CN102551661B - Fluorescence spectrum endoscopic imaging method and system - Google Patents
Fluorescence spectrum endoscopic imaging method and system Download PDFInfo
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- CN102551661B CN102551661B CN201010580863.2A CN201010580863A CN102551661B CN 102551661 B CN102551661 B CN 102551661B CN 201010580863 A CN201010580863 A CN 201010580863A CN 102551661 B CN102551661 B CN 102551661B
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Abstract
The invention provides a fluorescence spectrum endoscopic imaging method and a system, which are applicable to the field of photoelectric detection. The fluorescence spectrum endoscopic imaging method includes following steps: generating exciting light; dividing the exciting light into a plurality of sub light beams, leading the sub light beams to correspond to a plurality of sub areas of a sample and leading fluorescent materials to be distributed in the sample; adjusting the sub light beams and leading the sub light beams to be conducted into an organism and to be focused into the sub areas of the sample; scanning the sample by the aid of the sub light beams and leading the fluorescent materials in the sub areas to emit fluorescent light; and acquiring fluorescent light emitted during scanning in real time and generating a spectrum resolution fluorescent image. The sub light beams two-dimensionally scan the sample, so that the spectrum resolution fluorescent image of the total sample is acquired, time is short, speed is fast, injury to the organism is low, biomedical research, particularly early diagnosis of cancer, can be realized advantageously, and the fluorescence spectrum endoscopic imaging method and the system have an important significance.
Description
Technical field
The invention belongs to Photoelectric Detection field, relate in particular to a kind of fluorescence spectrum endoscopic imaging method and system.
Background technology
Fluorescence microscopy has become life sciences, especially the important tool of RESEARCH ON CELL-BIOLOGY.Excited Fluorescence Combined microtechnique has the lethal effect of life entity little, and penetration depth is large, has the advantages such as chromatography ability, has become the important means of life science.Fluorescence spectrum image can provide 26S Proteasome Structure and Function information for biomedical determination and analysis.
In recent years, along with the fast development of novel optical fiber and micro-fabrication technology, the research of optical fiber two-photon fluorescence microscope and endoscope makes the research of two-photon fluorescence micro-imaging technique in internal and the living animal of live body become possibility.In two-photon fluorescence spectrum, peep microtechnique at present and caused international great attention, make a large amount of achievements in research for this problem, obtained a lot of achievements in research at the aspect such as speck mirror and application thereof of endoscope system design, scan mechanism, optics conduction and high-NA.Be subject in vivo peeping application conditions restriction, imaging time is unsuitable long.But the speed of fluorescence spectrum based endoscopic imaging is slow at present, efficiency is low, and length consuming time causes great impact to organism.
Summary of the invention
The object of the embodiment of the present invention is to provide a kind of fluorescence spectrum endoscopic imaging method, is intended to solve slow, the inefficient problem of existing fluorescence spectrum based endoscopic imaging speed.
The embodiment of the present invention is achieved in that a kind of fluorescence spectrum endoscopic imaging method, comprises the following steps:
Produce exciting light;
Described exciting light is divided into multiple beamlets, and described multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in described sample;
Adjust described multiple beamlet, make each beamlet conduct in organism and focus on the subregion of described sample;
Utilize described multiple beamlet to scan described sample, make the fluorescent material in all subregion send fluorescence;
The fluorescence sending when Real-time Collection scanning, generates spectrally resolved fluoroscopic image.
Another object of the embodiment of the present invention is to provide a kind of fluorescent spectrum endoscope system, and described system comprises:
Excitation source, for generation of exciting light;
Beam splitter, for described exciting light is divided into multiple beamlets, described multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in described sample;
Flexible media, for adjusting described multiple beamlet, conducts in organism described multiple beamlet;
Concentrating element, for making each beamlet focus on the subregion of described sample;
Scanning element, for utilizing described multiple beamlet to scan described sample, makes the fluorescent material in all subregion send fluorescence;
Dichroic mirror and biography are as medium, for described fluorescence is derived in described organism;
Dispersion element, for making described fluorescence launch along spectrum direction;
Detector, the fluorescence sending while scanning for Real-time Collection, generates spectrally resolved fluoroscopic image;
Described dichroic mirror is located between described scanning element and concentrating element.
The embodiment of the present invention is divided into exciting light and the multiple subregions of sample multiple beamlets one to one, the plurality of beamlet is conducted in organism, each beamlet focuses on the subregion of sample, form multi-point shooting fluorescence, derive fluorescence and it is launched along spectrum direction, by multiple beamlets, sample is carried out to two-dimensional scan, thereby obtain the fluoroscopic image that whole sample spectra is differentiated, time is short, speed is fast, little to organism damage, be conducive to live body and study at body, particularly to early diagnosis of cancer, significant.
Brief description of the drawings
Fig. 1 is the realization flow figure of the fluorescence spectrum endoscopic imaging method that provides of the embodiment of the present invention;
Fig. 2 is structure and the index path thereof of the fluorescent spectrum endoscope system that provides of the embodiment of the present invention;
Fig. 3 a is scanning element the carries out line sweep location drawing to sample;
Fig. 3 b is the spectrally resolved fluoroscopic image corresponding with Fig. 3 a;
Fig. 4 a is scanning element carries out line sweep another location figure to sample;
Fig. 4 b is the spectrally resolved fluoroscopic image corresponding with Fig. 4 a.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
The embodiment of the present invention is divided into exciting light and the multiple subregions of sample multiple beamlets one to one, the plurality of beamlet is conducted in organism, each beamlet focuses on the subregion of sample, form multi-point shooting fluorescence, derive fluorescence and it is launched along spectrum direction, by multiple beamlets, sample is carried out to two-dimensional scan, thereby obtain the fluoroscopic image that whole sample spectra is differentiated, time is short, speed is fast, little to organism damage, is conducive to live body and studies at body.
The fluorescence spectrum endoscopic imaging method that the embodiment of the present invention provides comprises the following steps:
Produce exciting light;
Described exciting light is divided into multiple beamlets, and described multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in described sample;
Adjust described multiple beamlet, make each beamlet conduct in organism and focus on the subregion of described sample;
Utilize described multiple beamlet to scan described sample, make the fluorescent material in all subregion send fluorescence;
The fluorescence sending when Real-time Collection scanning, generates spectrally resolved fluoroscopic image.
The fluorescent spectrum endoscope system that the embodiment of the present invention provides comprises:
Excitation source, for generation of exciting light;
Beam splitter, for described exciting light is divided into multiple beamlets, described multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in described sample;
Flexible media, for adjusting described multiple beamlet, conducts in organism described multiple beamlet;
Concentrating element, for making each beamlet focus on the subregion of described sample;
Scanning element, for utilizing described multiple beamlet to scan described sample, makes the fluorescent material in all subregion send fluorescence;
Dichroic mirror and biography are as medium, for described fluorescence is derived in described organism;
Dispersion element, for making described fluorescence launch along spectrum direction;
Detector, the fluorescence sending while scanning for Real-time Collection, generates spectrally resolved fluoroscopic image;
Described dichroic mirror is located between described scanning element and concentrating element.
Below in conjunction with specific embodiment, realization of the present invention is described in detail.
Fig. 1 shows the realization flow of the fluorescence spectrum endoscopic imaging method that the embodiment of the present invention provides, and details are as follows:
In step S101, produce exciting light;
The preferred operating frequency of the embodiment of the present invention is 76MHz, and the cycle is 120fs, and femtosecond (ultrashort) pulse laser that centre wavelength is 800nm is as exciting light, and this exciting light can be realized the two-photon excitation of fluorescent material.Conventionally, paired pulses laser carries out beam-expanding collimation and adjusts its intensity distributions, forms the flat top beam that even intensity distributes.
In step S102, exciting light is divided into multiple beamlets, multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in sample;
The exciting light that the embodiment of the present invention distributes even intensity is divided into multiple beamlets, and the plurality of beamlet is one by one corresponding to multiple subregions of sample with fluorescent material.
In step S103, adjust multiple beamlets, make each beamlet conduct in organism and focus on the subregion of sample;
The embodiment of the present invention makes multiple beamlet parallel conductions to organism, focuses on separately the subregion of sample.Particularly, first make multiple beamlets be coupled into flexible media, multiple beamlets are via flexible media and be advanced in organism, and in organism, multiple beamlet line focuses form exciting light array points and are projected to respectively the subregion of correspondence with it.Wherein flexible media is photonic crystal fibre array, and its input is positioned in vitro, and outfan is positioned at organism.
Before multiple beamlets are coupled into flexible media, need collimate to each beamlet, make each beamlet become directional light.Multiple beamlets, after flexible media output, also need it to collimate, and are convenient to each beamlet and focus on corresponding with it sample subregion.
In step S104, utilize multiple beamlets to scan sample, make the fluorescent material in all subregion send fluorescence;
Scanning is divided into line sweep and step-scan by the embodiment of the present invention, and detailed process is as follows:
1, line sweep
Multiple beamlet line focuses form exciting light array point and are projected to sample, longitudinally all subregion are carried out to line sweep along sample, and the fluorescent material in all subregion sends fluorescence under the effect of exciting light array point.The speed of this line sweep is fast, the time is short.
2, step-scan
After the longitudinal line sweep of each sub regions is finished, laterally each sub regions is carried out to step-scan along sample and adjust exciting light array point in the horizontal position of sample.
Above-mentioned line sweep and step-scan are carried out in circulation, until the scanning of the each sub regions of complete paired samples.The direction that can also exchange line sweep and step-scan while should be appreciated that concrete enforcement.
In step S105, the fluorescence sending when Real-time Collection scanning, generates spectrally resolved fluoroscopic image.
When the embodiment of the present invention scans sample all subregion, gather the fluorescence that in all subregion, fluorescent material sends, generate spectrally resolved fluoroscopic image.Particularly, first derive the fluorescence that in all subregion, fluorescent material sends, then fluorescence is launched along its spectrum direction, then gather spectrally resolved fluorescence information, generate fluoroscopic image.
Those of ordinary skill in the art is to be understood that, realizing all or part of step in above-described embodiment method can carry out the hardware that instruction is relevant by program and complete, this program can be stored in a computer read/write memory medium, as ROM/RAM, disk, CD etc.
Fig. 2 shows the structure of the fluorescent spectrum endoscope system that the embodiment of the present invention provides, and for convenience of explanation, only shows the part relevant to the embodiment of the present invention.
The fluorescent spectrum endoscope system that the embodiment of the present invention provides has an excitation light path and and surveys light path.Excitation light path comprises excitation source, beam-expanding collimation device, reshaper, beam splitter, collimating lens, the first coupled lens, photonic crystal fibre array, the first GRIN Lens, scanning element and speck mirror.Survey light path and comprise speck mirror, dichroic mirror, the second GRIN Lens, image-carrying fiber bundle, the second coupled lens, filter element, correction element, dispersion element, imaging len and detector.Wherein speck mirror by excitation light path and survey light path shared.
Below the structure of excitation light path is elaborated.
As shown in Figure 2, the preferred titanium gem of embodiment of the present invention femto-second laser 1 is as excitation source, and it can produce centre wavelength is that 800nm, frequency are 76MHz, the cycle pulse laser that is 120fs, and this pulse laser can be realized the two-photon excitation of fluorescent material.Pulse laser becomes the collimated light of required size via beam-expanding collimation device 2.
In the embodiment of the present invention, reshaper is beam shaping 3, and the pulse laser of collimation, through beam shaping 3 shapings, forms the flat top beam that even intensity distributes.Beam splitter can be microlens array, diffraction optical element or beam splitter, the preferred microlens array 4 of the present embodiment, the pulse laser that flat-top distributes is divided into multiple beamlets through microlens array 4, multiple subregions of multiple beamlet counter samples 12, in the present embodiment, microlens array 4 is that 3 × 3 microlens arrays are that microlens array has nine speck mirrors.
Wherein the back focal plane of collimating lens 5 overlaps with the front focal plane of microlens array 4, and beamlet focuses at the back focal plane of collimating lens 5 at the front focal plane of microlens array 4, and each beamlet all becomes the beamlet of directional light through collimating lens 5.
Above-mentioned multiple beamlet focuses on and is coupled into photonic crystal fibre array 7 through the first coupled lens 6.Photonic crystal fibre array 7 is equidistantly arranged and is formed by many photonic crystal fibers, and the number of photonic crystal fiber and arrangement mode thereof are identical with microlens array 4.Multiple beamlets conduct in organism through photonic crystal fibre array 7, are projected to scanning element 9 from multiple beamlets of photonic crystal fibre array 7 outgoing through the first GRIN Lens 8.GRIN Lens is the radially excellent lens of gradual change of refractive index, and each beamlet all becomes directional light through the first GRIN Lens 8.Each beamlet is projected to the sample 12 with fluorescent material through scanning element 9, be provided with the speck mirror 11 of converging action between sample 12 and scanning element 9.Described scanning element 9 is preferably MEMS (Micro-Electro-Mechanical Systems, MEMS) scanning mirror, and MEMS scanning mirror is two-dimensional scan mirror, can carry out line sweep and step-scan to sample.Multiple beamlets carry out two-dimensional scan through scanning element 9 to sample, and detailed process is as follows:
1, line sweep
Multiple beamlets focus on formation exciting light array point through speck mirror 11 and are projected to sample, longitudinally all subregion is carried out to line sweep along sample 12, fluorescent material in all subregion sends fluorescence and forms phosphor dot array under the effect of exciting light array point, forms fluorescence linear array through scanning element 9 line sweeps.The speed of this line sweep is fast, the time is short.
2, step-scan
After the longitudinal line sweep of each sub regions is finished, laterally each sub regions is carried out to step-scan along sample and adjust exciting light array point in the horizontal position of sample.
Above-mentioned line sweep and step-scan are carried out in circulation, until the scanning of the each sub regions of complete paired samples.The direction that can also exchange line sweep and step-scan while should be appreciated that concrete enforcement.
Each beamlet forms exciting light array point and is projeced into the subregion of sample 12 after this excitation light path conduction, and the fluorescent material in excited sample 12 sends fluorescence.Correspondingly, this fluorescence also has multiple beamlets.When scanning element 9 carries out line sweep, gather spectrally resolved fluorescence information by detector 20, generate fluoroscopic image.This spectrally resolved fluorescence information comprises strength information, spectral information and the positional information in sample thereof of fluorescence.
Below the structure of surveying light path is elaborated.
The embodiment of the present invention is surveyed dichroic mirror 10 in light path and is located between scanning element 9 and speck mirror 11, the pulse laser that dichroic mirror 10 is 800nm to centre wavelength is thoroughly high, the fluorescence that is 400～700nm to wavelength is high anti-, and the angle between dichroic mirror 10 and fluorescence is 45 ° or 135 °.Above-mentioned fluorescence is collected by speck mirror 11, forms the fluorescence beamlet of multi-beam collimation, and dichroic mirror 10 reflects each fluorescence beamlet from excitation light path, focus on body the inner of image-carrying fiber bundle 14 through the second GRIN Lens 13.Each fluorescence beamlet is derived in organism by image-carrying fiber bundle 14, change into multi-path parallel light through the second coupled lens 13, by filter element 16 filtering exciting lights and other veiling glare, be projected to dispersion element 18 through correction element 17 (as scanning mirror), dispersion element 18 launches to form spectrally resolved multi-thread array by multi beam fluorescence beamlet along spectrum direction, focused on again the sensitive area of detector 20 by imaging len 19, the fluorescence intensity information of the multi-thread array that spectra re-recorded is differentiated, generates spectrally resolved fluoroscopic image.
The wherein body inner face conjugate planes each other of the focal plane of the interior exciting light array point of sample 12 and image-carrying fiber bundle 14, the sensitive area conjugate planes each other of the external end face of image-carrying fiber bundle 14 and detector 20, that is to say that the phosphor dot array inspiring from sample 12 diverse locations focuses on the correspondence position of image-carrying fiber bundle 14 body inner faces through speck mirror 11 and the second GRIN Lens 13, reach the correspondence position of its external end face through image-carrying fiber bundle 14, by dispersion element 18, phosphor dot array being launched to form spectrally resolved fluorescence intensity along spectrum direction distributes, focused on again the correspondence position of detector 20 by imaging len 19.The fluorescence that detector 20 can send sample 12 like this carries out spectrographic detection, generates spectrally resolved fluoroscopic image.
In the embodiment of the present invention, detector 20 is that planar array detector is preferably CCD camera or CMOS camera for generating the planar array detector of spectrally resolved fluoroscopic image.Planar array detector is connected with computer 21, the spectrally resolved fluoroscopic image that computer 21 is surveyed for storing, process and read detector 20, and by the exposure of computer 21 chain of command array detectors.
Above-mentioned spectrum direction is vertical with the scanning direction of scanning element 9, if scanning direction is directions X, spectrum direction should be Y-direction.Scanning element 9 line sweep at the beginning, detector 20 starts exposure; Scanning element 9 line sweeps make sample 12 produce fluorescence linear array; When scanning element 9 completes primary line scanning to sample 12, detector 20 single exposures finish, and now record a spectrally resolved fluoroscopic image, and this fluoroscopic image is corresponding to many line positions of sample, detector 20 has recorded the information of more than 12 photoluminescence line of sample thus, as shown in Figure 3 a and Figure 3 b shows.Afterwards, scanning element (MEMS scanning mirror) 9 is along pixel of other direction Y+ stepping, and correction element 17, correspondingly along pixel of Y-direction stepping, makes fluorescence linear array in the invariant position of detector 20.Then, scanning element (MEMS scanning mirror) 9 starts to repeat above-mentioned line sweep process, detector 20 also starts to repeat above-mentioned exposure process, 9 pairs of sample 12 line sweeps of scanning element (MEMS scanning mirror) complete, detector 20 end exposures simultaneously, record another spectrally resolved fluoroscopic image, this image is corresponding to many line positions of another group in sample 12, as shown in Fig. 4 a and Fig. 4 b.Repeat said process, until complete sample 12 of exciting light array spot scan.So far, obtained sample 12 interior somewhat spectrally resolved fluoroscopic image, can obtain the spectrally resolved fluorescence information of sample 12 by algorithm reconstruct.
When scanning element (MEMS scanning mirror) 9 is during along a pixel of other direction Y+ stepping, can make detector 20 correspondingly along pixel of Y-direction stepping, so that fluorescence linear array is in the invariant position of detector 20, can save correction element 17, simplify fluorescent spectrum endoscope system, improve precision, cost-saving.
The embodiment of the present invention is divided into exciting light and the multiple subregions of sample multiple beamlets one to one, the plurality of beamlet is conducted in organism, each beamlet focuses on the subregion of sample, form multi-point shooting fluorescence, derive fluorescence and it is launched along spectrum direction, by multiple beamlets, sample is carried out to two-dimensional scan, thereby obtain the fluoroscopic image that whole sample spectra is differentiated, time is short, speed is fast, little to organism damage, be conducive to live body and study at body, particularly to early diagnosis of cancer, significant.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. a fluorescence spectrum endoscopic imaging method, is characterized in that, said method comprising the steps of:
Produce exciting light;
Described exciting light is divided into multiple beamlets, and described multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in described sample;
Adjust described multiple beamlet, make each beamlet conduct in organism and focus on the subregion of described sample;
Utilize described multiple beamlet to scan described sample, make the fluorescent material in all subregion send fluorescence, wherein each beamlet scans described sample through MEMS scanning mirror;
The fluorescence sending when Real-time Collection scanning, generate spectrally resolved fluoroscopic image, wherein said fluorescence is first collected by speck mirror, form the fluorescence beamlet of multi-beam collimation, by dichroic mirror, each fluorescence beamlet is reflected from excitation light path again, focus on body the inner of image-carrying fiber bundle through the second GRIN Lens, the focal plane of exciting light array point and the body inner face conjugate planes each other of image-carrying fiber bundle in described sample, the sensitive area conjugate planes each other of the external end face of described image-carrying fiber bundle and detector; Wherein, the described multiple beamlets of described adjustment, make each beamlet conduct in organism and the step that focuses on the subregion of described sample is specially:
Described multiple beamlet is coupled into photonic crystal fibre array, enter in organism via described photonic crystal fibre array, be projected to scanning element from multiple beamlets of described photonic crystal fibre array outgoing through the first GRIN Lens, each beamlet all becomes directional light through described the first GRIN Lens, and described photonic crystal fibre array is equidistantly arranged and formed by many photonic crystal fibers;
In described organism, each directional light focuses on formation exciting light array point through speck mirror and is projected to described sample.
2. fluorescence spectrum endoscopic imaging method as claimed in claim 1, it is characterized in that, described scanning is divided into line sweep and step-scan, describedly utilizes described multiple beamlet to scan described sample, and the step that makes fluorescent material in all subregion send fluorescence is specially:
Each beamlet scans corresponding subregion along described sample vertical line, makes the fluorescent material in all subregion send fluorescence;
After described line sweep finishes, laterally corresponding subregion is carried out to step-scan along described sample, adjust each beamlet in the horizontal position of described sample;
Described line sweep and step-scan are carried out in circulation, until complete the scanning to each sub regions.
3. fluorescence spectrum endoscopic imaging method as claimed in claim 1, is characterized in that, the fluorescence sending when described Real-time Collection scanning, and the step that generates spectrally resolved fluoroscopic image is specially:
Derive the fluorescence that in all subregion, fluorescent material sends;
Described fluorescence is launched along spectrum direction;
The fluorescence information that Real-time Collection is spectrally resolved, generates fluoroscopic image.
4. fluorescence spectrum endoscopic imaging method as claimed in claim 1, is characterized in that, further comprising the steps of after the step of described generation exciting light:
Described exciting light is carried out to beam-expanding collimation;
Adjust the intensity distributions of described exciting light, make the intensity distributions of described exciting light even;
The fluorescence sending when described Real-time Collection scanning, the step that generates spectrally resolved fluoroscopic image is before further comprising the steps of:
Adjust described fluorescence, make described fluorescence in the invariant position of detector.
5. a fluorescent spectrum endoscope system, is characterized in that, described system comprises:
Excitation source, for generation of exciting light;
Beam splitter, for described exciting light is divided into multiple beamlets, described multiple beamlets, corresponding to multiple subregions of sample, are distributed with fluorescent material in described sample;
Flexible media, for adjusting described multiple beamlet, conducts in organism described multiple beamlet;
Concentrating element, for making each beamlet focus on the subregion of described sample;
Scanning element, for utilizing described multiple beamlet to scan described sample, makes the fluorescent material in all subregion send fluorescence;
Dichroic mirror and biography are as medium, for described fluorescence is derived in described organism;
Dispersion element, for making described fluorescence launch along spectrum direction;
Detector, the fluorescence sending while scanning for Real-time Collection, generates spectrally resolved fluoroscopic image;
Described dichroic mirror is located between described scanning element and concentrating element, and the described flexible media many photonic crystal fibers of serving as reasons are equidistantly arranged the photonic crystal fibre array of formation.
6. fluorescent spectrum endoscope system as claimed in claim 5, is characterized in that, between described excitation source and described beam splitter, is also provided with:
Beam-expanding collimation device, for adjusting the size of described exciting light and collimating;
Reshaper, for adjusting the intensity distributions of described exciting light, makes the intensity distributions of described exciting light even;
Between described beam splitter and described flexible media, be also provided with:
Collimating lens, for collimating each beamlet, makes each beamlet become directional light;
The first coupled lens, for making each beamlet be coupled into described flexible media;
Between described flexible media and described scanning element, be also provided with:
The first GRIN Lens, for collimating from each beamlet of described flexible media output;
Described dichroic mirror and described biography are as being also provided with between medium:
The second GRIN Lens, for making described fluorescence focus on described biography body the inner as medium;
Described biography is as being also provided with between medium and described dispersion element:
The second coupled lens, for collimating the fluorescence as medium output from described biography;
Correction element, for adjusting described fluorescence, makes described fluorescence in the invariant position of described detector;
Imaging len, for by described fluorescence imaging in described detector;
Described concentrating element is speck mirror.
7. fluorescent spectrum endoscope system as claimed in claim 6, it is characterized in that, described beam splitter is microlens array, diffraction optical element or beam splitter, and the front focal plane of described microlens array overlaps with the back focal plane of described collimating lens, and described biography is image-carrying fiber bundle as medium.
8. fluorescent spectrum endoscope system as claimed in claim 7, is characterized in that, described fluorescence is collected by described speck mirror, and forms the fluorescence beamlet of multi-beam collimation; Described dichroic mirror reflects described fluorescence beamlet from excitation light path, focus on body the inner of described image-carrying fiber bundle through described the second GRIN Lens.
9. fluorescent spectrum endoscope system as claimed in claim 8, it is characterized in that, the focal plane of exciting light array point and the external end face conjugate planes each other of described image-carrying fiber bundle in described sample, the sensitive area conjugate planes each other of the external end face of described image-carrying fiber bundle and described detector.
10. the fluorescent spectrum endoscope system as described in any one in claim 5～9, is characterized in that, described scanning element line sweep at the beginning, and described detector starts exposure; When described scanning element completes primary line scanning to described sample, described detector single exposure finishes; The time that described detector exposes is once identical with the described scanning element line sweep time once.
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| CN113469864B (en) * | 2021-06-28 | 2024-05-07 | 平湖莱顿光学仪器制造有限公司 | Method and equipment for acquiring microscopic image |
| CN115702777B (en) * | 2021-08-13 | 2024-05-17 | 深圳先进技术研究院 | Multifunctional two-photon microscopic imaging system |
| CN115553724A (en) * | 2022-09-27 | 2023-01-03 | 宁夏祥瑞智能科技有限公司 | Portable single photon calcium imaging microscopic equipment and method |
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