CN213022779U - Three-dimensional cell visualization device - Google Patents
Three-dimensional cell visualization device Download PDFInfo
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- CN213022779U CN213022779U CN202021553865.8U CN202021553865U CN213022779U CN 213022779 U CN213022779 U CN 213022779U CN 202021553865 U CN202021553865 U CN 202021553865U CN 213022779 U CN213022779 U CN 213022779U
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Abstract
The utility model discloses a three-dimensional stereoscopic visualization device of cell, including light source, spectroscope, light filter, microscope, charge-coupled device (CCD), piezoelectric scanning controller, treater. The cell sample stimulated emission fluorescence reaches the CCD through the light splitting system, a sample image is recorded by the CCD, then image data is transmitted to the processor, the piezoelectric scanning controller enables a microscope focal plane to step along an optical axis to image three-dimensional sample section by section, and the image is subjected to deconvolution recovery processing and stereoscopic visualization in the processor. The utility model discloses can improve image resolution to can gather living body cell three-dimensional image in succession, help studying the interior dynamic process of thin.
Description
Technical Field
The utility model belongs to the technical field of the optical microscopy imaging, concretely relates to three-dimensional stereoscopic device of cell.
Background
Modern life science research is mostly carried out on a cellular level, and the cells are taken as research objects, long-term dynamic observation and quantitative analysis are carried out on the cells, and the research on signal transduction and regulation pathways of single cells is carried out.
The traditional two-dimensional manual observation quantification has the following defects: the two-dimensional imaging precision is low, the spatial information is lost, the structural relationship is fuzzy, the image has no three-dimensional sense, and the scientific research requirements cannot be met; the labor intensity is high, and the self-adaption phenomenon is easy to occur after the operator is tired, so that random errors are generated in the research result. Particularly, under a microscope field of view, it is more difficult to manually perform a specific search for thousands of targets or repeatedly observe a plurality of specific objects for a long time.
The current body imaging devices are mainly used: the optical chromatography microscope, the deformable lens or the zoom lens, the light field microscopic imaging and the like are used for detecting the sample, the sample scanning time is long, the background fluorescence interference is large, and the resolution ratio is low.
SUMMERY OF THE UTILITY MODEL
Therefore, it is necessary to provide an intelligent device for three-dimensional stereo reconstruction of a two-dimensional image of a cell, aiming at the problems of inaccurate two-dimensional manual observation quantification, large workload, long imaging time, large interference, low image resolution and the like of the existing imaging device.
A three-dimensional cell visualization device comprises a spectroscope, an optical filter, a microscope, a charge coupled device, a piezoelectric scanning controller and a processor;
the processor is respectively connected with the charge coupled device and the piezoelectric scanning controller;
the piezoelectric scanning controller is connected with the spectroscope;
the spectroscope is used for decomposing exciting light and emitting light emitted by cells under the action of the exciting light, the optical filter is used for filtering the emitting light, and the charge-coupled device is used for collecting the emitting light signal, obtaining a fluorescence image and sending the fluorescence image to the processor; the piezoelectric scanning controller controls the focal plane of the microscope to step along the optical axis to obtain a multilayer slice image of the cell image; and the processor performs deconvolution processing and stereoscopic visualization processing according to the fluorescence image and the multilayer slice image to obtain a three-dimensional image of the cell.
Furthermore, the cell three-dimensional visualization device also comprises a light source used for emitting exciting light, and the light source is connected with the processor.
Preferably, the beam splitter in the cellular three-dimensional visualization device includes a dichroic beam splitter.
Preferably, the microscope in the cell three-dimensional visualization device comprises a wide-field fluorescence microscope.
Further, the microscope structure of the three-dimensional cell visualization device comprises an objective lens.
The utility model discloses simple structure, use portably, follow the two-dimensional optical section image of microscope optical axis collection biological sample under the weak fluorescence condition, then use deconvolution image recovery algorithm to resume because of receiving the original image that the outer light of focus influences and become fuzzy, improve image quality to use three-dimensional image deconvolution result as the basis, use the demonstration that stereotactic technique realized three-dimensional image data. Parallel optical slice imaging is adopted, errors possibly caused by point-to-point imaging are overcome, and the imaging speed is improved; the wide field inverted fluorescence microscope and the charge coupled device can improve the image resolution and conveniently display an object image in real time; the deconvolution restoration processing is carried out on the two-dimensional microscopic image optical section by adopting a non-physical mathematical method, so that the object image is 'confocal', the image resolution and the quality are greatly improved, a plurality of images can be obtained for photosensitive cells under low illumination and long-time irradiation, the phenomena of photobleaching and poisoning cells do not exist, the living cells can be dynamically observed, the study on cell dynamics is facilitated, the method can be applied to the three-dimensional reconstruction from the two-dimensional section image of the cells to the three-dimensional image in the biological field, and can also be applied to the three-dimensional reconstruction and display of human organs, soft tissues and pathological change parts in the medical field.
Drawings
FIG. 1 is a schematic view showing the basic structure of a three-dimensional cell visualization apparatus.
Description of reference numerals: spectroscope 1, slide glass and cell 2, objective 3, piezoelectric scanning controller 4, processor 5, light filter 6, CCD camera 7, exciting light source 8.
Detailed Description
In order to make the above objects, features and advantages of the present invention more comprehensible, embodiments of the present invention are described in detail below with reference to the accompanying drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, as those skilled in the art will be able to make similar modifications without departing from the spirit and scope of the present invention.
Modern life science research is mostly carried out on a cellular level, and the cells are taken as research objects, long-term dynamic observation and quantitative analysis are carried out on the cells, and the research on signal transduction and regulation pathways of single cells is carried out. The traditional two-dimensional manual observation quantification has the following defects: the two-dimensional imaging precision is low, the spatial information is lost, the structural relationship is fuzzy, the image has no three-dimensional sense, and the scientific research requirements cannot be met; the labor intensity is high, and the self-adaption phenomenon is easy to occur after the operator is tired, so that random errors are generated in the research result. Particularly, under a microscope field of view, it is more difficult to manually perform a specific search for thousands of targets or repeatedly observe a plurality of specific objects for a long time. The current body imaging devices are mainly used: the optical chromatography microscope, the deformable lens or the zoom lens, the light field microscopic imaging and the like are used for detecting the sample, the sample scanning time is long, the background fluorescence interference is large, and the resolution ratio is low.
In one embodiment, as shown in fig. 1, there is provided a cellular three-dimensional visualization apparatus, comprising a spectroscope and an optical filter, a microscope, a charge-coupled device, a piezoelectric scanning controller, a processor;
the processor is respectively connected with the charge coupled device and the piezoelectric scanning controller;
the piezoelectric scanning controller is connected with the spectroscope;
the spectroscope is used for decomposing exciting light and emitting light emitted by cells under the action of the exciting light, the optical filter is used for filtering the emitting light, and the charge-coupled device is used for collecting the emitting light signal, obtaining a fluorescence image and sending the fluorescence image to the processor; the piezoelectric scanning controller controls the focal plane of the microscope to step along the optical axis to obtain a multilayer slice image of the cell image; and the processor performs deconvolution processing and stereoscopic visualization processing according to the fluorescence image and the multilayer slice image to obtain a three-dimensional image of the cell.
In one embodiment, a three-dimensional visualization device for cells is provided, comprising a light source emitting excitation light; a spectroscope for splitting the emission light and the excitation light, which is positioned below the glass slide and the objective lens which are loaded with the cell sample and above the light source; the filter is used for filtering exciting light and is positioned below the spectroscope; a charge-coupled device (CCD) for collecting the excitation light signal, which is positioned behind the optical filter; the piezoelectric scanning controller is used for controlling the microscope focal plane to step along the optical axis and imaging the signal by slicing one by one and is positioned between the spectroscope and the processor; and the processor is used for controlling the light source and deconvoluting the image to realize stereoscopic visualization, and is respectively connected with the light source, the CCD and the piezoelectric scanning controller.
In one embodiment, the cell sample stimulated emission fluorescence reaches the CCD through the light splitting system, a three-dimensional fluorescence image of the sample located in a microscope focal plane is recorded by the CCD, then image data are transmitted to the processor, the piezoelectric scanning controller enables the microscope focal plane to step along an optical axis to image the three-dimensional sample in a slicing mode, and the image is subjected to deconvolution recovery processing and stereoscopic visualization in the processor.
In one embodiment, the microscope is a wide field fluorescence microscope, including an objective lens.
In one embodiment, the optical axis is defined as the Z-axis and the microscope focal plane is defined as the XY-plane; the piezoelectric scanning controller is used for Z-axis piezoelectric scanning control.
In one embodiment, when the device works, the image acquisition software is used for controlling the wavelength and the exposure time of the monochromatic excitation light, the excitation light irradiates the cell sample through the dichroic beam splitter, the fluorescence emitted by the sample under excitation reaches the CCD through the same dichroic beam splitter and the optical filter, the fluorescence image of the three-dimensional sample at the focal plane of the microscope is recorded by the CCD, and then the image data is sent to the processor. And controlling a piezoelectric Z-axis scanning controller to step the focal plane of the microscope along the Z axis so as to image the three-dimensional sample slice by slice. Finally, the acquired image is deconvoluted and restored and stereoscopically visualized in a processor.
In one embodiment, the microscope is an Olympus inverted microscope IX70, Japan, and the objective lens is UPIanaapo 100x/1.35NA Oil Iris from the Olympus APO l00x/1.65NA Oil HR series.
In one embodiment, the light source and the controller are Till Photonics GmBH of POLYCHROME IV series, the CCD is SVGA PCO of SensiCam company, Germany, the common PC, the piezoelectric Z-axis scanning controller is Physik instruments of E-662 LR series, Germany, and the image acquisition software is written by Python in combination with C + +.
In one embodiment, the wide-field microscope can be modified and installed based on the hardware description. After the installation, start collection control software control light source, Z axle scanning controller and obtain cell optical section, start the deconvolution function, and then carry out the stereovision.
The above-mentioned embodiments only represent some embodiments of the present invention, and the description thereof is specific and detailed, but not to be construed as limiting the scope of the present invention. It should be noted that, for those skilled in the art, without departing from the spirit of the present invention, several variations and modifications can be made, which are within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.
Claims (5)
1. A three-dimensional cell visualization device is characterized by comprising a spectroscope, an optical filter, a microscope, a charge-coupled device, a piezoelectric scanning controller and a processor;
the processor is respectively connected with the charge coupled device and the piezoelectric scanning controller;
the piezoelectric scanning controller is also connected with the spectroscope;
the spectroscope is used for decomposing exciting light and emitting light emitted by cells under the action of the exciting light, the optical filter is used for filtering the emitting light, and the charge-coupled device is used for collecting signals of the emitting light, obtaining a fluorescence image and sending the fluorescence image to the processor; the piezoelectric scanning controller controls the focal plane of the microscope to step along the optical axis to obtain a multilayer slice image of the cell image; and the processor performs deconvolution processing and stereoscopic visualization processing according to the fluorescence image and the multilayer slice image to obtain a three-dimensional image of the cell.
2. The device of claim 1, further comprising a light source for emitting excitation light, said light source being connected to said processor.
3. The cellular three-dimensional visualization device according to claim 1, wherein the beam splitter comprises a dichroic beam splitter.
4. The device of claim 1, wherein the microscope comprises a wide field fluorescence microscope.
5. The cellular three-dimensional visualization device according to claim 4, wherein the microscope structure comprises a slide and an objective lens.
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| CN202021553865.8U CN213022779U (en) | 2020-07-30 | 2020-07-30 | Three-dimensional cell visualization device |
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| CN202021553865.8U CN213022779U (en) | 2020-07-30 | 2020-07-30 | Three-dimensional cell visualization device |
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