CN-121975922-A - Method and system for releasing space histology chip probe based on image guidance

CN121975922ACN 121975922 ACN121975922 ACN 121975922ACN-121975922-A

Abstract

The application discloses a method and a system for releasing a space histology chip probe based on image guidance, which relate to the technical field of space transcriptomics, and the method comprises the steps of shooting by a camera to obtain an original image of a tissue slice; the tissue slice is attached to the surface of a probe coding chip, a probe incubation reagent is added to the tissue slice, an original image is preprocessed, a tissue region in the original image is obtained through positioning, the tissue region is used as a tissue mask, the tissue mask is converted from camera coordinates to digital micro-mirror device coordinates, a control signal of the digital micro-mirror device is generated according to the digital micro-mirror device coordinates, and light generated by a light source is projected to the tissue region by the digital micro-mirror device in response to the control signal so as to release a corresponding probe in the tissue region range in the probe coding chip. According to the application, the probe coding chip is irradiated according to the customized irradiation pattern, so that only the probe in the range of the tissue area is released, and the precise illumination control of the tissue area is realized.

Inventors

WANG BING
WANG ZHIFENG
LUO XIONGJIAN
He Zhuoying
YANG ZHIBIN

Assignees

利德健康科技(广州)有限公司

Dates

Publication Date: 20260505
Application Date: 20251218

Claims (10)

1. The method for releasing the space histology chip probe based on image guidance is characterized by comprising the following steps of: shooting an original image of a tissue slice by using a camera, wherein the tissue slice is attached to the surface of a probe coding chip; Adding a probe incubation reagent to the tissue section; preprocessing the original image so as to locate and obtain a tissue region in the original image; Converting the tissue region as a tissue mask from camera coordinates to digital micromirror device coordinates; generating a control signal of the digital micro-mirror device according to the coordinates of the digital micro-mirror device; and responding to the control signal, and utilizing the digital micro-mirror device to project light generated by a light source to the tissue region so as to release the corresponding probes in the tissue region range in the probe coding chip.
2. The method for releasing the image-guided spatial histology chip probe according to claim 1, wherein the preprocessing of the original image to locate the tissue region in the original image comprises the following steps: identifying tissue contours in the original image using threshold segmentation, a deep learning model, or edge detection and morphological processing; Binarizing the original image according to the tissue outline to distinguish and obtain a tissue area and a non-tissue area in the original image; And correcting the edge of the tissue region by using morphological opening and closing operation so as to enable the mask corresponding to the tissue region to be continuous and smooth in illumination boundary.
3. The method for releasing the spatial histology chip probe based on image guidance according to claim 1, wherein the generating the control signal of the digital micro-mirror device according to the coordinates of the digital micro-mirror device comprises the steps of: distributing switching instructions of the micromirror array according to different assignment of coordinates of the digital micromirror device; Outputting the switch instruction to a micromirror state matrix; Converting the micromirror state matrix into a binary image frame, writing the binary image frame into a display buffer of a controller of the digital micromirror device, and starting a single-frame static projection mode; And generating the control signal synchronously triggered by the light source and the digital micro-mirror device.
4. The method of image guided spatially diverse chip probe release of claim 1, further comprising the steps of: Selecting the light source matched with the characteristics of the light fracture connection sub, wherein the wavelength of the light source comprises an ultraviolet band, a visible light band or a near infrared band; and a shading component or a scattering suppression module is arranged in the light path system of the light source so as to reduce the influence of stray light or reflected light.
5. The method of claim 1, wherein the projecting light generated by a light source onto the tissue region using the digital micromirror device in response to the control signal to release the corresponding probe in the probe-encoded chip within the tissue region comprises the steps of: Switching an inclined state by using a micromirror array on the digital micromirror device according to the control signal in response to the control signal to realize spatial modulation of an optical signal, wherein the inclined state comprises on and off; When the tilting state of the micromirror array is on, controlling incident light to reflect to a projection light path, so that the modulated light pattern of the tissue mask irradiates the tissue region in the tissue slice; when the tilt state of the micromirror array is off, the incident light is controlled to deflect to the light absorbing assembly or light trap.
6. The method of claim 5, wherein controlling reflection of incident light to a projection light path to irradiate the modulated light pattern of the tissue mask to the tissue region in the tissue slice when the tilt state of the micromirror array is on comprises: regulating the light source to output light with corresponding wavelength according to the tissue region by utilizing a control module according to the control signal, so that the modulated light pattern of the tissue mask irradiates the tissue region in the tissue slice; The wavelength and the power of the light output by the light source are selected according to the characteristics of the light fracture connector connected to the probe coding chip; The irradiation time was determined based on release efficiency, probe density and photoreaction kinetic parameters.
7. The method of image guided spatially diverse chip probe release of any one of claims 1 to 6, further comprising periodically performing an alignment verification step comprising the steps of: before an experiment, a control module is utilized to enable the digital micro-mirror device to project a preset calibration pattern to the plane of a tissue slice sample, and a tissue contour image of the tissue slice sample is recorded in advance; when the alignment checking step is executed, the digital micro-mirror device is used for projecting a preset calibration pattern to the plane of the tissue slice sample by using the control module; Collecting the projected pattern image by using an imaging device; Comparing the pattern image with a previously recorded tissue contour image, and calculating spatial deviation information through an image registration algorithm, wherein the spatial deviation information comprises translational errors, rotation angle deviations, inclination angles and nonlinear distortion parameters; And driving a platform or an optical component to perform automatic fine adjustment by utilizing the control module according to the space deviation information, wherein the automatic fine adjustment comprises at least one of moving a sample platform, adjusting a projection lens group and correcting the angle or focal length of a panel of the digital micro-mirror device.
8. An image-guided, spatially-componential chip probe release system, the system comprising: The tissue slice processing device comprises an image shooting unit, a probe coding chip, a probe incubating reagent and a probe detecting unit, wherein the image shooting unit is used for shooting an original image of a tissue slice by using a camera, and the tissue slice is attached to the surface of the probe coding chip; The tissue positioning unit is used for preprocessing the original image so as to position and obtain a tissue region in the original image; a coordinate conversion unit for converting the tissue mask from camera coordinates to digital micromirror device coordinates with the tissue region as a tissue mask; a signal generating unit, configured to generate a control signal of the digital micromirror device according to the coordinates of the digital micromirror device; and the irradiation control unit is used for responding to the control signal, and utilizing the digital micro-mirror device to project the light generated by the light source to the tissue area so as to release the corresponding probes in the tissue area range in the probe coding chip.
9. An electronic device comprising a memory storing a computer program and a processor implementing the method of any of claims 1 to 7 when the computer program is executed by the processor.
10. A computer readable storage medium storing a computer program, characterized in that the computer program when executed by a processor implements the method according to any one of claims 1 to 7.

Description

Method and system for releasing space histology chip probe based on image guidance Technical Field The application relates to the technical field of space transcriptomics, in particular to a method and a system for releasing a space transcriptomics chip probe based on image guidance. Background In the study of space histology, especially space transcriptomics, mainly relies on space histology chips with coded probes, which are required to capture RNA or release probes into the nucleus for labeling the nucleus during the experimental procedure. There is a chip of coded probes designed based on photocleavable connectors (photocleavable linker), which usually requires light irradiation of the chip with corresponding wavelength bands to release coded probes, and this form of light generally covers the whole chip, resulting in release of probes under and outside the tissue, and thus, the phenomenon that probes outside the tissue enter the nucleus in the tissue area and the positioning accuracy is lowered may occur. The probe releasing mode enables probes at all positions on the chip to fall off at random, so that the probes randomly enter the cell nucleus and become noise in the subsequent reverse transcription and library building processes, the uniqueness of the cell nucleus in-situ probes is affected, and the positioning accuracy of the cell nucleus is affected. Disclosure of Invention The embodiment of the application mainly aims to provide a method and a system for releasing a space histology chip probe based on image guidance so as to improve the accuracy of probe release. To achieve the above object, an aspect of an embodiment of the present application provides an image-guided spatial histology chip probe release method, which includes the steps of: shooting an original image of a tissue slice by using a camera, wherein the tissue slice is attached to the surface of a probe coding chip; preprocessing the original image so as to locate and obtain a tissue region in the original image; Converting the tissue region as a tissue mask from camera coordinates to digital micromirror device coordinates; generating a control signal of the digital micro-mirror device according to the coordinates of the digital micro-mirror device; and responding to the control signal, and utilizing the digital micro-mirror device to project light generated by a light source to the tissue region so as to release the corresponding probes in the tissue region range in the probe coding chip. In some embodiments, the preprocessing the original image to locate the tissue region in the original image includes the steps of: identifying tissue contours in the original image using threshold segmentation, a deep learning model, or edge detection and morphological processing; Binarizing the original image according to the tissue outline to distinguish and obtain a tissue area and a non-tissue area in the original image; And correcting the edge of the tissue region by using morphological opening and closing operation so as to enable the mask corresponding to the tissue region to be continuous and smooth in illumination boundary. In some embodiments, the generating the control signal of the digital micromirror device according to the coordinates of the digital micromirror device includes the steps of: distributing switching instructions of the micromirror array according to different assignment of coordinates of the digital micromirror device; Outputting the switch instruction to a micromirror state matrix; Converting the micromirror state matrix into a binary image frame, writing the binary image frame into a display buffer of a controller of the digital micromirror device, and starting a single-frame static projection mode; And generating the control signal synchronously triggered by the light source and the digital micro-mirror device. In some embodiments, the method further comprises the steps of: Selecting the light source matched with the characteristics of the light fracture connection sub, wherein the wavelength of the light source comprises an ultraviolet band, a visible light band or a near infrared band; and a shading component or a scattering suppression module is arranged in the light path system of the light source so as to reduce the influence of stray light or reflected light. In some embodiments, the projecting light generated by a light source onto the tissue region using the digital micromirror device in response to the control signal to release a corresponding probe within the tissue region in the probe encoding chip comprises the steps of: Switching an inclined state by using a micromirror array on the digital micromirror device according to the control signal in response to the control signal to realize spatial modulation of an optical signal, wherein the inclined state comprises on and off; When the tilting state of the micromirror array is on, controlling incident light to reflect to a projection light path, so that the m