Automatic single cell and spatial multi-omics sample preparation and analysis system

Abstract

The invention relates to the technical field of automatic treatment of biological samples, in particular to an automatic space and single-cell multi-omics sample preparation and analysis method and system. In several examples, a continuous flexible sample strip is released from a strip dispensing assembly by a rotation mechanism. A prepared biological sample is fixed on the sample receiving surface of the flexible sample strip to generate a continuous length of prepared sample strip, and space and single cell multiomics data corresponding to the fixed biological sample is generated based on the continuous length of prepared sample strip. In some embodiments, the methods and systems are useful for preparing and analyzing solid phase biological samples, such as OCT embedded frozen tissue blocks or FFPE tissue blocks. In other embodiments, the methods and systems are suitable for preparing and analyzing liquid phase biological samples.

Description

[0001] SUMMARY: An automated single-cell and spatial multi-omics sample preparation and analysis system, comprising: a dispensing assembly for dispensing a continuous strip of flexible sample slides by a rotating mechanism; a sample preparation assembly for fixing prepared biological samples to a sample receiving surface of the flexible sample slide strip to form a continuous strip of prepared samples; and a sample analysis assembly for generating spatial and single-cell multi-omics data corresponding to the fixed biological samples based on the continuous strip of prepared samples.

[0002] In some embodiments, the prepared biological samples are solid-phase biological samples.

[0003] The sample preparation assembly comprises a first roller rotating in a first direction for receiving the flexible sample slide strip from the dispensing assembly and advancing the flexible sample slide strip towards the solid-phase biological sample so that the sample receiving surface of the flexible sample slide strip is in close contact with an attachment surface of the solid-phase biological sample; a center shaft assembly for connecting the solid-phase biological sample to a motor-controlled rotating shaft rotating in a second direction opposite to the first direction, wherein the center shaft assembly rotates in the second direction and the first roller rotates in the first direction to form a press-fit contact between the attachment surface of the solid-phase biological sample and the sample receiving surface of the flexible sample slide strip, thereby fixing the two in connection; a precision guide wheel for positioning a sectioning surface of the solid-phase biological sample close to a microtome device; and a knife blade of the microtome device for generating a tissue section block containing at least a portion of the solid-phase biological sample to form the continuous strip of prepared samples when the first roller and the center shaft assembly rotate.

[0004] In some embodiments, the solid-phase biological sample is an OCT-embedded frozen tissue block; the system further comprises a cold solution dispenser for applying a cold solution to the sample receiving surface of the flexible sample slide strip before the sample receiving surface forms a press-fit contact with the attachment surface of the solid-phase biological sample; and a cooling unit disposed near the attachment surface of the OCT-embedded frozen tissue block, wherein the cooling unit generates a cold source for synchronously cooling a local region of the OCT-embedded frozen tissue block and the applied cold solution, thereby enhancing the attachment strength between the flexible sample slide strip and the solid-phase biological sample.

[0005] In some embodiments, the solid-phase biological sample is a paraffin-embedded tissue block; the system further comprises a heating unit disposed near the attachment surface of the paraffin-embedded tissue block, wherein the heating unit generates a heat source for melting a local region of the paraffin-embedded tissue block, thereby enhancing the attachment strength between the flexible sample slide strip and the solid-phase biological sample.

[0006] The prepared biological sample is a liquid-phase biological sample. The sample preparation assembly comprises an injection controller for controlling an injector to dispense the liquid-phase biological sample onto a sample receiving surface of the flexible sample slide strip; and an application plate for applying the liquid-phase biological sample onto the sample receiving surface of the flexible sample slide strip. The sample receiving surface of the flexible sample slide strip is formed with a plurality of culture microwells for accommodating the liquid-phase biological sample containing living cells.

[0007] The sample analysis assembly comprises at least one of: a detection assembly; a multi-omics analysis assembly; and an imaging assembly.

[0008] The flexible sample slide strip is made of any one of the following materials: a thermoplastic material; a research-grade filter paper; a metal foil; or other materials that can be made into a film.

[0009] A method comprising: dispensing a continuous flexible sample slide strip by a rotating mechanism; fixing a prepared biological sample to a sample receiving surface of the flexible sample slide strip to form a continuous prepared sample strip; and generating spatial and single-cell multi-omics data corresponding to the fixed biological sample based on the continuous prepared sample strip.

[0010] The prepared biological sample is a solid-phase biological sample.

[0011] The operation of fixing the prepared biological sample to a sample receiving surface of a flexible sample slide strip to form a continuous prepared sample strip comprises: positioning the sample receiving surface of the flexible sample slide strip to a position close to an attachment surface of the solid-phase biological sample by a first roller rotating in a first direction to push the flexible sample slide strip to move towards the solid-phase biological sample; rotating a central shaft assembly of the solid-phase biological sample in a second direction opposite to the first direction to make the attachment surface of the solid-phase biological sample form a press contact with the sample receiving surface of the flexible sample slide strip, thereby achieving a fixed connection therebetween; positioning a section surface of the solid-phase biological sample to a position close to a microtome device by a precision guide wheel; and lowering a blade of the microtome device to contact the section surface while the first roller rotates in the first direction and the central shaft assembly rotates in the second direction, thereby generating a continuous tissue section sequence containing at least part of the solid-phase biological sample to form the continuous prepared sample strip.

[0012] The solid-phase biological sample is an OCT-embedded frozen tissue block, and the method further comprises: before forming the press contact between the sample receiving surface of the flexible sample slide strip and the attachment surface of the solid-phase biological sample: applying a cold solution to the sample receiving surface of the flexible sample slide strip; and after forming the press contact: emitting a cold source from a cooling unit close to the attachment surface of the OCT-embedded frozen tissue block to cool the local region of the OCT-embedded frozen tissue block and the applied cold solution, thereby enhancing the attachment strength between the flexible sample slide strip and the solid-phase biological sample.

[0013] The solid-phase biological sample is a paraffin-embedded tissue block, and the method further comprises: emitting a heat source from a heating unit close to the attachment surface of the paraffin-embedded tissue block to melt the local region of the paraffin-embedded tissue block, thereby enhancing the attachment strength between the flexible sample slide strip and the solid-phase biological sample.

[0014] The prepared biological sample is a liquid-phase biological sample. The operation of fixing the prepared biological sample to the sample receiving surface of the flexible sample slide strip to form a continuous preparation sample strip comprises: automatically dispensing the liquid-phase biological sample onto the sample receiving surface of the flexible sample slide strip by a syringe; and automatically coating the liquid-phase biological sample on the sample receiving surface of the flexible sample slide strip using a coating plate. Characterized in that: a plurality of culture microwells are formed on the sample receiving surface of the flexible sample slide strip for carrying the liquid-phase biological sample containing living cells.

[0015] A non-transitory computer-readable storage medium storing machine executable instructions that, when executed by one or more processors, cause the processors to perform the method of any one of claims 11 to 19. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 is a block diagram of an embodiment automated spatial and single-cell multi-omic sample preparation and analysis system 100; Figure 2 is a block diagram of an embodiment computing system 200 hardware architecture; Figure 3 is a top view of a sample mounting strip 105; Figure 4 is a side view of a strip dispensing assembly 110; Figure 5 is a side view of a system 100a for solid-phase sample processing; Figure 6 is a side view of a solid-phase assembly 500 in a pre-mounted state; Figure 7A is a top view of a cylindrical specimen mold 530; Figure 7B This is a schematic diagram of solid-phase biological sample 515; Figure 8 This is a side view of the slicing operation of the solid phase module 500; Figure 9 This is a top view of the prepared solid sample strip 525; Figure 10 This is a side view of the sample analysis component 130 and the strip recovery component 150; Figure 11 This is a side view of system 100b, used for liquid phase sample processing; Figure 12 This is a side view of the liquid phase assembly 600; Figure 13 This is a top view of sample slide 105, used for live cell coating; Figure 14 This is a flowchart of the solid-phase sample processing method 1400; Figure 15 This is a flowchart of the liquid phase sample processing method 1500.

[0017] Referring to the above accompanying drawings, the technical solutions of the following embodiments will now be described. These embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection thereof.

[0018] Appendix Figure 1 This is a block diagram illustrating an automated spatial and single-cell multi-omics sample preparation and analysis system 100, in which embodiments of the embodiments can be implemented. The system 100 in this embodiment is simplified for understanding; typically, the system 100 may include... Figure 1 More entities and components are shown.

[0019] In one embodiment, the space and single-cell multi-omics sample preparation and analysis system 100 may include a strip distribution component 110, a sample preparation component 120, a sample analysis component 130, a data processing module 140, and a strip retrieval component 150. In this embodiment, the strip distribution component 110 can release a continuous-length sample mounting strip 105 into the space and single-cell multi-omics sample preparation and analysis system 100, as shown in the attached diagram. Figure 4 As shown. In an embodiment, the sample mounting strip 105 may be received by a sample preparation assembly 120, in which a biological sample 115 is prepared (in one embodiment, a slice or smear) and fixed to the surface of the sample mounting strip 105, as illustrated. Figure 8 and attached Figure 12As shown. In some embodiments, the biological sample 115 may be a solid-phase biological sample, in which case the sample preparation component 120 may include an optional solid-phase component 500 configured to prepare (in one embodiment, slice) and fix the prepared biological sample onto the sample mounting strip 105. In some embodiments, the biological sample 115 may be a liquid-phase biological sample, in which case the sample preparation component 120 may include an optional liquid-phase component 600 configured to prepare (in one embodiment, smear) and fix the prepared biological sample onto the sample mounting strip 105. In embodiments, the sample mounting strip 105 on which the biological sample 115 is fixed is collectively referred to as the prepared sample strip 125. The prepared sample strip 125 may be received by the sample analysis component 130, in which the biological sample 115 is analyzed. In one embodiment, it works in conjunction with the data processing module 140, as shown in the attached figure. Figure 10 As shown. In some embodiments, the analysis of the prepared biological sample may involve one or more pre-analytical sectioning or smear preparation methods, such as removing excess OCT or paraffin cleaning, fixation, tissue clearing, pre-staining, or imaging methods, such as fluorescence imaging and / or bright-field imaging. In this case, the sample analysis component 130 may include an optional imaging component 900 or a detection component 700. In some embodiments, the analysis of the prepared biological sample may involve one or more omics characterization methods. In this case, the sample analysis component 130 may include at least one optional multi-omics characterization component 800. In embodiments, the prepared sample bands 125 that have undergone one or more analytical procedures may be received and stored by the band retrieval component 150.

[0020] Appendix Figure 2 This is a block diagram of the hardware structure of the computing system 200 in the embodiment, exemplarily illustrating a hardware configuration suitable for implementing the embodiments. The embodiments may also be implemented in other computing systems, and these systems may include components different from those described below. The computing system 200 may be configured to execute instructions for automatically preparing or analyzing space omics samples, in the embodiment implementation. In one embodiment, the computing system 200 (or multiple such systems) may be an embodiment of a strip distribution component 110, a sample preparation component 120, a sample analysis component 130, a data processing module 140, or a strip recovery component 150, or may be configured to perform the functions of these components.

[0021] Despite Figure 2Only one instance of each component is shown in the computing system 200, but there can be multiple instances of each component in the computing system 200. Also, while the computing system 200 is illustrated in the example as a single module, the system can be a single physical machine or device (in one embodiment, a single computing device implementation, such as a single workstation, a single terminal device, a single server, etc.), or can include server devices, distributed computing systems, virtual machines running on data center infrastructure, or infrastructure provided as a service (in one embodiment, virtual machines) by a cloud service provider, among other forms.

[0022] The computing system 200 includes at least one processor 202, such as a central processing unit (CPU), a microprocessor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA), a special-purpose logic circuit, a dedicated artificial intelligence processing unit, a graphics processing unit (GPU), a tensor processing unit (TPU), a neural processing unit (NPU), a hardware accelerator, or a combination of any of these processors.

[0023] The computing system 200 can include an input / output (I / O) interface 204 that enables connection to optional input devices 206 and / or optional output devices 208. In embodiments, the optional input devices 206 (in one embodiment, a keyboard, a mouse, a microphone, a camera, a scanner, a touchscreen, and / or a keypad) and the optional output devices 208 (in one embodiment, a display, a speaker, and / or a printer) are external to the computing system 200. In other embodiments, the input devices 206 and the output devices 208 can not be present, in which case the I / O interface 204 can not be needed.

[0024] The computing system 200 can include at least one communication interface 210 configured to enable wired or wireless communication with other computing systems (in one embodiment, other computing systems in a network). The communication interface 210 can include a wired connection (in one embodiment, an Ethernet cable) and / or a wireless connection (in one embodiment, at least one antenna) to enable communication within or between networks.

[0025] The computing system 200 can include at least one memory 212 (individually or collectively referred to as “memory 212”) that can be volatile or nonvolatile memory (in one embodiment, flash memory, Random Access Memory (RAM), and / or Read Only Memory (ROM)). The non-volatile memory 212 can store instructions 216 executed by the processor 202, in one embodiment, configured to implement embodiments. For example, the memory 212 can store instructions configured to perform any of the embodiment methods or embodiments. The memory 212 can also include other software instructions configured to implement an operating system (OS) and other applications or functions.

[0026] The memory 212 can also include other data 214, information, rules, policies, and machine executable instructions described herein, including, for example, spatial and single cell multi-omic data 132 obtained from the biological sample 115.

[0027] In some embodiments, the memory 212 of the computing system 200 can also include at least one electronic storage unit (not shown), in one embodiment, a solid state hard drive, a mechanical hard drive, a disk drive, and / or an optical disk drive. In certain embodiments, data and / or instructions can be provided by external memory (in one embodiment, an external drive in communication with the computing system 200 through a wired or wireless means), or by a transitory or non-transitory computer readable medium performing the functions of the memory 212.

[0028] Embodiments of the non-transitory computer readable medium include random access memory (RAM), read only memory (ROM), erasable programmable read only memory (EPROM), electrically erasable programmable read only memory (EEPROM), flash memory, compact disc read only memory (CD-ROM), or other portable memory storage. The storage unit and / or external memory can be used in conjunction with the memory 212 to implement data storage, retrieval, and caching functions for the computing system 200. The components of the computing system 200 can communicate with each other through a bus, in one embodiment.

[0029] The computing system 200 can further include a display 218, in one embodiment, a liquid crystal display (LCD) or a cathode ray tube (CRT) display, for displaying information to a user. An input device 220, in one embodiment, a keyboard and a cursor control device (such as a mouse), can be used in conjunction with the display 218. The display 218 and input device 220 can be incorporated into a touch screen in some embodiments. Figure 3is a top view diagram of a portion of a sample mounting strip 105 in an embodiment. In an embodiment, the sample mounting strip 105 can be a thin flexible strip, film or membrane made of plastic or thermoplastic material, in an embodiment, polymethyl methacrylate (PMMA), polystyrene, polycarbonate, etc., or research grade filter paper, or thin metal or metal foil (in an embodiment, metal foil) or other suitable material. The sample mounting strip 105 includes a top surface (in an embodiment, a sample receiving surface 302) and a bottom surface (not shown) configured to receive a biological sample 115. In an embodiment, the sample receiving surface 302 can receive a thin section of a solid phase biological specimen (in an embodiment, a section of a solid phase biological specimen fixed to the sample receiving surface 302 of the sample mounting strip 105 after sectioning from the solid phase biological specimen by a paraffin microtome or a cryostat microtome) or a liquid phase biological sample (in an embodiment, a smear on the sample receiving surface 302 of the sample mounting strip 105) or other forms of biological samples. In certain embodiments, to enhance the performance of diagnostic or analytical methods on the biological sample 115, the sample mounting strip 105 can be transparent, or only a portion of the sample mounting strip 105 is transparent. In an embodiment, a central region 304 of the sample mounting strip includes a predefined width 306, which can be smooth or transparent to enhance optical performance, etc. In an embodiment, the central region 304 is flanked by outer edge regions 308, which can have a coarser texture than the central region 304. In an embodiment, the coarse texture of the outer edge regions 308 can enhance the adhesion and / or cohesion between the biological sample 115 and the sample receiving surface 302 of the sample mounting strip 105 (in an embodiment, during the freeze-attachment process described below). Figure 8

[0030] In an embodiment, a series of small holes or perforations 310 can be provided on each of the outer edge regions 308 of the sample mounting strip 105 to facilitate the transport of the sample mounting strip 105 through the automated space of the single-cell multi-omics sample preparation and analysis system 100.

[0031] ​In certain embodiments, the sample mounting strip 105 can further include an encoded portion 312 located between the edge region 308 and the central region 304 of the sample mounting strip 105 that can be read by at least one glass slide position sensor for positioning as the sample mounting strip 105 is advanced in space and through the single-cell multi-omics sample preparation and analysis system 100. In certain embodiments, the encoded region can be printed or affixed on the surface of the sample mounting strip 105 at the time of manufacture, in one embodiment, on the bottom surface (not shown) of the sample mounting strip 105. In embodiments where the sample mounting strip 105 is a transparent material, the encoded portion 312 can be configured to be detected by a sensor that reads the encoded portion 312 from the top surface of the sample mounting strip 105.

[0032] FIG. 4 is an exemplary side view of a strip dispensing assembly 110 in accordance with an embodiment of the present disclosure. Figure 4 is an exemplary side view of a strip dispensing assembly 110 in accordance with an embodiment of the present disclosure. In embodiments, the strip dispensing assembly 110 is configured to release a continuous length of the flexible sample mounting strip 105 that can be loaded on a dispensing spool 402 or reel (in one embodiment, wound on the dispensing spool), by rotating the dispensing spool 402 about an axis of rotation (in one embodiment, the shaft 404) to unwind the strip. In embodiments, the dispensing spool 402 can include flanges or edges (not shown) extending from both ends thereof that are configured to help retain the sample mounting strip 105 wound on the spool. In embodiments, the dispensing spool 402 can further include a series of sprockets or protrusions (not shown) configured to engage the perforations 310 on the sample mounting strip 105 to assist in the transport of the strip 105 through the automated single-cell multi-omics sample preparation and analysis system 100. In embodiments, the center axis 406 of the dispensing spool can be mounted on a dispensing shaft 404 that is controlled by a motor, enabling the dispensing spool 402 to rotate in a first direction, and the rotational speed and direction of the shaft 404 can be controlled by the computing system 200, thereby enabling regulation of the release of the sample mounting strip 105.

[0033] In an embodiment, the sample mounting tape 105 can be fed from the dispensing assembly 110, through the sample preparation assembly 120 and the sample analysis assembly 130 in sequence, and finally collected on a collection spool of the tape collection assembly 150. In an embodiment, the sample mounting tape 105 can be extended from the dispensing spool 402 to a first roller 408 located at a predetermined distance, and then received by the sample preparation assembly 120. In an embodiment, the first roller 408 can be mounted on a first motor-controlled shaft 410, enabling the first roller 408 to rotate at a speed and direction controlled by the computing system 200. In an embodiment, the first roller 408 can also be provided with a series of sprockets or protrusions (not shown) configured to engage the perforations 310 on the sample mounting tape 105 to assist the transport of the tape 105 through the automated space of the single-cell multi-omic sample preparation and analysis system 100. In some embodiments, the first roller 408 can be equipped with an optional cleaning brush 412 configured to mechanically remove dust or debris from the surface of the first roller 408 prior to contacting the sample mounting tape 105.

[0034] In an embodiment, the first slide position sensor 414 can be mounted near the point where the sample mounting tape 105 is released from the dispensing spool 402 and configured to determine the position of the sample mounting tape 105. In one embodiment, the first slide position sensor 414 can read the coded sections 312 as the sample mounting tape 105 passes by to indicate the position on the sample mounting tape 105. In an embodiment, an optional slide coating spray or applicator unit 416 can also be mounted near the point where the sample mounting tape 105 is released from the dispensing spool 402 to spray a liquid onto the top surface (in one embodiment, the sample receiving surface 302) of the sample mounting tape 105 through a nozzle or applicator pad prior to its contact with the first roller 408. In an embodiment, the liquid can be a coating configured to more effectively bind the top surface of the sample mounting tape 105 to a biological sample. In an embodiment, the liquid can include a polylysine slide pretreatment liquid (in one embodiment, configured to improve cell culture, containing a gelatinous substance), a polylysine and water mixture, a histogel solution, and / or other coating liquids specific to the assay. In an embodiment, a gelatin solution is a commonly used slide coating adhesive that performs a function of helping to keep tissue sections attached to histological slides during staining and washing steps in an assay process. In an embodiment, the application of the liquid to the sample receiving surface 302 can also be optionally performed during the manufacturing process of the sample mounting tape 105.

[0035] Attached Figure 5is an exemplary side view drawing of an automated spatial and single-cell multi-omic sample preparation and analysis system 100a in an embodiment, configured to prepare and analyze a solid phase biological sample 515. In an embodiment, the automated spatial and single-cell multi-omic sample preparation and analysis system 100a includes a strip dispensing assembly 110 and a solid phase assembly 500, configured to prepare and mount a solid phase biological sample 515 onto a sample mounting strip 105, generating a prepared solid phase sample strip 525. In an embodiment, the automated spatial and single-cell multi-omic sample preparation and analysis system 100a further includes a sample analysis assembly 130 connected to a data processing module 140, an optional molecular processing assembly 810, and a strip recovery assembly 150. Details of the solid phase assembly 500 are described below in connection with FIGS. 5A-5C, and details of the sample analysis assembly 130 are described below in connection with FIGS. 6A-6C. This embodiment is merely illustrative of the present application and is not intended to limit the scope of the present application. Figure 6 to FIGS. 5A-5C. Figure 8 Details of the solid phase assembly 500 are described below in connection with FIGS. 5A-5C, and details of the sample analysis assembly 130 are described below in connection with FIGS. 6A-6C. This embodiment is merely illustrative of the present application and is not intended to limit the scope of the present application. Figure 10 Details of the solid phase assembly 500 are described below in connection with FIGS. 5A-5C, and details of the sample analysis assembly 130 are described below in connection with FIGS. 6A-6C. This embodiment is merely illustrative of the present application and is not intended to limit the scope of the present application.

[0036] to FIGS. 5A-5C. Figure 6 is an exemplary side view drawing of a solid phase assembly 500 in a pre-mounting state, in an embodiment, showing its cooperation with a strip dispensing assembly 110. In an embodiment, the solid phase assembly 500 is in a pre-mounting state, in one embodiment, prior to mounting a solid phase biological sample 515. In certain embodiments, the solid phase biological sample 515 can include a solid phase biological specimen embedded and frozen using an optimal cutting temperature (OCT) compound, prepared as a frozen section (in one embodiment, a cryosection), sectioned using a microtomy device such as a cryostat. In other embodiments, in one embodiment, the solid phase biological sample 515 can include a solid phase biological specimen prepared using paraffin embedding (in one embodiment, formaldehyde-fixed paraffin embedding, FFPE), for sectioning of tissue using a paraffin microtome sectioning device. In an embodiment, the melting point of paraffin configured to embed the solid phase biological sample 515 is in a range of about 56 to 62 degrees Celsius. In an embodiment, the solid phase biological specimen can be disposed in a cylindrical specimen mold 530 (FIG. 7), in one embodiment, arranged around a central axis assembly 540.

[0037] In an embodiment, the sample mounting strip 105 can be released from the dispensing spool 402, extended past the first roller 408, and positioned under the motor-controlled solid phase sample shaft 502, then extended past the second roller 508, which is positioned at a predetermined distance and vertically offset relative to the first roller 408. In an embodiment, the second roller 508 is positioned vertically lower than the first roller 408. The second roller 508 can be mounted on a second motor-controlled shaft 510, which enables the rotation of the second roller 508, and the speed and direction of the rotation are controlled by the computing system 200. The second roller 508 can also include a series of sprockets or protrusions (not shown) configured to engage the perforations 310 on the sample mounting strip 105, to assist the strip 105 in its transport through the automated space of the single-cell multi-omics sample preparation and analysis system 100a. The sample mounting strip 105 is then extended to a platform 506, which can be configured similarly to a conveyor belt, or other configurations. In an embodiment, the sample mounting strip 105 can be guided between the platform 506 and an optional rotating strip guide 512, which is configured to guide the sample mounting strip 105 along the platform 506. The strip guide 512 can be mounted on a shaft driven by a strip guide motor-controlled shaft 514, which enables the rotation of the strip guide 512, and the speed and direction of the rotation are controlled by the computing system 200. The strip guide 512 can also include a series of sprockets or protrusions 516 configured to engage the perforations 310 on the sample mounting strip 105, to assist the strip 105 in its transport through the system 100a.

[0038] In certain embodiments, the solid phase assembly 500 can also include an optional cold liquid dispensing and application device 504, which is positioned adjacent to the first roller 408, and a microtome blade holding device 518 (in an embodiment, including a microtome blade that can be mounted on the blade holding device), which can be mounted on a movable arm (not shown), details of which will be further described in the following Figure 8 In certain embodiments, the solid phase assembly 500 can also include an optional heating device 520, an optional cooling device 522, and a second coverslip position sensor 524, which is positioned below the sample mounting strip 105, details of which will be further described in the following Figure 8

[0039] ​In some embodiments, elements 504, 412, 410, 408, 520, 522, 515, and 502 may be arranged integrally on a sample advance track (not shown), allowing these elements to be advanced integrally toward the microslicing blade holder 518 during operation of the solid phase assembly 500. In one embodiment, the diameter of the solid phase sample 515 gradually decreases as it is sliced. Therefore, with every 360-degree rotation of the solid phase sample 515, elements 504, 412, 410, 408, 520, 522, 515, and 502 are advanced integrally toward the blade by a distance of 5 to 30 micrometers (i.e., the thickness of the tissue slice) to compensate for the corresponding reduction in the diameter of the solid phase biological sample 515.

[0040] Appendix Figure 7A This is a top view of the cylindrical specimen mold 530 according to an embodiment. In this embodiment, the cylindrical specimen mold 530 includes a cylindrical outer wall 532 and a circular base (not shown). The cylindrical outer wall 532 and the circular base can be easily assembled and disassembled. In one embodiment, the structure is similar to a springform pan used for baking. In this embodiment, a central shaft assembly 540 is disposed at the center of the cylindrical specimen mold 530 and configured to secure at least one solid biological specimen (in one embodiment, at least one solid biological tissue or a single solid biological tissue) within the mold 530. In this embodiment, the central shaft assembly 540 includes a plurality of protrusions 544 extending outward from the circular frame 542 into the cavity 534 of the cylindrical mold 530. These protrusions 544 may be curved or angled to achieve optimal fixation of at least one solid biological specimen, and the protrusions 544 may also have other shapes. In one embodiment, these protrusions help to secure the solid biological specimen such that, during the slicing process, when the microslicing blade applies a cutting force (in one embodiment, a downward force) to the slice surface of the solid biological sample 515 and the solid biological specimen rotates (in one embodiment, counterclockwise, or in the opposite direction to the force applied by the edge of the microslicing blade), the fixing force or frictional force applied by the protrusions 544 is greater than or equal to the cutting force, thereby firmly securing the solid biological specimen to the central axis assembly 540. In one embodiment, the central axis assembly 540 is provided with six protrusions 544, evenly distributed around the periphery of the circular frame 542, but this is only an embodiment and not limiting; other configurations may also be used.

[0041] In an embodiment, the cylindrical specimen mold 530 can be configured to prepare a solid-phase biological sample 515 (as shown in the attached document). Figure 7B(As shown). In one embodiment, after placing at least one biological tissue specimen into the cavity 534 of a cylindrical specimen mold 530, an embedding liquid (such as an OCT compound or paraffin in one embodiment) can be injected into the mold 530 to embed the specimen into a tissue block, the method including freezing or hardening the paraffin. In an embodiment, the tissue block (i.e., the solid biological sample 515) can be removed from the cylindrical specimen mold 530. In one embodiment, the embedded solid biological sample 515 is released by disassembling and separating the outer wall 532 and the base of the cylindrical specimen mold 530.

[0042] In an embodiment, the central shaft assembly 540 may include a cuboid mounting hole 546 configured to mount a solid biological sample 515 onto a motor-controlled solid sample shaft 502, enabling the solid biological sample 515 to rotate, and the rotational speed and direction of the motor-controlled shaft 502 are controlled by the computing system 200.

[0043] Appendix Figure 8 This is a side view of the solid-phase sample slicing assembly 500 according to an embodiment. In this embodiment, the solid-phase assembly 500 slices the solid-phase biological sample 515 using a continuous tissue block slicing method (in one embodiment, continuous slicing of the solid-phase biological sample 515 by rotating circular and / or spiral slicing). Specifically, the solid-phase assembly 500 receives a sample mounting strip 105 and fixes the outer surface (in one embodiment, the attachment surface) of the solid-phase biological sample 515 onto the sample mounting strip 105, while the sample mounting strip 105 is advanced along the automated space and single-cell multi-omics sample preparation and analysis system 100a. In this embodiment, after the attachment surface of the solid-phase biological sample 515 is fixed to the sample mounting strip 105, the solid-phase biological sample 515 can be sliced ​​using a microslicing blade holder 518 to obtain a series of continuous solid-phase sample slices. In one embodiment, the solid-phase biological sample 515 rotates about a motor-controlled solid-phase sample axis 502 in a second direction (e.g., opposite to the rotation direction of the first roller 408), and the micro-slicing blade tip slices its surface, thereby obtaining continuous outer surface slices. Unlike conventional solid-phase sample slicing methods that mount discrete slices on individual glass slides, this continuous tissue block slicing method allows continuous solid-phase sample slices to extend along the sample mounting strip 105 to form a continuous length. Thus, the continuous solid-phase sample slices remain fixed on the sample mounting strip 105 when output from the solid-phase assembly 500, assembling to form a prepared solid-phase sample strip 525. In some embodiments, the solid-phase assembly 500 may be installed in an adjustable cryogenic environment (in one example, a cryogenic freezer), where the solid-phase biological sample 515 is a frozen sample, and its temperature is maintained between -5 and -30 degrees Celsius.

[0044] In some embodiments, the solid phase assembly 500 can include an optional cold solution spraying and coating device 504 disposed proximate to the first roller 408. In some embodiments, when the solid phase biological sample 515 is in a frozen state (in one embodiment, previously embedded with an OCT compound and then frozen), the optional cold solution spraying and coating device 504 can spray a solution (in one embodiment, a cold solution at a temperature between 0°C and 4°C, or other solution) onto the sample receiving surface 302 of the sample mounting strip 105 to assist in securing the surface of the frozen solid phase biological sample 515 to the sample receiving surface 302. When the cold solution is applied to the sample receiving surface 302 and the sample mounting strip 105 is advanced within the solid phase assembly 500 to contact the adhering surface of the frozen solid phase biological sample 515, the optional cooling unit 522 can work in conjunction with the sample mounting strip 105 and the solid phase biological sample 515 to freeze the cold solution, causing the sample receiving surface 302 to bond or secure to the adhering surface of the solid phase biological sample 515. In embodiments, the frozen cold solution can remain frozen during the advancement of the sample mounting strip through the solid phase assembly 500, and the sample receiving surface 302 remains secured to the adhering surface of the solid phase biological sample 515 throughout the sectioning process, such that thin tissue sections taken from the solid phase biological sample 515 can remain adhered to the sample receiving surface 302. In embodiments, the thin tissue sections can be as thin as 5 microns, and can range in thickness from 30 to 50 microns. The cold solution can include molecular biology grade water or any coating solution or adhesive (in one embodiment, tissue glue), such as the Tissue-Tek® O.C.T.® Compound (Sakura Finetek USA, Inc., Torrance, CA) or other adhesive. Figure 4

[0045] In other embodiments, when the solid phase biological sample 515 is not a frozen sample, and in one embodiment, is an FFPE tissue block, another solution can be sprayed and coated onto the sample receiving surface 302 or no solution can be sprayed to assist in securing the surface of the (FFPE) solid phase biological sample 515 to the sample receiving surface 302. In one embodiment, the solution can be a glue such as tissue glue, or other adhesive can be used. In this case, the optional cooling unit 522 can not be needed, as the glue itself can adhere the adhering surface of the solid phase biological sample 515 to the sample receiving surface 302.

[0046] ​In embodiments, the sample mounting strip 105 can be advanced forward as the first roller 408 rotates in a first direction (in one embodiment, clockwise), while the central shaft assembly 540 rotates relative to the first roller 408 in a second direction (in one embodiment, counterclockwise), causing the solid phase biological sample 515 to also rotate in the second direction. This rotation causes the attachment surface of the solid phase biological sample 515 to come into compressive contact with the sample receiving surface 302 of the flexible sample strip 105, thereby causing the flexible sample strip and the solid phase biological sample to become fixedly attached. In one embodiment, the position of the surface of the first roller 408 relative to the attachment surface of the solid phase biological sample 515 is set such that a compressive force is applied to the sample receiving surface 302 and the attachment surface of the solid phase biological sample 515 as the sample mounting strip 105 passes over the first roller 408. In certain embodiments, the attached solid phase biological sample 515 and sample receiving strip 105 can pass through an optional heating unit 520. In one embodiment, when the solid phase biological sample 515 is an FFPE tissue block, the heating unit 520 can heat the FFPE tissue block, causing the paraffin embedding the biological tissue to partially melt at the point of contact with the sample receiving surface 302, ensuring that the sample receiving surface 302 is securely affixed to the surface of the FFPE tissue block. In certain embodiments, the attached solid phase biological sample 515 and sample receiving strip 105 can pass through an optional cooling unit 522. In one embodiment, when the solid phase biological sample 515 is a frozen sample, the cooling unit 522 can help maintain the desired temperature of the frozen sample. In other embodiments, in one embodiment, when the solid phase biological sample 515 is an FFPE sample, the cooling unit 522 can cause the paraffin embedding the biological tissue to re-harden, ensuring that the sample receiving surface 302 is securely affixed to the attachment surface of the FFPE sample.

[0047] In embodiments, the attached solid phase biological sample 515 and sample receiving strip 105 can pass through a second slide position sensor 524 configured to determine the position of the sample mounting strip 105. In one embodiment, the second slide position sensor 524 can read the coded portion 312 of the sample mounting strip 105 as it passes by the second slide position sensor 524 to indicate a specific position on the sample mounting strip 105.

[0048] In embodiments, a microtome blade fixation unit 518 (including a microtome blade and a blade fixation holder) can be positioned near the second roller 508 in preparation for sectioning the solid phase biological sample 515. The microtome blade can be mounted into the blade fixation holder, which is connected to a moveable arm of the microtome blade fixation unit 518, enabling the arm (and microtome blade) to move up and down automatically. The microtome blade can be moved from a holding position to a sectioning position, in contact with the solid phase biological sample 515. As the surface of the solid phase biological sample 515 rotates past the tip of the microtome blade, successive sections of the solid phase biological sample can be obtained. In embodiments, the second roller 508 can act as a precision sectioning guide wheel, guiding the microtome blade fixation unit 518 to precisely cut the solid phase biological sample 515. The successive sections will be affixed to the sample receiving surface 302 of the sample mounting strip 105, forming a prepared solid phase sample strip 525. The solid phase sample strip 525 will travel along a path through the solid phase assembly 500, and contact the guide 512 at the platform 506, before being received by the sample analysis assembly 130. The path design creates a gap 526 between the second roller 508 and the guide 512, configured to reduce tangential compressive and / or tensile stress applied to the solid phase sample strip 525, ensuring that the successive sections are securely affixed to the receiving surface 302 of the sample mounting strip 105. In certain embodiments, the solid phase sample strip 525 is oriented in a substantially vertical orientation near the microtome blade fixation unit 518, and transitions to a substantially horizontal orientation near the platform 506, and the gap 526 facilitates this gentle transition.

[0049] attached Figure 9 A top view of a portion of the prepared solid phase sample strip 525 is shown in embodiments. In embodiments, the solid phase sample strip 525 includes successive solid phase biological sample sections 902 affixed to the sample receiving surface 302, extending along a predefined width 906. The successive solid phase biological sample sections 902 are generally located within the central region 304 of the sample mounting strip, such that the coded region 312 located between the edge region 308 and the central region 304 can be read by at least one sample strip position sensor 904. In some embodiments, other sensors or scanners (in one embodiment, the imaging device scanner 706 described in the Methods section) can also be arranged to scan or analyze the successive solid phase biological sample sections 902. Figure 10

[0050] In embodiments, the sample mounting strip 105 is provided with perforations 310 along each of the outer side edge regions 308, which can be engaged by sprockets or protrusions 516 on the at least one strip guide 512, thereby assisting in the smooth transport of the sample mounting strip in space and single cell multi-omics sample preparation and analysis system 100a.

[0051] attached Figure 10 ​A side view of the sample analysis assembly 130 and the strip collection assembly 150 is shown. In embodiments, the sample analysis assembly 130 can include a detection assembly 700, a multi-omic analysis assembly 800, and an imaging assembly 900. In some embodiments, the prepared solid-phase sample strip 525 can be transported through the sample analysis assembly 130 configured to analyze the sequenced solid-phase biological sample sections 902 to obtain spatial and single-cell multi-omic data 132, in one embodiment, delivered on the platform 506 (in one embodiment, along the platform 506 with the assistance of at least one strip guide 512) by various sensors and / or technologies. In other embodiments, the prepared liquid-phase sample strip 625 (as described in the Figure 12 above) can also be delivered through the sample analysis assembly 130 configured to analyze the liquid-phase biological sample 615 to obtain spatial and single-cell multi-omic data 132 in a similar manner. It is noted that while the assemblies 700, 800, and 900 are described in a particular order, they can be arranged in other orders, and multiple identical assemblies can be used or portions thereof can be omitted.

[0052] In one embodiment, the imaging assembly 700 can include a heated blow-dry unit 702 configured to prepare the sequenced solid-phase biological sample sections 902 for analysis. The heated blow-dry unit 702 can blow a warm air stream to the solid-phase biological sample sections 902 to help dry and fix the sample. The imaging assembly 700 can also include at least one slide position sensor 904 configured to read the position of the prepared solid-phase sample strip 525. In addition, the imaging assembly 700 can be equipped with at least one imaging device scanner 706. A vacuum device 708 can be provided at the bottom of the platform 506 near the imaging device scanner 706 to create a suction force by drawing air through at least one opening 710 (in one embodiment, a row of pinholes, a set of pinholes, or a slit, etc.) on the platform 506. The suction force helps to ensure that the prepared solid-phase sample strip 525 can be flatly attached to the platform 506 when scanned by the imaging device scanner 706. The imaging data (in one embodiment, the spatial and single-cell multi-omic data 132) obtained by the imaging device scanner 706 can be transmitted to the data processing module 140 for analysis.

[0053] In one embodiment, the multi-omics analysis assembly 800 can include a molecule extraction assembly configured to extract DNA, RNA, proteins, etc. molecules from the sequenced solid-phase biological sample slice 902. In embodiments, the multi-omics analysis assembly 800 can also include a molecule sample fixation strip dispensing assembly configured to continuously dispense a length of flexible molecule sample fixation strip 805 from a dispensing spool 802 (in one embodiment, a spool wound around a rotating shaft) by rotating the dispensing spool 802 about an axis of rotation (in one embodiment, a bearing) in a first direction. The rotation speed and direction of the dispensing spool 802 can be controlled by the computing system 200. The molecule sample fixation strip 805 can be pre-coated with a barcoded capture molecule such as an oligonucleotide, an antibody, or other chemical substance on its fixation surface. The dispensing spool 802 can be provided with flanges or edges (not shown) around its two ends to help secure the molecule sample fixation strip 805; it can also be provided with a series of gears or protrusions (not shown) configured to engage perforations on the molecule sample fixation strip 805 to assist in pushing the molecule sample fixation strip 805 through the multi-omics analysis assembly 800.

[0054] In embodiments, the molecule sample fixation strip 805 can extend from the dispensing spool 802 and pass through or around a roller 806 mounted at a predetermined distance and another roller 808 mounted at a predetermined distance from the roller 806. The rollers 806 and 808 are mounted on motor shafts controlled by the computing system 200, allowing the rollers to rotate, and the rotation speed and direction of the rollers can be controlled by the computing system 200. The rotation of the rollers causes the molecule fixation surface of the molecule sample fixation strip 805 to come into compressive contact with the prepared solid-phase sample strip 525, thereby effecting extraction or transfer of biological contents from the biological sample on the solid-phase sample strip 525 onto the molecule sample fixation strip 805. The rollers 806 and / or 808 can include a series of gears or protrusions (not shown) configured to engage perforations on the sample fixation strip 105 or the molecule sample fixation strip 805 to assist in pushing these two sample fixation strips through the multi-omics analysis assembly 800.

[0055] In embodiments, the coating spray or application device 804 can be mounted near where the molecule sample fixation strip 805 is released from the dispensing spool 802 and configured to spray or apply a liquid through a nozzle or an applicator plate onto the molecule sample fixation strip 805 before the molecule sample fixation strip 805 contacts the roller 806. The sprayed liquid can be a reagent or a biochemical substance or solution configured for analysis.

[0056] Additional rollers (not shown) can further guide the molecular sample holder 805 through the multi-omics analysis component 800, and the corresponding extracted molecular samples can be transferred to the molecular processing component 810 for further analysis (in one embodiment, DNA / RNA sequencing or other molecular imaging procedures). In embodiments, molecular data obtained from the molecular processing component 810 (in one embodiment, spatial and single-cell multi-omics data 132) can also be transferred to the data processing module 140 for analysis.

[0057] In this embodiment, the data processing module 140 can integrate data and perform three-dimensional reconstruction on the solid biological sample 515. In one embodiment, it integrates data from the slice position sensor with multi-omics imaging and sequencing data, and reconstructs a three-dimensional model of the cellular and molecular composition of the biological tissue or organ. Taking the pancreas as an example, its dimensions are approximately 15 cm × 7 cm × 4 cm (corresponding to a volume of approximately 420 cubic centimeters), and it can be formed into different shapes in a cylindrical mold 530 before slicing. However, the tissue sample after slicing will be arranged in a two-dimensional plane during detection. At this time, data processing is required to arrange the slices according to the curvature, thereby reconstructing the corresponding three-dimensional digital pancreas. Although such reconstruction can be achieved by CT or MRI instruments (such as equipment in hospital settings), their resolution is usually low. For liquid samples, the data processing module 140 mainly collects and analyzes the cellular and molecular information of the sample.

[0058] In this embodiment, although the sample analysis component 130 is described within the context of an automated spatial and single-cell multi-omics sample preparation and analysis system 100a (specifically configured for the preparation and analysis of solid-phase biological samples 515), it should be understood that this embodiment is not a limiting description. According to other embodiments, the sample analysis component 130 can also be adapted to other embodiments of the automated spatial omics sample preparation and analysis system 100a, in one embodiment, specifically configured as a system for the preparation and analysis of liquid-phase biological samples or other forms of biological samples.

[0059] Appendix Figure 11 The figure shown is an exemplary side view of an automated spatial and single-cell multi-omics sample preparation and analysis system 100b, specifically configured for the preparation and analysis of liquid biological samples 615. In this embodiment, system 100b includes: a strip dispensing assembly 110, a liquid phase assembly 600 (configured to prepare and load the liquid biological sample 615 onto a sample slide strip 105 to form a prepared liquid sample strip 625), and system 100b also integrates: a sample analysis assembly 130 connected to a data processing module 140, an optional molecular processing assembly 810, and a strip recovery assembly 150. A detailed description of the liquid phase assembly 600 will be provided below in conjunction with the accompanying drawings. Figure 12 This embodiment is only used to illustrate the technical solution of the present invention, and is not intended to limit its scope of protection.

[0060] Appendix Figure 12 The accompanying drawing shows a side view of an exemplary embodiment of the liquid sample coating assembly 600. In some embodiments, the liquid biological sample 615 may contain a specific concentration of a biological tissue cell suspension. This concentration setting must ensure that the biological tissue cells are uniformly distributed in a monolayer or sub-monolayer (other distribution methods may also be used). Specifically, the liquid biological sample 615 may be loaded into a disposable syringe 602 and then dispensed onto a sample slide strip 105, configured to perform single-cell smear flow cytometry—a technique that enables the joint analysis of single-cell surface markers with multi-omics data (and other possible analytical methods).

[0061] In an embodiment, the sample carrier strip 105 can be obtained from the attached... Figure 4 The sample strip is unfurled on the distribution reel 402. The strip is then conveyed to the liquid phase assembly 600—specifically, the sample strip 105 extends to platform 506 and is received by an optional strip guide 612, which guides the strip along platform 506. The guide 612 is mounted on a motor-controlled guide shaft 614, the speed and direction of which are controlled by the computing system 200. In particular, the guide 612 surface is provided with a series of teeth / protrusions 516 configured to engage the perforations 310 of the sample strip 105, thereby ensuring precise delivery of the strip in space and within the single-cell multi-omics sample preparation and analysis system 100b.

[0062] In some embodiments, the liquid biological sample 615 can be dispensed onto the sample slide 105 at a preset rate via the injection controller 604. In one embodiment, the injection controller 604 can cooperate with the syringe 602 to automatically and continuously dispense a quantitative amount of liquid biological sample 615 from the nozzle 606 onto the sample slide 105 at a preset rate based on parameters such as the conveying speed of the sample slide 105 on the platform 506 or the diameter of the nozzle 606. The dispensing speed can be controlled by the computing system 200. In some embodiments, the nozzle 606 can be connected to the syringe 602 via a connecting tube 608.

[0063] In this embodiment, after the liquid biological sample 615 is dispensed onto the sample slide strip 105, it can be uniformly coated onto the slide strip using a coating plate 610. The coating plate 610 can be made of glass, stainless steel, or other suitable materials, and its width is designed to be equal to or less than the preset width 306 of the central region 304 of the sample slide strip 105. The coating plate 610 is fixed to the plate holder 616 and positioned relative to the platform 506 at a preset distance and preset angle to ensure that after contacting the liquid biological sample 615 on the sample slide strip 105, it can uniformly coat the sample with a predetermined thickness onto the sample receiving surface 302 of the sample slide strip 105, thereby forming the prepared liquid sample strip 625.

[0064] In some embodiments, a vacuum device 620 may be positioned below the platform 506 near the coating plate 610. This device generates suction by drawing air through at least one opening 622 on the platform 506 (in one embodiment, an array of small holes, a grid of small holes, or slits, etc.). This vacuum suction helps ensure that the prepared liquid sample strip 625 adheres sufficiently flat to the platform 506 during the coating process. Subsequently, the prepared liquid sample strip 625 can be transferred to the sample analysis assembly 130 for testing; the specific procedure can be found in the appendix above. Figure 10 As stated above.

[0065] Appendix Figure 13 A top view of an embodiment of a sample slide strip 105 suitable for live cell suspension coating and in vivo culture detection and analysis is shown as an example. The slide strip is a flexible thin strip made of plastic or other suitable material, comprising a plurality of micropores 320 formed on its upper surface (i.e., the sample receiving surface 302), configured to carry a liquid biological sample 615. This liquid sample is typically a biological tissue cell suspension of a specific concentration, the concentration of which must be set to ensure uniform distribution of biological tissue cells in each micropore 320. As described above... Figure 12 As shown, a quantitative liquid biological sample 615 can be dispensed onto the sample receiving surface 302 using a syringe 602, and then coated using a coating plate 610, ultimately forming a prepared liquid sample strip 625 in each microwell 320.

[0066] In some embodiments, the sample slide 105 may be made of a transparent material or have a partially transparent design to improve the diagnostic performance of the liquid biological sample 615. In one embodiment, the central region 304 of the slide may include a smooth or transparent surface of a predetermined width 306 to optimize optical detection characteristics. The central region 304 is flanked by edge regions 308, whose surface texture is rougher than that of the central region 304.

[0067] In some embodiments, the sample slide strip 105 can be provided with a series of small holes or perforations 310 along the two side edge regions 308 for facilitating the transport of the slide strip in the automated spatial and single-cell multi-omic sample preparation and analysis system 100. In addition, the edge regions can be designed with a rough surface texture to assist in the stable transport of the sample slide strip in the spatial and single-cell multi-omic sample preparation and analysis system 100 by increasing the frictional force with the chain-and-sprocket components such as the strip dispensing assembly 110 and the strip recovery assembly 150.

[0068] In some embodiments, the sample slide strip 105 can also be provided with a coded region 312 between the edge regions 308 and the central region 304. The coded region can be read by at least one slice position sensor during the transport of the slide strip. The coded region can be printed or affixed to the surface of the slide strip (in one embodiment, the bottom surface of the slide strip) at the time of manufacture, and the coded information can still be read by a sensor positioned above the slide strip when a transparent material is used.

[0069] Figure 1 is a schematic diagram of an embodiment of an automated spatial and single-cell multi-omic sample preparation and analysis system 100. The system 100 is suitable for the preparation and analysis of solid-phase biological samples such as OCT-embedded frozen tissue blocks or FFPE tissue blocks. Figure 14 Figure 1 is a schematic diagram of an embodiment of an automated spatial and single-cell multi-omic sample preparation and analysis system 100. The system 100 is suitable for the preparation and analysis of solid-phase biological samples such as OCT-embedded frozen tissue blocks or FFPE tissue blocks.

[0070] The method 1400 begins at step 1402 by dispensing a continuous flexible sample slide strip 105 from the strip dispensing assembly 110 by a rotating mechanism. The continuous flexible sample slide strip 105 can be pre-loaded on a dispensing spool 402, and the release of the slide strip can be achieved by rotating the spool around an axis of rotation such as the shaft 404. As the slide strip is released from the dispensing assembly, the nozzles of the spray-on coating unit 416 can spray a liquid onto the sample receiving surface 302 of the slide strip. The liquid coating can enhance the bonding effect of the sample receiving surface with the adhering surface of the solid-phase biological sample.

[0071] At step 1404, the prepared biological sample 115 is affixed to the sample receiving surface 302 of the flexible sample slide strip 105 to form a continuous prepared sample strip 125. In some embodiments, the prepared biological sample 115 can be a solid-phase biological sample 515, and the preparation process can include placing the biological specimen affixed to the center shaft assembly 540 together with cold OCT or hot molten paraffin in a cylindrical specimen mold 530, allowing the contents to harden (e.g., cooling the paraffin-embedded tissue to room temperature or lowering the frozen sample to below -10 degrees Celsius), and then mounting the prepared biological sample 515 on the solid-phase sample shaft 502 of the solid-phase assembly 500. The specific steps for affixing the prepared biological sample 515 to the sample receiving surface 302 of the flexible sample slide strip 105 are as follows.

[0072] In step 1406, the sample receiving surface 302 of the flexible sample carrier strip 105 is positioned near the attachment surface of the solid biological sample 515 by the first roller 408. When the first roller 408 rotates in the first direction, it can push the flexible sample carrier strip 105 toward the solid biological sample 515.

[0073] In step 1408, the central axis assembly 540 of the solid biological sample 515 is rotated in a second direction opposite to the first direction, causing the attachment surface of the sample to form a pressurized contact with the sample receiving surface 302 of the flexible sample carrier strip 105. This pressurization action achieves a stable bond between the flexible sample carrier strip 105 and the solid biological sample 515.

[0074] In step 1410, the surface of the solid biological sample 515 is positioned close to the microslicing device (such as the microtome clamping unit 518) using a second roller 508 (such as a precision guide roller).

[0075] In step 1412, as the first roller 408 rotates in the first direction and the central axis assembly 540 of the solid biological sample 515 rotates in the second direction, the blade of the microslicing device presses down to contact the sample slice surface, thereby generating a continuous sequence of tissue slices containing portions of the solid biological sample. These slices are firmly attached to the flexible sample carrier strip, forming a continuous sample preparation strip 125.

[0076] In step 1414, spatial and single-cell multi-omics data 132 corresponding to the fixed biological sample are generated based on the continuous preparation of sample strips 125. The specific process includes: the preparation of sample strips 125 first undergoes tissue pretreatment (such as preserving tissue state using methods like heating, freezing, chemical fixation, or enzymatic treatment) through at least one detection component 700, and then is transported to an imaging scanner 706 for sample scanning to generate attached image data. The sample strips will also pass through a molecular extraction component to extract molecules such as DNA, RNA, and proteins from the sample. The extracted molecular samples will be sent to a molecular processing component 810 for in-depth analysis. Finally, the system will integrate and analyze the spatial and single-cell multi-omics data 132 from the imaging scanner 706 and the molecular processing component 810.

[0077] Appendix Figure 15 An exemplary flowchart of an embodiment method 1500 configured for automated spatial and single-cell multi-omics sample preparation and analysis is shown. This method is particularly suitable for configuration for the preparation and analysis of liquid-phase biological samples.

[0078] Method 1500 begins at step 1502, in which a continuous flexible sample carrier strip 105 is released from the strip dispensing assembly 110 via a rotation mechanism. The carrier strip is typically pre-loaded onto a dispensing reel 402, and its unfolding and release are achieved by rotating the reel about a rotation axis (such as axis 404).

[0079] In step 1504, the prepared biological sample 115 is fixed onto the sample receiving surface 302 of the flexible sample slide strip 105, forming a continuous prepared sample strip 125. When the sample is a liquid phase biological sample 615, the fixing process includes the following steps.

[0080] In step 1506, the liquid phase biological sample 615 is automatically dispensed onto the sample receiving surface 302 of the flexible sample slide strip 105 by the syringe 602.

[0081] In step 1508, the liquid phase biological sample 615 is automatically coated on the sample receiving surface 302 of the flexible sample slide strip 105 using the coating plate 610.

[0082] In step 1510, spatial and single-cell multi-omics data 132 corresponding to the fixed biological sample is generated based on the continuous prepared sample strip 125. The specific process includes: (1) the sample strip 125 is subjected to tissue pretreatment (such as heat fixation, freezing, chemical fixation, or enzyme treatment, etc.) by the detection assembly 700; (2) the image data is generated by scanning the imaging scanner 706; (3) biological molecules such as DNA / RNA / proteins are extracted by the molecular extraction assembly; (4) the extract is sent to the molecular processing assembly 810 for in-depth analysis. Finally, the system integrates the multi-omics data 132 from the imaging scanner and the molecular assembly for comprehensive analysis.

[0083] Although the above embodiments describe the method process including a specific sequence of steps, in actual application, some steps can be omitted or the execution order of some steps can be adjusted according to specific circumstances. The execution order of each step is not necessarily strictly limited to the order described in the specification. The embodiments are only used to illustrate the technical solutions of the present application, and not to limit the protection scope thereof.

[0084] Although the present specification describes the embodiments by method processes, those skilled in the art should understand that these embodiments also relate to various assemblies (including hardware assemblies, software, or a combination thereof) configured to implement at least part of the functions in the described methods. The technical solutions of the exemplary embodiments can be embodied in the form of a software product, which can be stored in a pre-recorded non-volatile storage medium such as a DVD, CD-ROM, U disk, mobile hard disk, etc. The software product contains executable instructions that can make an electronic device (such as a personal computer, a server, or a network device) implement the method embodiments described in the present specification. The embodiments are only used to illustrate the technical solutions of the present application, and not to limit the protection scope thereof.

[0085] In the methods, systems, devices, or flowcharts described herein, each functional block can represent an event, step, function, flow, module, signal, and / or state operation, etc. Although some embodiments describe a specific order of execution, the actual order of execution can differ from that described. Additionally, certain steps / states can be omitted or combined, and other steps / states can be inserted, in different embodiments. Furthermore, some steps / states can be performed concurrently, in different embodiments. Some or all of the steps / states can be performed by a hardware, software, or firmware component. The steps / states can be repeated any number of times. The steps / states can also be configured as a system or sub-components thereof, and vice versa. For "send," "receive," and the like, the perspective of the device can be interchanged.

[0086] The technical solutions disclosed in the present specification can be implemented in other specific forms without departing from the core idea of the embodiments. The embodiments should be understood in an exemplary and non-limiting manner. Those skilled in the art can combine features in at least one embodiment to form alternative embodiments not explicitly described, and such combinations of features should be considered to be within the scope of the embodiments.

[0087] The numerical ranges disclosed in the present specification include all sub-ranges and specific numerical values within the ranges. Although the described systems and devices include a specific number of components, the number of components can be increased or decreased (e.g., a single component can be expanded to multiple components) in actual implementation. The technical solutions described herein are intended to cover all reasonable technical variations.

Claims

1. An automated single-cell and spatial multi-omics sample preparation and analysis system, comprising: a dispensing assembly for dispensing a continuous flexible sample slide strip by a rotating mechanism; a sample preparation assembly for fixing a prepared biological sample to a sample receiving surface of the flexible sample slide strip to form a continuous prepared sample strip; and a sample analysis assembly for generating spatial and single-cell multi-omics data corresponding to the fixed biological sample based on the continuous prepared sample strip. The prepared biological sample is a solid-phase biological sample.

2. The system of claim 1, wherein, The sample preparation assembly comprises 3. The system of claim 2, wherein, a first roller rotating in a first direction for receiving the flexible sample slide strip from the dispensing assembly and advancing the flexible sample slide strip towards the solid-phase biological sample such that the sample receiving surface of the flexible sample slide strip is in close contact with an attachment surface of the solid-phase biological sample; a center shaft assembly for connecting the solid-phase biological sample to a motor-controlled rotating shaft rotating in a second direction opposite to the first direction, wherein the center shaft assembly rotates in the second direction and the first roller rotates in the first direction to form a press-fit contact between the attachment surface of the solid-phase biological sample and the sample receiving surface of the flexible sample slide strip, thereby fixing the two together; a precision guide wheel for positioning a sectioning surface of the solid-phase biological sample close to a microtome device; and a knife of the microtome device for generating a tissue section block containing at least a portion of the solid-phase biological sample to form the continuous prepared sample strip when the first roller and the center shaft assembly rotate.

4. The system of claim 3, wherein: the solid-phase biological sample is an OCT-embedded frozen tissue block; and the system further comprises: a cold solution dispenser for applying a cold solution to the sample receiving surface of the flexible sample slide strip before the sample receiving surface forms the press-fit contact with the attachment surface of the solid-phase biological sample; and a cooling unit disposed adjacent to the attachment surface of the OCT-embedded frozen tissue block, the cooling unit emitting a cold source for synchronously cooling a local region of the OCT-embedded frozen tissue block and the applied cold solution, thereby enhancing an attachment strength between the flexible sample slide strip and the solid-phase biological sample.

5. The system of claim 3, wherein: the solid-phase biological sample is a paraffin-embedded tissue block; and the system further comprises: a heating unit disposed adjacent to the attachment surface of the paraffin-embedded tissue block, the heating unit emitting a heat source for melting a local region of the paraffin-embedded tissue block, thereby enhancing an attachment strength between the flexible sample slide strip and the solid-phase biological sample. The prepared biological sample is a liquid-phase biological sample. The sample preparation assembly comprises: an injection controller for controlling an injector to dispense the liquid-phase biological sample onto the sample receiving surface of the flexible sample slide strip; and 6. The system of claim 1, wherein, a spreading plate for spreading the liquid-phase biological sample on the sample receiving surface of the flexible sample slide strip.

7. The system of claim 6, wherein, 8. The system of claim 6, wherein: ​ ​ ​ A plurality of culture micro-wells are formed on the sample receiving surface of the flexible sample slide strip for receiving a liquid phase biological sample containing living cells.

9. The system of claim 1, wherein, The sample analysis assembly comprises at least one of: a detection assembly; a multi-omics analysis assembly; and 10. The system of claim 1, wherein, an imaging assembly. The flexible sample slide strip is made of any one of the following materials: a thermoplastic material; a research grade filter paper; a metal foil; or other materials that can be made into a thin film.

11. A method comprising: dispensing a continuous flexible sample slide strip by a rotating mechanism; fixing a prepared biological sample to a sample receiving surface of the flexible sample slide strip to form a continuous prepared sample strip; and generating spatial and single-cell multi-omics data corresponding to the fixed biological sample based on the continuous prepared sample strip.

12. The method of claim 11, wherein, The prepared biological sample is a solid phase biological sample.

13. The method of claim 12, wherein, The operation of fixing the prepared biological sample to the sample receiving surface of the flexible sample slide strip to form a continuous prepared sample strip comprises: positioning the sample receiving surface of the flexible sample slide strip to a position close to an attachment surface of the solid phase biological sample by a first roller rotating in a first direction to push the flexible sample slide strip to move towards the solid phase biological sample; rotating a central shaft assembly of the solid phase biological sample in a second direction opposite to the first direction to make the attachment surface of the solid phase biological sample form a press contact with the sample receiving surface of the flexible sample slide strip, thereby achieving a fixed connection therebetween; positioning a section surface of the solid phase biological sample to a position close to a microtome device by a precision guide wheel; and lowering a blade of the microtome device to contact the section surface while the first roller rotates in the first direction and the central shaft assembly rotates in the second direction, thereby generating a continuous tissue section sequence containing at least part of the solid phase biological sample, forming the continuous prepared sample strip.

14. The method of claim 13, wherein, The solid phase biological sample is an OCT-embedded frozen tissue block, and the method further comprises: before making the sample receiving surface of the flexible sample slide strip form a press contact with the attachment surface of the solid phase biological sample: applying a cold solution to the sample receiving surface of the flexible sample slide strip; and after forming the press contact: emitting a cold source from a cooling unit close to the attachment surface of the OCT-embedded frozen tissue block to cool a local area of the OCT-embedded frozen tissue block and the applied cold solution, thereby enhancing the attachment strength between the flexible sample slide strip and the solid phase biological sample.

15. The method of claim 13, wherein, The solid phase biological sample is a paraffin-embedded tissue block, and the method further comprises: emitting a heat source from a heating unit close to the attachment surface of the paraffin-embedded tissue block to melt a local area of the paraffin-embedded tissue block, thereby enhancing the attachment strength between the flexible sample slide strip and the solid phase biological sample.

16. The method of claim 11, wherein, The prepared biological sample is a liquid phase biological sample.

17. The method of claim 16, wherein, The operation of fixing the prepared biological sample to the sample receiving surface of the flexible sample slide strip to form a continuous prepared sample strip comprises: automatically dispensing the liquid phase biological sample onto the sample receiving surface of the flexible sample slide strip by a syringe; and The liquid-phase biological sample is automatically coated on the sample receiving surface of the flexible sample slide strip using a coating plate.

18. The method of claim 16, wherein: A plurality of culture microwells are formed on the sample receiving surface of the flexible sample slide strip for carrying the liquid-phase biological sample containing living cells.

19. The method of claim 1, wherein, The flexible sample slide strip is made of any of the following materials: thermoplastic material; filter paper of research grade; metal foil; or other material that can be made into a thin film.

20. A non-transitory computer-readable storage medium storing machine executable instructions that, when executed by one or more processors, cause the processors to perform the method of any one of claims 11 to 19.

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