Imaging well, stage, two-photon microscope, and imaging method

Abstract

An imaging well (100), comprising: an adhesive layer (110), and a liquid layer (120) covering the adhesive layer (110), the adhesive layer (110) and the liquid layer (120) forming a well body. The adhesive layer (110) is used for adhering a tissue specimen, the liquid layer (120) allows photons and liquids to pass through, and the well body accommodates an imaging liquid. The imaging well (100) allows specimen tissue to be adhered, and the operation process is simple and reduces tissue damage, thus significantly improving imaging stability and quality. In addition, the imaging well (100) can maintain the physiological state of tissues for a long time, and is suitable for long-time dynamic imaging research. Moreover, a stage comprising the imaging well (100), a two-photon microscope, and an imaging method are also provided.

Description

An imaging groove, stage, two-photon microscope and imaging method Technical Field

[0001] This application relates to the field of optical imaging technology, and in particular to an imaging slot, a stage, a two-photon microscope, and an imaging method. Background Technology

[0002] Two-photon microscopy is an advanced biomedical imaging technique that utilizes the simultaneous absorption of two low-energy photons to excite fluorescent molecules, thereby achieving high-resolution imaging of deep tissues. This technique has significant applications in studying the dynamic processes of living tissues, disease progression, and drug effects.

[0003] However, traditional two-photon imaging methods typically require fixing tissue samples onto specific glass slides, which is not only cumbersome but also prone to causing mechanical damage to the tissue, affecting imaging results. Furthermore, existing technologies struggle to maintain tissue stability and physiological state during prolonged imaging, leading to decreased image quality. Additionally, the confined spaces of some organs, such as cervical lymph nodes, make direct exposure for imaging difficult, necessitating devices to adhere the tissue and ensure sufficient fluid above it for optimal imaging.

[0004] Summary of the Invention

[0005] Therefore, it is necessary to provide an imaging tank, stage, two-photon microscope, and imaging method that can effectively improve the stability and quality of two-photon imaging while reducing tissue damage, addressing the current technical shortcomings in the stability and imaging quality of two-photon imaging.

[0006] To solve the above problems, this application adopts the following technical solution:

[0007] One objective of this application is to provide an imaging tank, comprising: an adhesive layer and a liquid layer covering the adhesive layer, the adhesive layer and the liquid layer forming a tank body, the adhesive layer being used to adhere tissue samples, the liquid layer allowing photons and liquid to pass through, and the tank body being able to contain imaging liquid.

[0008] In some embodiments, the adhesive layer is made of a biocompatible material, including polydimethylsiloxane.

[0009] In some embodiments, the liquid layer is made of a transparent optical material, including glass or a highly transparent plastic.

[0010] In some embodiments, the imaging liquid is water or PBS buffer.

[0011] A second objective of this application is to provide a stage on which the imaging groove is fixed. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples, the liquid layer allows photons and liquid to pass through, and the groove can contain the liquid required for imaging.

[0012] A third objective of this application is to provide a two-photon microscope, including a stage on which the imaging groove is fixed. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples, and the liquid layer allows photons and liquid to pass through. The groove can contain the liquid required for imaging.

[0013] Fourthly, this application also provides an imaging method for the aforementioned two-photon microscope, comprising the following steps:

[0014] The tissue sample is adhered to the adhesive layer;

[0015] Imaging liquid is added through the liquid layer;

[0016] The imaging slot is fixed on the stage;

[0017] Perform two-photon imaging.

[0018] The present application adopts the above technical solution, and its beneficial effects are as follows:

[0019] The imaging tank, stage, two-photon microscope, and imaging method provided in this application include: an adhesive layer and a liquid layer covering the adhesive layer, the adhesive layer and the liquid layer forming a tank, the adhesive layer for adhering tissue samples, the liquid layer allowing photons and liquid to pass through, and the tank containing imaging liquid. The imaging tank provided in this application can adhere to sample tissues, has a simple operation procedure, reduces tissue damage, and significantly improves the stability and quality of imaging. Furthermore, this imaging tank can maintain the physiological state of tissues for extended periods, making it suitable for long-term dynamic imaging studies. Attached Figure Description

[0020] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the description of the embodiments of this application or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 is a schematic diagram of the imaging slot provided in Embodiment 1 of this application.

[0022] Figure 2 is a flowchart of the imaging method of the two-photon microscope provided in Embodiment 4 of this application. Detailed Implementation

[0023] The embodiments of this application are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain this application, and should not be construed as limiting this application.

[0024] In the description of this application, it should be understood that the terms "upper", "lower", "horizontal", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application.

[0025] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this application, "multiple" means two or more, unless otherwise explicitly specified.

[0026] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments.

[0027] Example 1

[0028] Please refer to Figure 1, which is a schematic diagram of the structure of the imaging tank 100 provided in the embodiment of this application. It includes an adhesive layer 110 and a liquid layer 120 covering the adhesive layer 110. The adhesive layer 110 and the liquid layer 120 form a tank. The adhesive layer 110 is used to adhere tissue samples, the liquid layer 120 allows photons and liquid to pass through, and the tank can contain imaging liquid.

[0029] In this embodiment, the adhesion layer 110 is made of a biocompatible material, which can firmly adhere to the tissue sample while maintaining the physiological state of the tissue, reducing movement during the imaging process.

[0030] Furthermore, the biocompatible material includes polydimethylsiloxane.

[0031] In this embodiment, the liquid layer 120 is made of a transparent material with good optical properties to ensure the optical quality of two-photon imaging.

[0032] Furthermore, the transparent optical material includes glass or a highly transparent plastic.

[0033] In this embodiment, the imaging liquid is water or PBS buffer.

[0034] The imaging chamber provided in this application can adhere to sample tissues, has a simple operation procedure, reduces tissue damage, and significantly improves the stability and quality of imaging. Furthermore, this imaging chamber can maintain the physiological state of tissues for extended periods, making it suitable for long-term dynamic imaging studies.

[0035] Example 2

[0036] This application also provides a stage on which the imaging groove is fixed. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples, the liquid layer allows photons and liquid to pass through, and the groove can contain the liquid required for imaging.

[0037] In this embodiment, the imaging groove is fixed to the stage using magnetic or mechanical means to ensure stable fixation of the imaging groove on the microscope stage.

[0038] The stage provided in this application has the aforementioned imaging groove fixed in place, which can adhere to sample tissues. The operation is simple and reduces tissue damage, significantly improving imaging stability and quality. Furthermore, this imaging groove can maintain the physiological state of tissues for extended periods, making it suitable for long-term dynamic imaging studies.

[0039] Example 3

[0040] This application also provides a two-photon microscope, including a stage on which the imaging groove is fixed. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples, the liquid layer allows photons and liquid to pass through, and the groove can contain the liquid required for imaging.

[0041] The two-photon microscope provided in this application has an imaging groove fixed on the stage, which can adhere to sample tissues. This simplifies the operation process, reduces tissue damage, and significantly improves imaging stability and quality. Furthermore, the imaging groove can maintain the physiological state of tissues for extended periods, making it suitable for long-term dynamic imaging studies.

[0042] Example 4

[0043] Please refer to Figure 2, which provides an imaging method for two-photon microscopy, comprising the following steps:

[0044] Step S110: Adhere the tissue sample to the adhesive layer.

[0045] It should be noted that: the adhesive layer 110 and the liquid layer 120 covering the adhesive layer 110 provided in this embodiment form a tank. The adhesive layer 110 can be used to adhere tissue samples, the liquid layer 120 can allow photons and liquid to pass through, and the tank can contain imaging liquid.

[0046] Step S120: Add imaging liquid through the liquid layer.

[0047] Step S130: Fix the imaging slot to the stage.

[0048] In this embodiment, the imaging groove is fixed to the stage using magnetic or mechanical means to ensure stable fixation of the imaging groove on the microscope stage.

[0049] Step S140: Perform two-photon imaging.

[0050] A stage is provided, on which the imaging groove is fixed. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples, the liquid layer allows photons and liquid to pass through, and the groove can hold the liquid required for imaging.

[0051] The imaging method for two-photon microscopy provided in this application involves adhering a tissue sample to the adhesion layer, adding an imaging liquid through the liquid layer, fixing the imaging groove on the stage, and then performing two-photon imaging. This method is simple to operate, reduces tissue damage, and significantly improves imaging stability and quality. Furthermore, the imaging groove can maintain the physiological state of the tissue for extended periods, making it suitable for long-term dynamic imaging studies.

[0052] The above are merely preferred embodiments of this application, and only specifically describe the technical principles of this application. These descriptions are only for explaining the principles of this application and should not be construed as limiting the scope of protection of this application in any way. Based on this explanation, any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application, as well as other specific embodiments of this application that can be conceived by those skilled in the art without creative effort, should be included within the scope of protection of this application.

Claims

1. An imaging groove, characterized in that, include: An adhesive layer and a liquid layer covering the adhesive layer, the adhesive layer and the liquid layer forming a tank, the adhesive layer for adhering tissue samples, the liquid layer allowing photons and imaging fluid to pass through, and the tank containing imaging fluid.

2. The imaging groove as described in claim 1, characterized in that, The adhesive layer is made of a biocompatible material, including polydimethylsiloxane.

3. The imaging groove as described in claim 1, characterized in that, The liquid layer is made of a transparent optical material, including glass or a highly transparent plastic.

4. The imaging groove as described in claim 1, characterized in that, The imaging liquid is water or PBS buffer.

5. A stage, characterized in that, The imaging groove is fixed on the stage. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples. The liquid layer allows photons and liquid to pass through. The groove can hold the liquid required for imaging.

6. A two-photon microscope, characterized in that, The device includes a stage on which the imaging groove is fixed. The imaging groove includes an adhesive layer and a liquid layer covering the adhesive layer. The adhesive layer and the liquid layer form a groove. The adhesive layer is used to adhere tissue samples, the liquid layer allows photons and liquid to pass through, and the groove can hold the liquid required for imaging.

7. An imaging method for a two-photon microscope as described in claim 6, characterized in that, Includes the following steps: The tissue sample is adhered to the adhesive layer; Imaging liquid is added through the liquid layer; The imaging slot is fixed on the stage; Perform two-photon imaging.

Images