Cell manipulation observation device and cell manipulation observation method

Abstract

The invention discloses a cell manipulation observation device and a cell manipulation observation method.According to the cell manipulation observation device, an initial light beam is emitted through a light source module, the initial light beam is split through a light splitting module to form a first light beam and a second light beam, and at the moment, the light intensity of the first light beam and the light intensity of the second light beam are distributed through the light splitting module; the upper polar plate or the lower polar plate is used for irradiating the micro-fluidic chip; after the second light beam irradiates the lower polar plate, dielectrophoretic force is generated through the photoconductive layer to drive the cells to move, so that the cells can be effectively moved, and the light intensity can be reduced to prevent the cells from being damaged. The first light beam irradiates the upper polar plate and penetrates through the upper polar plate to irradiate specific cells in the micro-channel, so that the specific cells or secretions of the specific cells can be effectively and conveniently identified and observed. By arranging the first light path module and the second light path module, the paths of the first light beam and the second light beam can be changed, so that corresponding adjustment can be performed based on specific requirements, and control and observation of different types of micro-fluidic chips can be met.

Description

Technical Field

[0001] The present invention relates to the field of microfluidics technology, and in particular to a cell manipulation observation device and a cell control method. Background Art

[0002] Currently, transistor-based photoelectric tweezers technology has been used to manipulate (e.g., select or move) micro-objects such as cells, bacteria, and microspheres. The typical structure of this type of photoelectric tweezers device is to set up a microfluidic channel between two upper and lower electrodes, where the upper electrode is usually a glass plate coated with indium tin oxide (ITO), and the lower electrode is a metal electrode with a phototransistor array on it to replace the ordinary photoelectric layer. When patterned light is irradiated on a specific area on the phototransistor array, the activated transistors allow current to pass, thereby forming a non-uniform electric field in the microfluidic channel, generating a dielectrophoresis (DEP) force that can manipulate cells.

[0003] In existing technologies, the magnitude of the DEP force is limited by factors such as the cell's volume and the medium in which it resides. For a given analyte, increasing the illumination intensity and / or illuminated area of ​​the phototransistor can generate a larger photogenerated current, thereby creating a more pronounced non-uniform electric field and, consequently, a stronger DEP manipulation force. However, excessive illumination intensity can damage cells, and precise screening requires observation of microscopic objects. Therefore, a solution is needed that both protects cells and enables effective observation. Summary of the Invention

[0004] Based on this, it is necessary to provide a cell manipulation and observation device to address the existing problems, which is used to manipulate and observe cells on a microfluidic chip, including:

[0005] The microfluidic chip comprises an upper electrode plate and a lower electrode plate, wherein the upper surface of the lower electrode plate is provided with a photoconductive layer;

[0006] An illumination system includes a light source module, a spectroscopic module, a first optical path module, and a second optical path module. The light source module emits an initial light beam to the spectroscopic module. The spectroscopic module splits the initial light beam into a first light beam and a second light beam. The first light beam irradiates the upper electrode plate via the first optical path module, and the second light beam irradiates the lower electrode plate via the second optical path module. The second light beam irradiates the photoconductive layer to generate a dielectrophoretic force to drive cell movement.

[0007] An observation system is used to observe cells on the microfluidic chip.

[0008] Preferably, the light splitting module includes a switching component and a light splitting element, the light splitting element is used to split the initial light beam with the light intensity adjusted according to a preset ratio and form the first light beam and the second light beam, and the light splitting element is detachably mounted on the switching component.

[0009] Preferably, there are multiple light splitting elements, and the multiple light splitting elements are arranged on the switching component, and the switching component can switch the light splitting elements with different splitting ratios.

[0010] Preferably, the first optical path module includes a light intensity adjustment structure for adjusting the light intensity of the first light beam, a reflection structure for adjusting the light path direction of the first light beam, and a beam combining structure for combining the first light beams. The first light beam passes through the light intensity adjustment module, the reflection structure and the beam combining structure in sequence and then irradiates the upper electrode plate.

[0011] Preferably, the observation system includes an objective lens, a tube lens and a photographing device, the objective lens is located between the beam combining structure and the upper plate, the first light beam passes through the objective lens, and the tube lens and the photographing device are used to observe cells.

[0012] Preferably, the observation system further includes a detection module for detecting specific substances, and a control module for analyzing detection information and manipulating the shooting device, wherein the control module can control the shooting device to track and shoot the specific substances detected by the detection module.

[0013] Preferably, the first optical path module can change the optical path of the first light beam, the second optical path module can change the optical path of the second light beam, and the control module can control the first optical path module to irradiate the first light beam at a specific material distribution.

[0014] The present application also provides a cell observation control method for operating and observing cells on a microfluidic chip, comprising the following steps:

[0015] S01, providing a light source module and a light splitting module, wherein the light source module emits an initial light beam to the light splitting module, and the light splitting module splits the initial light beam into a first light beam and a second light beam according to a preset ratio;

[0016] S02, providing a first optical path module and a second optical path module, wherein the first light beam is transmitted through the first optical path module to irradiate the upper electrode plate of the microfluidic chip, and the second light beam is transmitted through the second optical path module to irradiate the lower electrode plate of the microfluidic chip, wherein the upper surface of the lower electrode plate is provided with a photoconductive layer;

[0017] S04, providing an observation system, wherein the observation system is positioned in a field of view where the cells are located to obtain cell images.

[0018] Preferably, the observation system includes an objective lens, a tube lens and a shooting device, and the objective lens is arranged between the upper electrode plate of the microfluidic chip and the first optical path module.

[0019] Compared with the prior art, the technical solution disclosed in the present invention has the following beneficial effects:

[0020] 1. The initial light beam emitted by the light source module is split into a first light beam and a second light beam through a spectrometer module. At this time, the light intensities of the first light beam and the second light beam are distributed through the spectrometer module to irradiate the upper plate or the lower plate of the microfluidic chip; the second light beam irradiates the lower plate, and generates a dielectric electrophoresis force through the photoconductive layer to drive the cells to move, which can not only achieve effective movement of the cells, but also reduce the light intensity to prevent damage to the cells; 2. The first light beam irradiates the upper plate, and passes through the upper plate to irradiate specific cells in the microchannel, which can effectively facilitate the identification and observation of specific cells or secretions of specific cells; 3. By setting the first optical path module and the second optical path module, the paths of the first light beam and the second light beam can be changed, so that corresponding adjustments can be made based on specific needs, which can meet the control and observation of different types of microfluidic chips. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] A more complete understanding of the exemplary embodiments of the present invention can be obtained by referring to the following drawings. The drawings are intended to provide a further understanding of the embodiments of the present application and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the present invention and do not constitute a limitation of the present invention. In the drawings, the same reference numerals generally represent the same components or steps.

[0022] Figure 1 This is a schematic structural diagram of a cell manipulation and observation device provided according to an exemplary embodiment of the present application;

[0023] Figure 2 A schematic structural diagram of a light splitting module provided according to an exemplary embodiment of the present application;

[0024] Figure 3 A schematic structural diagram of a light splitting element provided by an exemplary embodiment of the present application is shown.

[0025] Reference numerals

[0026] 101-light source; 102-focusing lens; 103-field stop; 104-aperture stop;

[0027] 105-spectroscopy module; 106-first reflector; 107-first condenser;

[0028] 108-lens; 109-second reflecting mirror; 110-second condensing mirror; 111-beam combining mirror;

[0029] 112-objective lens; 113-microfluidic chip; 114-observation system; 201-spectrometer;

[0030] 202 - switching component; 301 - first right-angle prism; 302 - second right-angle prism. DETAILED DESCRIPTION

[0031] Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present disclosure are shown in the accompanying drawings, it should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments described herein. Rather, these embodiments are provided to enable a more thorough understanding of the present disclosure and to fully convey the scope of the present disclosure to those skilled in the art.

[0032] In the description of the present invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings and are intended solely to facilitate and simplify the description of the present invention. They are not intended to indicate or imply that the devices or components referred to must have, be constructed, or operate in a specific orientation, and therefore should not be construed as limitations on the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0033] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed, detachable, or integral connections; mechanical or electrical connections; direct or indirect connections through an intermediate medium; and internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances.

[0034] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0035] Example 1

[0036] See also Figure 1, an embodiment of the present application provides a cell manipulation and observation device for manipulating and observing cells on a microfluidic chip 113, comprising a microfluidic chip 113, an illumination system, and an observation system; wherein the microfluidic chip 113 comprises an upper electrode plate (not shown) and a lower electrode plate (not shown), and a photoconductive layer is provided on the upper surface of the lower electrode plate, and the photoconductive layer can be a single layer, a multilayer, or a patterned structure; the illumination system comprises a light source module, a spectroscopic module 105, a first optical path module, and a second optical path module, the light source module emits an initial light beam to the spectroscopic module 105, the spectroscopic module 105 splits the initial light beam into a first light beam and a second light beam, the first light beam irradiates the upper electrode plate through the first optical path module, the second light beam irradiates the lower electrode plate through the second optical path module, and the second light beam irradiates the photoconductive layer to generate a dielectrophoretic force to drive the cells to move; and an observation system for observing cells on the microfluidic chip 113.

[0037] The embodiments of the present application have the following beneficial effects: 1. The initial light beam emitted by the light source module is split into a first light beam and a second light beam through the spectrometer module 105. At this time, the light intensities of the first light beam and the second light beam are distributed through the spectrometer module 105. The second light beam irradiates the lower electrode plate and generates a dielectric electrophoresis force through the photoconductive layer to drive the cells to move, which can not only achieve effective movement of the cells, but also reduce the light intensity to prevent damage to the cells; 2. The first light beam irradiates the upper electrode plate, and passes through the upper electrode plate to irradiate specific cells in the microchannel, which can effectively facilitate the identification and observation of specific cells or secretions of specific cells; 3. By setting the first optical path module and the second optical path module, the paths of the first light beam and the second light beam can be changed, so that corresponding adjustments can be made based on specific needs, which can meet the control and observation of different types of microfluidic chips 113.

[0038] Specifically, the photoconductive layer can be made of one or more of: hydrogenated amorphous silicon (a-Si:H), titanium phthalocyanine (TiOPC), CulnSe2, cadmium sulfide (CdS), perovskite material, quantum dot material, etc. It can be a photoconductive material of a single material or a photoconductive material of a composite material. As long as the dielectrophoretic force can be generated through the photoconductive layer, there is no sole limitation on the specific material and ratio of the photoconductive layer.

[0039] In addition, the photoconductive layer can form PNP (phototransistor), NPN (phototransistor) or PIN (photodiode) structures on the substrate of the lower electrode plate. No unique limitation is made here on the specific structure formed by the photoconductive layer on the substrate of the lower electrode plate.

[0040] Specifically, the light source module is a light source 101 with adjustable light intensity. The intensity of the initial light beam emitted by light source 101 can be adjusted according to the operating environment. The light source module includes light source 101, focusing lens 102, field stop 103, and aperture stop 104. The initial light beam emitted by light source 101 passes through focusing lens 102, then through field stop 103 and aperture stop 104 in sequence, and reaches beam splitting module 105. After passing through beam splitting module 105, the light beam is split into two parts, forming a first light beam and a second light beam.

[0041] Preferably, in one embodiment, Figure 3 As shown, the beam splitting module 105 includes a switching assembly 202 and a beam splitter 201. The beam splitter 201 is used to split the initial light beam by adjusting the intensity according to a preset ratio, forming a first beam and a second beam. The beam splitter 201 is detachably mounted on the switching assembly 202. The intensity distribution of the first and second beams can be adjusted automatically according to different usage requirements. The beam splitter 201 can be a beam splitter prism. The beam splitter prism is composed of two right-angle prisms 301, which are coated at the center to achieve energy beam splitting. Figure 3 This is a schematic diagram of a beam splitter prism, which includes two right-angle prisms 301. After a light beam passes through the prism, it is split into a first light beam and a second light beam of two energies. The energy ratio of the first light beam to the second light beam can be controlled by the coating on the two right-angle prisms 301.

[0042] Specifically, according to different requirements for adjusting the light intensity, the light splitting element 201 can be set to be replaceable, and a plurality of light splitting elements 201 are set on the switching component 202, and the switching component 202 can switch the light splitting elements 201 with different splitting ratios. Figure 2 As shown, the switching assembly 202 can be configured as a guide rail, with multiple beam splitters 201 arranged side by side on the guide rail. A specific individual beam splitter 201 can be positioned along the optical path based on actual needs. Of course, the switching assembly 202 can also be configured as a detachable structure, with the beam splitter 201 removably installed within the switching assembly 202. The beam splitter module 105 can also be configured as another device with manually adjustable light intensity, such as an acousto-optic modulator (AOM) or a polarization beam splitter. As long as the initial light beam can be split into the first and second light beams as required, and the light intensity distribution can be adjusted as required, the specific structure of the beam splitter module 105 is not limited here.

[0043] Preferably, in one embodiment, Figure 1As shown, the first optical path module includes a light intensity adjustment structure for adjusting the intensity of the first light beam, a reflective structure for adjusting the optical path direction of the first light beam, and a beam combining structure for combining the first light beam. The first light beam passes through the light intensity adjustment module, the reflective structure, and the beam combining structure in sequence before irradiating the upper electrode plate. When it is necessary to observe cells, the first and second light beams may still damage the cells after passing through the spectrometer module 105. To prevent the first light beam from damaging the cells due to excessive light intensity, the light intensity adjustment structure can be used to adjust the light intensity of the first light beam to a lower level, and then the light intensity of the first light beam can be enhanced according to actual observation needs. This can not only protect the cells from damage, but also effectively ensure the observation effect.

[0044] Preferably, in one embodiment, Figure 1 As shown, the observation system includes an objective lens 112, a tube lens, and a shooting device. The objective lens 112 is located between the beam combining structure and the upper plate. The first light beam passes through the objective lens 112. The tube lens and the shooting device are used to observe cells. After passing through the beam splitter 201, the first light beam passes through the lens 108, the second reflector 109, and the second condenser 110. It is combined with the imaging light beam of the objective lens 112 by the beam combiner 111 and enters the objective lens 112. After passing through the objective lens 112, the first light beam illuminates the upper surface of the microfluidic chip 113. The first light beam reflected or transmitted by the biochip to be photographed passes through the beam combiner 111 to the tube lens and shooting device of the microscope system. The tube lens, objective lens 112, and camera constitute an observation system that can observe the cell state.

[0045] Preferably, in one embodiment, the observation system further includes a detection module for detecting specific substances, and a control module for analyzing detection information and manipulating the shooting device, and the control module can control the shooting device to track and shoot the specific substances detected by the detection module. Specifically, specific cells or secretions of specific cells can be fluorescently labeled, and the observation system includes a fluorescence module, which can capture fluorescence, and the fluorescence intensity is analyzed by the control module in the observation system. During cell movement, it can be tracked and recorded by the shooting device and the fluorescence module. The control module can realize the simultaneous manipulation of cells and real-time imaging. In addition, based on the preset manipulation route, the control module can predict the actual movement trajectory of the cells and predict the optimal observation area through the control module.

[0046] Preferably, in one embodiment, Figure 1As shown, the first optical path module can change the optical path of the first light beam, and the second optical path module can change the optical path of the second light beam. The control module can control the first optical path module to illuminate the first light beam at a specific material distribution location. The operator can preset the microfluidic chip 113 to a predetermined detection position based on actual site requirements, and then use the first optical path module to change the optical path of the first light beam so that it illuminates the upper electrode plate. Similarly, the second optical path module can change the optical path of the second light beam so that it illuminates the lower electrode plate.

[0047] Specifically, the first optical path module includes a lens 108, a second reflector 109, and a condenser 110. Through a beam combiner 111 and an objective lens 112, the optical path is changed and the intensity of the light is adjusted. Similarly, the second optical path module includes a first reflector 106 and a first condenser 107 for changing the optical path.

[0048] Example 2

[0049] The present embodiment further provides a cell manipulation and observation method for manipulating and observing cells on the microfluidic chip 113, comprising the following steps:

[0050] S01, first place the microfluidic chip 113 at a preset position, then provide a light source module and a spectrometer module 105, the light source module emits an initial light beam to the spectrometer module 105, and the spectrometer module 105 splits the initial light beam into a first light beam and a second light beam according to a preset ratio;

[0051] S02, providing a first optical path module and a second optical path module, wherein the first light beam passes through the first optical path module to illuminate the upper electrode plate of the microfluidic chip 113, and the second light beam passes through the second optical path module to illuminate the lower electrode plate of the microfluidic chip 113, wherein the upper surface of the lower electrode plate is provided with a photoconductive layer;

[0052] S04, providing an observation system 114, positioning the observation system 114 at a field of view where the cells are located to obtain cell images.

[0053] Preferably, in one embodiment, the observation system 114 includes an objective lens 112 , a tube lens, and a photographing device, and the objective lens 112 is disposed between the upper plate of the microfluidic chip 113 and the first optical path module.

[0054] It can be understood that the same or similar parts of the above embodiments can be referenced to each other, and the contents not described in detail in some embodiments can refer to the same or similar contents in other embodiments.

[0055] It should be noted that, in the description of the present invention, the terms "first", "second", etc. are used for descriptive purposes only and should not be understood as indicating or implying relative importance. In addition, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is at least two.

[0056] Throughout this specification, reference to terms such as "one embodiment," "some embodiments," "examples," "specific examples," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0057] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A cell manipulation and observation device for manipulating and observing cells on a microfluidic chip, characterized in that: include: The microfluidic chip comprises an upper electrode plate and a lower electrode plate, wherein the upper surface of the lower electrode plate is provided with a photoconductive layer; An illumination system includes a light source module, a spectroscopic module, a first optical path module, and a second optical path module. The light source module emits an initial light beam to the spectroscopic module, and the spectroscopic module splits the initial light beam into a first light beam and a second light beam. The first light beam irradiates the upper electrode plate via the first optical path module, and the second light beam irradiates the lower electrode plate via the second optical path module. The second light beam irradiates the photoconductive layer to generate a dielectrophoretic force to drive cell movement. as well as An observation system is used to observe cells on the microfluidic chip.

2. The cell manipulation observation device according to claim 1, wherein: The light splitting module includes a switching component and a light splitting element. The light splitting element is used to split the initial light beam to adjust the light intensity according to a preset ratio and form the first light beam and the second light beam. The light splitting element can be replaceably installed on the switching component.

3. The cell manipulation observation device according to claim 2, wherein: There are multiple light splitting elements, and the multiple light splitting elements are arranged on the switching component, and the switching component can switch the light splitting elements with different light splitting ratios.

4. The cell manipulation observation device according to claim 1, wherein: The first optical path module includes a light intensity adjustment structure for adjusting the light intensity of the first light beam, a reflection structure for adjusting the light path direction of the first light beam, and a beam combining structure for combining the first light beams. The first light beam passes through the light intensity adjustment module, the reflection structure and the beam combining structure in sequence and then irradiates the upper electrode plate.

5. The cell manipulation observation device according to claim 1, wherein: The observation system includes an objective lens, a tube lens, and a shooting device. The objective lens is located between the beam combining structure and the upper plate. The first light beam passes through the objective lens. The tube lens and the shooting device are used to observe cells.

6. The cell manipulation observation device according to claim 5, wherein: The observation system further includes a detection module for detecting a specific substance, and a control module for analyzing detection information and manipulating the photographing device. The control module can control the photographing device to track and photograph the specific substance detected by the detection module.

7. The cell manipulation observation device according to claim 6, wherein: The first optical path module can change the optical path of the first light beam, the second optical path module can change the optical path of the second light beam, and the control module can control the first optical path module to irradiate the first light beam at a specific material distribution.

8. A cell manipulation observation method, characterized in that: Used to operate and observe cells on a microfluidic chip, including the following steps: S01, providing a light source module and a light splitting module, wherein the light source module emits an initial light beam to the light splitting module, and the light splitting module splits the initial light beam into a first light beam and a second light beam according to a preset ratio; S02, providing a first optical path module and a second optical path module, wherein the first light beam is transmitted through the first optical path module to irradiate the upper electrode plate of the microfluidic chip, and the second light beam is transmitted through the second optical path module to irradiate the lower electrode plate of the microfluidic chip, wherein the upper surface of the lower electrode plate is provided with a photoconductive layer; S04, providing an observation system, wherein the observation system is positioned in a field of view where the cells are located to obtain cell images.

9. The cell manipulation observation method according to claim 8, wherein: The observation system includes an objective lens, a tube lens and a shooting device. The objective lens is arranged between the upper electrode plate of the microfluidic chip and the first optical path module.

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