepper motor (27) rotates the cylindrical structure (2) again, this time in the opposite direction, until it reaches the end of the container. The images captured by the camera (8) are sent to the screen (9), and this information can also be sent to a smart device via the Bluetooth module (18).

[0072] The configuration of the portable device for multiple staining and analysis of histopathological samples, which is the subject of the present invention, allows for the staining of several cytological and histological samples at the same time, as well as their analysis, and allows the reagent to be reused with the implementation of the bidirectional pump (13) that returns the reagent to its corresponding reagent container (12) once it has been used to stain the sample.

[0073] A feature of the portable device for multiple staining and analysis of histopathological samples that is the subject of the present invention is that the rectangular perforation (4) located in the circular base (25) is configured to stain only one sample per slide container (23), since it allows the passage of the reagent coming from the reagent container (12), to then pass through the horizontal tube (14), the hose (19) and the bidirectional valve (16) of the connector (15) with multiple inlets and one outlet, then passes through the vertical tube (17) and through the rectangular perforation (4) until it reaches the slide in a slide container (23).If another sample needs to be stained, the stepper motor (27) is used, which rotates the cylindrical structure (2), positioning another slide container (23) above the rectangular perforation (4). The process is then repeated, sending the reagent from the desired reagent container (12) to the slide container (23). The operation of the portable device for multiple staining and analysis of histopathological samples, which is the subject of this invention, is configured using a smart device via the Bluetooth module (18) or through the screen (9) located on the side of the main base (1).

[0074] In view of the foregoing, it is reiterated that the scope of the present invention shall not be limited by the specific embodiments described, and it shall be understood that variations may be made in several respects. Such variations shall not be considered a departure from the spirit and scope of the invention, and all such modifications may be obvious to a person skilled in the art and shall be included within the scope of the following claims.

Claims

CLAIMS 1. A portable device for multiple staining and analysis of histopathological samples, characterized in that it comprises: a main base (1) in the form of a hollow rectangular prism, which has on its upper face a plurality of circular cavities (3), which in its interior near its perimeter has a rectangular cut (4) which allows the passage of the reagent; a pair of loading pins (5) located within each of the circular cavities (3) which allow the operation of a slide reading mechanism and another pair of information transfer pins (6) located within each of the circular cavities (3) to transfer the information of the reading obtained from the slide;In the center of each of the circular cavities (3) there is a circular perforation (10) that serves to place a hollow cylindrical structure (2) in each perforation, by means of a protrusion (32) that is located on the lower surface of a base (25) that each cylindrical structure (2) has; a hollow vertical projection (11) located at the back of the main base (1), in which a plurality of reagent containers (12) are placed, each of which at its lower part is connected by a tube (33) to a bidirectional pump (13) located at the bottom of the hollow vertical projection (11); each of the bidirectional pumps (13) is connected by a horizontal tube (14) that enters the main base (1) and reaches its end;A multi-input, single-output connector (15) is located at the bottom of each of the circular cavities (3), with a bidirectional valve (16) at each of the inlets and a vertical tube (17) connected to the outlet for connection to the cylindrical structure (2); a control module (20), a Bluetooth module (18) powered by a rechargeable battery (21), are housed inside the main base (1), all mounted on a lower base (7) that covers the entire bottom of the base; main (1); each of the cylindrical structures (2) has around its inner face a plurality of slide holders (23) shaped like a hollow rectangle to place inside a slide with a cytological or histological sample to be stained; in the center of the inner upper face of the cylindrical structure (2) there is a magnet (24) that allows said cylindrical structure (2) to be joined with a movement mechanism, which consists of a rectangular projection (26) placed in the center of the circular base (25), said rectangular projection (26) has on its upper part a stepper motor (27) on whose axis there is a second magnet (28) that is joined with the magnet (24) of the cylindrical structure (2), allowing it to rotate with each movement of the stepper motor (27) and in turn;A rectangular perforation (31) is placed on the upper face of the circular base (25) near its perimeter; this rectangular perforation (31) coincides in direction and dimensions with the rectangular cut (4) located in the circular cavity (3) of the main base (1); a reading mechanism is placed on one of the faces of the rectangular projection (26) and consists of a worm gear with a rail (29) held by a support (35) at its upper and lower parts on the rectangular projection (26); a motor is located inside the rectangular projection (26) on one of the supports for the movement of the worm gear; a base with a camera (8) is installed on the rail of the worm gear (29), which allows obtaining images for the analysis of stained cytological or histological samples; and a screen (9) is located on one side of the main base (1) attached to it by means of a support (30).

2. The portable device according to claim 1, further characterized in that: the horizontal tubes (14) distribute the reagent located in each reagent container (12) to each of the cylindrical structures (2) through hoses (19) that are connected to a bidirectional valves (16) located inside the main base (1), preferably under each circular cavity (3).

3. The portable device according to claim 1, further characterized in that: the protrusion (32) has the same shape and dimension as the circular perforation (10) allowing the cylindrical structure (2) to be firmly held.

4. The portable device according to claim 1, further characterized in that: the connectors (15) are installed diagonally.

5. The portable device according to claim 1, further characterized in that: the vertical tube (17) has at its end a rectangular flat nozzle (34) the size of the rectangular cut (4) of the circular cavity (3) which serves to distribute the reagent to each of the slide containers (23).

6. The portable device according to claim 1, further characterized in that: in each of the bidirectional valves (16) a hose (19) is connected which is joined to each of the four horizontal tubes (14), each of which contains the reagent from each of the reagent containers (12).

7. The portable device according to claim 1, further characterized in that: on one side of the main base (1) there is a charging port (22) which serves to power the rechargeable battery (21).

8. The portable device according to claim 1, further characterized in that: the slide containers (23) are positioned vertically and have a perforation on their lower face that allows the passage of the reagent as well as the Introduction of the slides with a cytological or histological sample for staining.

9. The portable device according to claim 1, further characterized in that: the magnet (24) and the second magnet (28) have a circular shape.

10. The portable device according to claim 1, further characterized in that: the camera support (30) is of the hinge type which allows the screen to rotate on its horizontal axis.

11. The portable device according to claim 1, further characterized in that: the images acquired by the camera (8) are sent to the screen (9) and additionally to a smart device by means of the Bluetooth module (18).

Images