Living cell imaging light path system

Abstract

The invention belongs to the field of medical detection instruments, and discloses a living cell imaging light path system which comprises an LED straight lens cone, a fluorescent base, a filtering imaging lens cone, a camera reflection base, a camera mounting seat and an imaging camera. A light source is emitted from the LED straight lens cone to generate emitted light, the emitted light is reflected by the reflector, penetrates through the objective lens to be irradiated to a sample and is excited by cells to generate exciting light, the exciting light is reflected back, penetrates through the filter light imaging lens cone, is reflected by the camera reflection base and then is absorbed by the imaging camera through the camera mounting base, and therefore a cell image is read. The solid-state LED light source is applied, so that lightening at the exposure moment can be realized, the phototoxicity is reduced, the dynamic change of living cells can be accurately observed in real time, the observation efficiency is effectively improved, and errors and mistakes caused by manual observation are reduced. In addition, the living cell imaging light path system is small in structural size, convenient to install and capable of being matched with various living cell analyzers.

Description

Technical Field

[0001] The present invention belongs to the field of medical detection instruments, and more specifically, relates to a living cell imaging optical path system. Background Art

[0002] A live cell imager is a scientific research device that combines optical imaging, fluorescent labeling, and automation technology. It can observe the dynamic changes of living cells in real time and is widely used in fields such as cell biology, drug development, and clinical diagnosis. For example, by labeling specific cell structures (such as mitochondria and cell nuclei) with fluorescent proteins or dyes, dynamic signals can be captured, and real-time processes such as cell division, apoptosis, and migration (such as scratch experiments) can be tracked. This allows for the evaluation of gene transfection efficiency, drug toxicity, and efficacy, and supports high-throughput 384-well plate experiments.

[0003] Currently, most live-cell imaging instruments have certain flaws in their optical pathways. Fluorescence imaging relies on excitation light to activate fluorescent markers, but short wavelengths (such as ultraviolet light) and high-intensity light can cause phototoxicity, leading to decreased cell activity or even cell death. They also have only a single-color fluorescence channel, allowing only single-color fluorescence signals to be observed. Furthermore, multi-channel LED fluorescence can interfere with each other. Therefore, there is an urgent need to design a live-cell imaging optical system that can accurately observe the dynamic changes of living cells in real time. Summary of the Invention

[0004] In response to the above-mentioned defects or improvement needs of the existing technology, the present invention provides a living cell imaging optical path system, which aims to accurately observe the dynamic changes of living cells in real time, thereby solving the technical problems that short wavelength and high-intensity light lead to decreased cell activity or death, a single color channel cannot observe multiple fluorescence signals, and multi-channel LED fluorescence causes light source interference.

[0005] To achieve the above object, according to one aspect of the present invention, a living cell imaging optical system is provided, which comprises: an LED straight lens barrel, a fluorescence base, a filter imaging lens barrel, a camera reflective base, a camera mounting base and an imaging camera;

[0006] The LED straight lens barrel is used to emit different LED fluorescent light sources;

[0007] The fluorescent base is used to transmit light emitted by different LED fluorescent light sources and reflect it onto the cell sample to excite the cells to generate excitation light, and then reflect the excitation light to the filter imaging lens barrel;

[0008] The filter imaging lens barrel filters the excitation light;

[0009] The camera reflective base reflects the filtered excitation light to the camera mounting base and the imaging camera;

[0010] The camera mounting base and the imaging camera are used to finally receive the excitation light and read the cell image.

[0011] Preferably, the LED straight lens barrel includes a plano-convex lens 1 , an LED lamp bead 2 and a fixed lens barrel 3 , the plano-convex lens 1 is fixed in front of the fixed lens barrel 3 , and the LED lamp bead 2 is fixed behind the fixed lens barrel 3 .

[0012] Preferably, the LED lamp bead 2 includes three wavelengths of LED light sources, namely 570nm yellow, 470nm blue, and 385nm purple, which are used to excite dyes of different colors in cells respectively.

[0013] Preferably, the light source control of the LED lamp bead 2 is controlled by the Hi7012 chip in the lower computer circuit board, thereby controlling and adjusting the brightness of the LED light source.

[0014] Preferably, the fluorescent base includes a reflector 4, a mounting base 5, a dichroic mirror 1 6, a dichroic mirror 2 7, and a dichroic mirror 3 8; the dichroic mirror 1 6, the dichroic mirror 2 7, and the dichroic mirror 3 8 are respectively inserted into the gaps of the mounting base 5, and the reflector 4 is fixed on the lower side of the mounting base 5.

[0015] Preferably, the optical filtering imaging lens barrel comprises an optical filter 9, a tube lens and a mounting seat 10, and a lens barrel 11, and the optical filter 9, the tube lens and the mounting seat 10, and the lens barrel 11 are connected in sequence.

[0016] Preferably, the filter 9 is an achromatic doublet lens, which is used to correct chromatic aberration, suppress spherical aberration, and control light loss of the reflected cell excitation light.

[0017] Preferably, the camera reflective base includes a camera reflector 12 and a reflector mounting seat 13. The camera reflector 12 is mounted on the reflector mounting seat 13. The camera reflective base reflects the excitation light perpendicular to the direction of the camera through a 45° mirror, and converts it into light parallel to the direction of the camera, so that it is absorbed and read by the camera.

[0018] Preferably, the camera mount and the imaging camera include an imaging camera 14 and a camera mount 15 , the imaging camera 14 is mounted on the camera mount 15 , and the excitation light passes through the camera mount 15 and is absorbed by the imaging camera 14 , thereby reading the cell image.

[0019] In general, compared with the prior art, the above technical solution conceived by the present invention provides a living cell imaging optical system with the following beneficial effects:

[0020] 1. The present invention provides a living cell imaging optical path system comprising an LED straight lens barrel, a fluorescence base, a filter imaging lens barrel, a camera reflective base, a camera mounting base, and an imaging camera. The light source is emitted from the LED straight lens barrel to generate emission light, which is reflected by a reflector, passes through an objective lens, and strikes a sample. The cells are excited to generate excitation light. After the excitation light is reflected back, it passes through the filter imaging lens barrel, is reflected by the camera reflective base, and then passes through the camera mounting base to be absorbed by the imaging camera, thereby reading the cell image. The use of a solid-state LED light source can achieve instantaneous lighting during exposure, reduce phototoxicity, accurately observe the dynamic changes of living cells in real time, effectively improve observation efficiency, and reduce errors and mistakes caused by manual observation.

[0021] 2. The present invention provides a live-cell imaging optical system that uses three wavelengths of LED fluorescent lamps, yellow (570nm), blue (470nm), and violet (385nm), to excite different stained observation cells, thereby reducing impurities in the observation cells. The live-cell imaging optical system is also compact and easy to install. It can be adapted to various live-cell analyzers, such as single-point live-cell analyzers and scanning live-cell analyzers, to achieve clearer imaging results.

[0022] 3. The present invention provides a live cell imaging optical system that can be adapted to high-resolution, high-definition black-and-white cameras or color cameras. Combined with relevant image processing algorithms, it can intelligently and accurately reflect the morphology and dynamic changes of living cells, solving the problem of difficulty in achieving high resolution during live cell imaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a flowchart of the imaging process of a living cell imaging optical system of the present invention;

[0024] Figure 2 This is a schematic diagram of the overall structure of a living cell imaging optical system of the present invention;

[0025] Figure 3 This is a schematic diagram of the LED straight lens barrel structure in a living cell imaging optical path system of the present invention;

[0026] Figure 4 This is a schematic diagram of the structure of a fluorescent base in a living cell imaging optical system of the present invention;

[0027] Figure 5 This is a schematic diagram of the structure of a filter imaging lens barrel in a living cell imaging optical system of the present invention;

[0028] Figure 6 This is a schematic structural diagram of a camera reflective base in a living cell imaging optical system of the present invention;

[0029] Figure 7This is a schematic diagram of the structure of a camera mounting base and an imaging camera in a living cell imaging optical path system of the present invention;

[0030] Figure 8 Schematic diagram of the generation and propagation of 570nm yellow emission light in the present invention;

[0031] Figure 9 Schematic diagram of the generation and propagation of 470nm blue emission light in the present invention;

[0032] Figure 10 Schematic diagram of the generation and propagation of 385nm purple emission light in the present invention;

[0033] Figure 11 A schematic diagram of the generation and propagation of reflected light in the present invention;

[0034] Figure 12 This is the first practical application imaging effect of a living cell imaging optical system of the present invention;

[0035] Figure 13 This is the second imaging effect of the practical application of the living cell imaging optical system of the present invention.

[0036] Throughout the drawings, the same reference numerals are used to denote the same elements or structures, wherein:

[0037] 1- Plano-convex lens; 2- LED lamp bead; 3- Fixed lens barrel; 4- Reflector; 5- Mounting base; 6- Binochroic mirror 1; 7- Binochroic mirror 2; 8- Binochroic mirror 3; 9- Filter; 10- Tubular lens mount and tube lens; 11- Lens barrel; 12- Camera reflector; 13- Reflector mount; 14- Imaging camera; 15- Camera mount; 16-570nm yellow LED; 17-570nm yellow LED fixed lens barrel; 18-470nm blue LED; 19-470nm blue LED fixed lens barrel; 20-385nm violet LED; 21-385nm violet LED fixed lens barrel. DETAILED DESCRIPTION

[0038] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely for the purpose of explaining the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

[0039] like Figure 1 and Figure 2As shown, this embodiment provides a live cell imaging optical system, including an LED straight lens, a fluorescence base, a filter imaging lens, a camera reflective base, a camera mount, and an imaging camera. Light is emitted from the LED straight lens, generating emitted light. This light is reflected by a reflector, passes through an objective lens, and strikes the sample, where it is excited by the cells to produce excitation light. This excitation light is then reflected back through the filter imaging lens, reflected by the camera reflective base, and then passed through the camera mount before being absorbed by the imaging camera. This allows for accurate, real-time observation of the dynamic changes in living cells.

[0040] like Figure 3 As shown, the LED straight lens barrel includes a plano-convex lens 1, an LED lamp bead 2, and a fixed lens barrel 3. The plano-convex lens 1 is fixed in front of the fixed lens barrel 3, and the LED lamp bead 2 is fixed behind the fixed lens barrel 3. The LED lamp bead 2 includes three wavelengths of LED light sources: 570nm yellow, 470nm blue, and 385nm purple, which are used to excite different colored dyes in cells, thereby reducing impurities in the image observed through fluorescence. The light source of LED lamp bead 2 is controlled by the Hi7012 chip on the lower computer circuit board, achieving precise control of the LED light source brightness, which can be adjusted in real time according to actual conditions.

[0041] like Figure 4 As shown, the fluorescent base includes a reflector 4, a mounting base 5, a dichroic mirror 1 6, a dichroic mirror 2 7, and a dichroic mirror 3 8. These mirrors are inserted into the gaps in the mounting base 5, and the reflector 4 is fixed to the lower side of the mounting base 5. These three dichroic mirrors can only reflect or transmit light of the wavelength specified by their model in a specific direction and cannot transmit light in other directions. This allows them to accurately transmit any desired color of LED light, reducing interference and saving space.

[0042] like Figure 5 As shown, the filter imaging lens barrel includes a filter 9, a tube lens and mounting base 10, and a lens barrel 11, which are sequentially connected. Filter 9 is an achromatic doublet lens that corrects chromatic aberration, suppresses spherical aberration, and controls light loss in the reflected cell excitation light, significantly improving imaging resolution and signal detection sensitivity, resulting in clearer signals received by the camera.

[0043] like Figure 6As shown, the camera reflector base includes a camera reflector 12 and a reflector mount 13, with the camera reflector 12 mounted on the reflector mount 13. The camera reflector base reflects excitation light perpendicular to the camera's direction through a 45-degree mirror, converting it into light parallel to the camera's direction, which is then absorbed and read by the camera. The camera reflector base connects to the filter imaging lens barrel for optical signal transmission and directly connects the camera mount to the imaging camera, preventing optical signal attenuation and loss due to an overly complex optical signal path.

[0044] like Figure 7 As shown, the camera mounting seat and the imaging camera include an imaging camera 14 and a camera mounting seat 15 , and the imaging camera 14 is mounted on the camera mounting seat 15 .

[0045] like Figure 8 As shown, it is a schematic diagram of the generation and propagation of 570nm yellow emission light. The LED straight lens barrel is connected to the mounting base 5. The 570nm yellow LED 16 emits yellow emission light. The light emitted by the LED is converged into parallel light through the 570nm yellow LED fixed lens barrel 17 and the plano-convex lens in the lens barrel. The dichroic mirror 1 6 can pass the 570nm yellow emission light. The dichroic mirror 2 7 can pass the 570nm yellow emission light. The dichroic mirror 3 8 can reflect the 570nm yellow emission light to the reflector 4. The reflector 4 then reflects the 570nm yellow emission light to the cells above, thereby irradiating the cells to generate excitation light.

[0046] like Figure 9 As shown, it is a schematic diagram of the generation and propagation of 470nm blue emission light. The 470nm blue LED 18 emits blue emission light, which is converged into parallel light by the 470nm blue LED fixing lens tube 19 and the plano-convex lens in the lens tube. The dichroic mirror 1 6 can reflect the 470nm blue emission light to the dichroic mirror 2 7. The dichroic mirror 2 7 can pass the 470nm blue emission light. The dichroic mirror 3 8 can reflect the 470nm blue emission light to the reflector 4. The reflector 4 then reflects the 470nm blue emission light to the cells above, thereby irradiating the cells to generate excitation light.

[0047] like Figure 10 As shown, it is a schematic diagram of the generation and propagation of 385nm purple emission light. The 385nm purple LED 20 emits purple emission light, which is converged into parallel light by the 385nm purple LED fixing lens barrel 21 and the plano-convex lens in the lens barrel. The dichroic mirror 2 7 can reflect the 385nm purple emission light, and the dichroic mirror 3 8 can reflect the 385nm purple emission light to the reflector 4. The reflector 4 then reflects the 385nm purple emission light to the cells above, thereby irradiating the cells to generate excitation light.

[0048] like Figure 11As shown in the figure, it is a schematic diagram of the generation and propagation of reflected light. After the excitation light is reflected back, it is reflected by the reflector 4, and then passes through the dichroic mirror 8. The excitation light passes through the filter imaging lens barrel, is reflected by the camera reflector 12, and then passes through the camera mounting base 15 and is absorbed by the imaging camera 14, thereby reading the cell image.

[0049] like Figure 12 、 13 As shown in FIG. 1 , the imaging effect of a live cell imaging optical path system in this embodiment applied to a live cell analyzer is shown. The cell image in the figure is clear, and the dynamic changes of live cells can be accurately observed in real time, with high observation efficiency, reducing errors and mistakes caused by manual observation.

[0050] It will be easily understood by those skilled in the art that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A living cell imaging optical system, characterized by: include: LED straight lens tube, fluorescent base, filter imaging lens tube, camera reflector base, camera mount and imaging camera; The LED straight lens barrel is used to emit different LED fluorescent light sources; The fluorescent base is used to transmit light emitted by different LED fluorescent light sources and reflect it onto the cell sample to excite the cells to generate excitation light, and then reflect the excitation light to the filter imaging lens barrel; The filter imaging lens barrel filters the excitation light; The camera reflective base reflects the filtered excitation light to the camera mounting base and the imaging camera; The camera mounting base and the imaging camera are used to finally receive the excitation light and read the cell image.

2. The living cell imaging optical system according to claim 1, wherein: The LED straight lens barrel comprises a plano-convex lens (1), an LED lamp bead (2) and a fixed lens barrel (3); the plano-convex lens (1) is fixed in front of the fixed lens barrel (3), and the LED lamp bead (2) is fixed in the rear of the fixed lens barrel (3).

3. The living cell imaging optical system according to claim 2, wherein: The LED lamp bead (2) comprises three wavelengths of LED light sources, namely 570nm yellow, 470nm blue, and 385nm purple, which are respectively used to excite dyes of different colors in cells.

4. The living cell imaging optical system according to claim 3, wherein: The light source control of the LED lamp bead (2) is performed through the Hi7012 chip in the lower computer circuit board, thereby controlling and adjusting the brightness of the LED light source.

5. The living cell imaging optical system according to claim 1, wherein: The fluorescent base comprises a reflector (4), a mounting base (5), a first dichroic mirror (6), a second dichroic mirror (7), and a third dichroic mirror (8); the first dichroic mirror (6), the second dichroic mirror (7), and the third dichroic mirror (8) are respectively inserted into the gaps of the mounting base (5), and the reflector (4) is fixed on the lower side of the mounting base (5).

6. The living cell imaging optical system according to claim 1, wherein: The filter imaging lens barrel comprises a filter (9), a tube lens and a mounting seat (10), and a lens barrel (11), wherein the filter (9), the tube lens and the mounting seat (10), and the lens barrel (11) are connected in sequence.

7. The living cell imaging optical system according to claim 6, wherein: The filter (9) is an achromatic doublet lens, which is used to correct chromatic aberration, suppress spherical aberration, and control light loss of the reflected cell excitation light.

8. The living cell imaging optical system according to claim 1, wherein: The camera reflective base comprises a camera reflective mirror (12) and a reflective mirror mounting seat (13). The camera reflective mirror (12) is mounted on the reflective mirror mounting seat (13). The camera reflective base reflects excitation light perpendicular to the direction of the camera through a 45° mirror, converting the light into light parallel to the direction of the camera, so that the light is absorbed and read by the camera.

9. The living cell imaging optical system according to claim 1, wherein: The camera mounting seat and the imaging camera include an imaging camera (14) and a camera mounting seat (15), wherein the imaging camera (14) is mounted on the camera mounting seat (15), and the excitation light passes through the camera mounting seat (15) and is absorbed by the imaging camera (14), thereby reading a cell image.

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