CN113916848A - Method and system for generating light beam through stimulated radiation loss imaging of strong scattering medium - Google Patents
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Abstract
The utility model provides a light beam generation method and system through stimulated radiation loss imaging of strong scattering medium, comprising the following steps: acquiring an excitation beam and a loss beam; obtaining a combined wavefront of the shaped excitation light beam and the shaped loss light beam according to the obtained excitation light beam, the obtained loss light beam and the transmission matrix; generating a light beam according to the shaped combined wavefront and the strong scattering medium; the combined wave front of the shaped excitation beam and the loss beam uses a transmission matrix and a multiplexing computation holographic technology to realize the overlapping of the excitation beam and the loss beam on time and space. Based on the dual-wavelength transmission matrix, the excitation light and the loss light simultaneously generate the excitation light beam and the loss light beam which are overlapped in time and space after passing through the strong scattering medium, and the rapid scanning of the light beams is realized.
Description
Technical Field
The disclosure belongs to the technical field of optical microscopic imaging, and particularly relates to a method and a system for generating a light beam through stimulated radiation loss imaging of a strong scattering medium.
Background
The statements in this section merely provide background information related to the present disclosure and may not necessarily constitute prior art.
The Stimulated radiation loss (STED) microscope has super-resolution imaging capability, can expand a visual field to a nanometer level, breaks through the limit of a diffraction limit on the resolution of an optical microscope, realizes super-resolution microscopic imaging, and is widely applied to biological imaging.
Compared with the confocal imaging technology, the STED microscopic imaging technology utilizes a pair of focuses overlapped in space and time to form a laser illumination beam, and the ingenious design is the key for realizing super-resolution imaging. One focus is a Gaussian focus point serving as an excitation light beam and used for realizing excitation of fluorescent molecules; the other focus is the doughnut beam, which is called the STED beam, for stimulated emission depleted fluorescence. Although both laser foci are diffraction limited, subtracting the latter from the former produces a sub-diffracted fluorescence focus, which is scanned across the sample to obtain an image of the sub-diffracted order. Therefore, the primary condition to achieve STED imaging is to generate the illumination beam it requires. Since the invention of the STED microscope, it was generally used for super-resolution imaging of samples in an environment with a uniform refractive index distribution outside an animal, and since the environment inside an animal living body is complex, in vitro experiments cannot exactly simulate the environment inside the animal living body. However, when the STED imaging of the inside of an animal living body is to be performed, because a strong scattering medium such as a biological tissue disturbs the wavefront of an incident light beam, the formation of an excitation focus and an STED focus is suppressed, and thereby the STED imaging cannot be performed. When the structured illumination beam required in STED imaging passes through a strongly scattering medium, such as biological tissue, multiple scattering of light in the medium tears the wavefront of the structured illumination beam, thereby inhibiting STED imaging.
Currently, there have been many efforts to achieve STED imaging through heterogeneous media. For a specimen with weak light disturbance, such as a retina slice or a fruit fly brain, the wave front is disturbed less, and generally inclines, defocuses and the like, and the factors only introduce small deviation to the formation of a focus point. In these cases, aberration correction for two focal points in the STED microscope can be achieved using a zernike polynomial correction method in adaptive optics. To increase the imaging depth of the STED, on the one hand, two-photon excitation techniques can be introduced into the STED microscope, since the photons used in these experiments have a longer wavelength, usually in the near infrared band, which can penetrate deeper scattering tissues. On the other hand, an optical needle having a self-repair capability may also be used for excitation and depletion of fluorescence. However, when it is desired to apply STED imaging behind or inside a strongly scattering medium, for example: thick biological tissue, the above methods all fail. Under strongly scattering medium conditions, multiple scattering will disturb the wavefront of the incident excitation and STED beams, producing a random intensity speckle pattern. In fact, multiple scattering can be seen as a linear and deterministic process, in which the input field and the scattered field can be correlated by a Transmission Matrix (TM). Based on the transmission matrix, a variety of techniques have been proposed to achieve wavefront modulation through strongly scattering media. These techniques pave the way for microscopic imaging in deep tissues. However, the illumination beam required in generating STED imaging after a strongly scattering medium has not been investigated.
Disclosure of Invention
In order to solve the above problems, the present disclosure provides a method and a system for generating a light beam through stimulated radiation loss imaging of a strong scattering medium, based on a dual-wavelength transmission matrix, so that an excitation light beam and a loss light beam which are overlapped in time and space are simultaneously generated after the excitation light and the loss light pass through the strong scattering medium, thereby realizing rapid scanning of the light beam.
According to some embodiments, a first aspect of the present disclosure provides a method for generating a light beam through stimulated radiation loss imaging of a strong scattering medium, which adopts the following technical solutions:
a method of generating a beam for stimulated radiation depletion imaging through a strongly scattering medium, comprising the steps of:
acquiring an excitation beam and a loss beam;
obtaining a combined wavefront of the shaped excitation light beam and the shaped loss light beam according to the obtained excitation light beam, the obtained loss light beam and the transmission matrix;
generating a light beam according to the shaped combined wavefront and the strong scattering medium;
the combined wave front of the shaped excitation beam and the loss beam uses a transmission matrix and a multiplexing computation holographic technology to realize the overlapping of the excitation beam and the loss beam on time and space.
As a further technical limitation, after the excitation and loss beams are acquired, calibration of the excitation and loss beam transmission matrices is required.
Further, there is no correlation between the excitation beam transmission matrix and the loss beam transmission matrix, and both transmission matrices can be measured separately.
Further, the measured excitation beam transmission matrix is subjected to optical phase conjugation operation to obtain a conjugate wavefront, and a required excitation focus is formed on an output plane according to the obtained conjugate wavefront.
Further, point spread function modulation is carried out on the measured loss light beam, and a loss focus in point spread distribution is obtained.
Further, a spatial light modulator is used for loading a multiplexing calculation hologram, wave front shaping is carried out on the excitation light beam and the loss light beam at the same time, the shaped combined wave front is incident on a strong scattering medium, an overlapping focus is constructed, and the light beam is generated.
According to some embodiments, a second aspect of the present disclosure provides a light beam generating system for stimulated radiation loss imaging through a strong scattering medium, which adopts the following technical solutions:
a beam generation system for stimulated radiation depletion imaging through a strongly scattering medium, comprising:
a laser for acquiring an excitation beam and a loss beam;
the light beam processing module adjusts the focus of the excitation light beam and the focus of the loss light beam to be overlapped on time and space based on the transmission matrix and the multiplexing calculation holographic technology, and performs wave front shaping on the laser light beam and the loss light beam to obtain the combined wave front of the shaped excitation light beam and the loss light beam;
and the light beam generation module is used for transmitting the shaped combined wavefront through a strong scattering medium to generate a light beam.
As a further technical limitation, a beam splitter and a beam expander are sequentially arranged between the laser and the beam processing module; and the excitation light beam and the loss light beam both adopt continuous light sources.
As a further technical limitation, the beam processing module adopts a digital micro-mirror device, and comprises a beam calibration module, a beam space processing unit and a beam time processing unit.
Further, the light beam calibration module performs optical phase conjugation operation on the acquired excitation light beam through a transmission matrix to obtain an excitation focusing light beam, and performs point spread function modulation on the acquired loss light beam through the transmission matrix to obtain a loss focusing light beam.
Compared with the prior art, the beneficial effect of this disclosure is:
the present disclosure utilizes multiplexed holographic techniques to construct overlapping focal points with fine structures for use in 2D STED and 3D STED microscopes. In addition, under the condition of no mechanical motion, based on the rapid switching capability of the digital micro-mirror device, the rapid scanning of the illumination light beam can be rapidly realized, and a way is opened for STED imaging behind a strong scattering medium (such as thick biological tissue).
Drawings
The accompanying drawings, which are included to provide a further understanding of the disclosure, illustrate embodiments of the disclosure and together with the description serve to explain the disclosure and are not to limit the disclosure.
FIG. 1 is a flow chart of a method for generating a beam through stimulated radiation depletion imaging of a strongly scattering medium according to an embodiment of the disclosure;
FIG. 2(a) is a wavefront plot of a strongly scattering medium perturbing the excitation and STED beams in one embodiment of the present disclosure;
FIG. 2(b) is a diagram of a process of performing wavefront modulation through a strong scattering medium based on a dual-wavelength transmission matrix according to an embodiment of the disclosure;
FIG. 2(c) is a diagram of a focal point of an overlap generated by a multiplexing holography technique through a strong scattering medium according to an embodiment of the present disclosure;
FIG. 3 is a graph of an illumination beam for generating a 2D STED image through a ZnO scattering layer according to a first embodiment of the disclosure;
FIG. 4 is a diagram of an illumination beam for generating a 3D STED image through a ZnO scattering layer according to a first embodiment of the disclosure;
FIG. 5 is a block diagram of a beam generation system for stimulated radiation depletion imaging through a strong scattering medium according to a second embodiment of the disclosure;
fig. 6 is an experimental design diagram of a beam generation system for stimulated radiation depletion imaging through a strong scattering medium in the second embodiment of the disclosure.
The specific implementation mode is as follows:
the present disclosure is further described with reference to the following drawings and examples.
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
The embodiments and features of the embodiments in the present disclosure may be combined with each other without conflict.
Example one
The embodiment of the disclosure introduces a method for generating a light beam through stimulated radiation loss imaging of a strong scattering medium.
The embodiment introduces a technology for realizing an illumination light beam required by STED microscopic imaging through a strong scattering medium structure, and utilizes a method of a dual-wavelength transmission matrix to simultaneously generate an excitation light beam and a loss light beam after the excitation light beam and the loss light beam pass through the strong scattering medium, and the two light beams are overlapped in time and space; the overlapping focal points are generated by using a Digital Micromirror Device (DMD for short). Generating an illumination beam of STED by a strongly scattering medium would be advantageous for application of STED microscopy in deep tissue.
A method for generating a beam for stimulated radiation depletion imaging through a strongly scattering medium as shown in fig. 1, comprising the steps of:
acquiring an excitation beam and a loss beam;
obtaining a combined wavefront of the shaped excitation light beam and the shaped loss light beam according to the obtained excitation light beam, the obtained loss light beam and the transmission matrix;
generating a light beam according to the shaped combined wavefront and the strong scattering medium;
the combined wave front of the shaped excitation beam and the loss beam uses a transmission matrix and a multiplexing computation holographic technology to realize the overlapping of the excitation beam and the loss beam on time and space.
In this embodiment, the calibration work is first performed on the two-wavelength transmission matrices of the excitation and STED beams. In order to generate the excitation beam, an Optical Phase Conjugation (OPC) operation is performed on the transmission matrix of the excitation beam, and an excitation beam focusing point can be generated behind the scattering medium. For the STED light beam, a Point Spread Function (PSF) modulation method based on a transmission matrix is adopted to generate two-dimensional and three-dimensional STED focused light beams through a strong scattering medium. In a STED microscope, the excitation focus and the STED focus need to be strictly overlapping in time and space. To meet this need, the wavefronts of the excitation and STED beams are simultaneously modulated using multiplexed computational holography, the excitation and STED foci are generated simultaneously through a strongly scattering medium and transmitted coaxially, and rapid wavefront shaping is achieved using a DMD.
When the excitation and STED beams are incident on a strong scattering medium, the wavefronts of the two beams are disturbed, producing speckle of random intensity behind the scattering medium, as shown in fig. 2 (a). In this case, the effect of the excitation beam and the STED beam is completely suppressed. Since the process of scattering is linear and deterministic, the TM approach is used to control the output light field through the scattering medium. From the mathematical description of wavefront shaping, it is assumed that the light fields at the input and output planes are N × 1 and M × 1 vectors, respectively, where N and M are expressed as input and output mode numbers, respectively. According to TM theory, although the light field is highly chaotic during multiple scattering, the output field (u) iso) And an input field (u)i) The relationship between them follows a linear equation: u. ofo＝TλuiWherein, TλRepresenting the transmission matrix (M x N matrix) of light of wavelength lambda through the scattering medium. When the exciting light and the loss light pass through the same strong scattering medium, their transmission matrixes are respectively TgAnd TrTo describe. Typically, in STED imaging, the wavelengths of the excitation light and the STED light differ by about 100 nm, and there is no correlation between the transmission matrices of the two wavelengths after they pass through a strong scattering medium. Thus, the transmission matrices of the two beams can be measured separately.
In the present embodiment, the transmission matrix is measured by the method described in the documents S.Popoff, G.Lerosey, R.Carnation, M.Fink, A.Boccara, and S.Gigan, "Measuring the transmission matrix in optics, an ap approach to the study and control of light propagation in distributed media," Physical review letters 104, 1002010 (601).
After the two transmission matrices are accurately measured, the corresponding wavefronts when the two light beams pass through the strong scattering medium to obtain the required light intensity distribution on the output plane can be calculated, and the detailed process is shown in fig. 2 (b).
As shown in (b) of FIG. 2(c), for the measured TgPerforming OPC operation (selecting to generate focus point on the most intermediate output mode to obtain a conjugate wavefront, and inputting the conjugate wavefrontOnto a strongly scattering medium, the desired excitation focus can be formed on the output plane. To generate the STED focus (donut or light bottle beam), a TM-based PSF modulation method is employed to obtain the PSF profile of the desired focus. According to the method, the PSF distribution in the scattering medium system can be modulated by performing numerical filtering on a virtual Fourier plane corresponding to the TM output mode by using a designed Mask. Mask used as filtering corresponds to the fourier transform of the PSF distribution. For a 2D STED focus, its Mask distribution is a vortex phase, and for a 3D STED focus, its Mask is a pi-step phase.
T measured as shown in the process of 2(b)rAfter Mask digital filtering, filtered T is obtainedr. Further, the filtered T is filteredrAfter performing the OPC operation (or selecting to generate a focus spot on the most intermediate output mode), a "donut" or "bottle" intensity distribution is obtained at the focal plane. The STED focus in fig. 2(b) shows the donut-shaped intensity distribution of the STED beam in 2D STED imaging.
In STED imaging, it is crucial that the excitation focus and the STED focus are coaxial. For the purpose of overlapping the two focal points spatially, the measured TgAnd filtered TrWhen performing the corresponding OPC operation, the same output mode is selected. In order to make the two foci overlap in time, it is proposed that a multiplexed computational holography technique can be used. Finally, a Spatial Light Modulator (SLM) is used to load the multiplexing calculation hologram, and the two beams of Light are simultaneously subjected to wavefront shaping, so that when the shaped combined wavefront is incident on the strong scattering medium, an overlapping focus can be constructed behind the strong scattering medium, as shown in fig. 2 (c).
T after calibration of ZnO scattering layergAnd TrThen, the illumination beam in 2D STED imaging is first constructed behind the ZnO scattering layer. First, for TgThe OPC operation was performed to generate a conjugate wavefront that transmitted through the ZnO to excite the focus spot, and then the Lee method (in this example, documents X.Hu, Q.ZHao, P.Yu, X.Li, Z.Wang, Y.Li, and L.Gong, "Dynamic profiling of organic-angular-particles for information) was usedthe method described in "Opt Express 26,1796-1808 (2018)") encodes this conjugate wavefront, and the resulting binarized amplitude hologram is shown in fig. 3(a) (note: only a portion of the multiplexed computed hologram is shown here for clarity). After loading this hologram on the DMD, a tightly focused excitation focus is created in the middle of the output plane through the ZnO, as shown in fig. 3 (d). Then, by measuring TrVortex phase filtering is carried out to obtain filtered TrThen, the OPC operation is performed on the encoded binary amplitude map, and the corresponding encoded binary amplitude map is shown in fig. 3 (b). Loading the binary amplitude diagram on a DMD, and generating a doughnut-shaped STED focus at the middle position of an output plane through a ZnO scattering layer, wherein the light intensity distribution is shown as a graph in FIG. 3 (e); obtaining a doughnut-shaped intensity distribution, wherein the intensity at the center of the doughnut is almost zero, and the periphery of the doughnut has higher intensity distribution; a plurality of two-dimensional intensity maps are then captured along the z-axis with a movable CMOS camera and processed to obtain x-z intensity profiles of the beam propagation, as shown in fig. 3(g) and 3(h), respectively. To generate both the excitation and depletion foci, FIGS. 3(a) and 3(b) are superimposed to obtain a multiplexed computed hologram, as shown in FIG. 3 (c). This hologram is loaded on the DMD, and both the excitation focus and the STED focus are generated through the ZnO scattering layer. The intensity distribution and the x-z intensity profile at the output plane are shown in fig. 3(f) and 3(i), respectively, and it can be seen that the two foci are well aligned. The intensity distribution along the cross-section of the white dashed line in fig. 3(i) is shown in fig. 3(j), and it can be seen from both curves that the two focal points coincide well. In FIG. 3, (a-c) are used to generate a binary amplitude map corresponding to the excitation beam, the doughnut-shaped STED beam, and the superimposed beam of the two; for clarity, only a portion of the amplitude map is presented here; (d-f) generating an intensity map of the excitation beam, the doughnut-shaped STED beam, and a superimposed beam of both beams in the x-y output plane; (g-i) an x-z plane intensity profile corresponding to the propagation of the light beam in (d-f); (j) light intensity distribution plot along the white dotted line in graph (f), round dot and square dot being fitted curves to the experimental data for excitation focus and STED focus, respectively。
Since the donut beam consumes only fluorescence in the x-y plane, a light bottle beam is typically used in order to limit the excitation focus in the z-axis direction. Thus, the illumination beam used in the 3D STED microscope is generated behind a strongly scattering medium. T measured by pi-step phase as Mask pairrAfter filtering and loading the multiplexed hologram on the DMD, an overlapping illumination beam is generated. The light intensity distribution in the x-y plane and the propagating light intensity distribution in the x-z plane of the overlapping focal points as shown in fig. 4(a) and 4(b) generate one bottle-shaped 3D STED focal point, and the two light beams overlap well; fig. 4(c) and 4(d) show beam intensity profiles along the horizontal direction broken line in fig. 4(b) and the vertical direction broken line in fig. 4(b), respectively. It can be seen that for the STED focus, the intensity null surrounds the high intensity region, and the excitation focus is centered as expected at the light bottle beam intensity null.
This embodiment utilizes multiplexed holographic techniques to construct overlapping focal points with fine structures for use in 2D STED and 3D STED microscopes. In addition, under the condition of no mechanical motion, based on the rapid switching capability of the digital micro-mirror device, the rapid scanning of the illumination light beam can be rapidly realized, and a way is opened for STED imaging behind a strong scattering medium (such as thick biological tissue).
Example two
The second embodiment of the present disclosure introduces a beam generation system that transmits stimulated radiation depletion imaging of a strong scattering medium, and adopts the beam generation method that transmits stimulated radiation depletion imaging of a strong scattering medium described in the first embodiment.
A beam generation system for stimulated radiation depletion imaging through a strongly scattering medium, as shown in fig. 5, comprising:
a laser for acquiring an excitation beam and a loss beam;
the light beam processing module adjusts the focus of the excitation light beam and the focus of the loss light beam to be overlapped on time and space based on the transmission matrix and the multiplexing calculation holographic technology, and performs wave front shaping on the laser light beam and the loss light beam to obtain the combined wave front of the shaped excitation light beam and the loss light beam;
and the light beam generation module is used for transmitting the shaped combined wavefront through a strong scattering medium to generate a light beam.
Fig. 6(a) is an experimental apparatus diagram, fig. 6(b) is a diagram of a primary light coincidence apparatus of two light beams, fig. 6(c) is a diagram of a calibration block required for measuring a transmission matrix, BS denotes a beam splitter, BE denotes a beam expander, M denotes a mirror, DMD denotes a digital micromirror device, L denotes a convex lens, ObJ denotes an objective lens, HSM denotes a strong scattering medium, Ref denotes a reference beam, and CMOS denotes a CMOS complementary metal oxide semiconductor camera.
In this example, a DMD (Vialux V7001) with a switching rate of up to 22.727kHz was used to achieve fast wavefront shaping, and the experimental setup schematic is shown in fig. 6 (a). To confirm the effectiveness of the method in example one, a laser having a wavelength of 532nm (Coblot Series04-01) and a laser having a wavelength of 633nm (HNL-210L, Thorlabs) were used as the excitation light and the loss light, respectively. The two beams are first combined at BS1 and then expanded to flood the surface of the DMD. Under the combined action of the 4f system and the spatial filter, the complex amplitude of the first order diffracted beam can be modulated by using the Lee method. In order to allow a DMD to modulate two beams simultaneously, a multiplexed computer hologram is loaded on the DMD, the multiplexed computer hologram is generated by superimposing two separate holograms, each of which encodes a conjugate wavefront that generates an excitation focus and a STED focus, and superimposing a linear grating phase factor on each of the two separate holograms, the linear grating phase factor adjusting the diffraction angle of the primary beam, and the frequencies of the two linear phase grating factors are set so that the primary beams diffracted by the two computer holograms overlap each other, as shown in fig. 6 (b). Thus, the primary light creates a superimposed wavefront. For clarity, in this figure, no diffraction orders other than the zeroth and first order diffraction orders are exhibited. The modulated beam is then directed through objective OBJ1(10X, NA ═ 0.25) onto a deposited ZnO scattering layer. In this example, a ZnO scattering layer is used as the strong scattering medium, and its thickness is about 280 μm. After passing through the ZnO scattering layer, the light beam was collected by an objective OBJ2(20X, NA ═ 0.4), then imaged by a convex lens L3, and the light intensity was recorded with a CMOS camera (D752, PixeLINK), as shown in fig. 6 (c). To generate the excitation and STED focus behind the scattering medium, the TM of both beams needs to be obtained first. TM is measured by phase shift method, and two plane waves are introduced as reference beams. In the present embodiment, the input mode set on the DMD is N-32 × 32, the output mode on the camera is M-480 × 480 pixels, and a high signal-to-noise ratio is obtained by encoding the input mode with the Hadamard basis vector. It should be noted that the calibration module in fig. 6(c) can be removed from the ZnO scattering medium layer after the TM is measured, and then the STED imaging can be performed.
The detailed steps are the same as those of the beam generation method for stimulated radiation loss imaging through a strong scattering medium provided in the first embodiment, and are not described herein again.
The above description is only a preferred embodiment of the present disclosure and is not intended to limit the present disclosure, and various modifications and changes may be made to the present disclosure by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present disclosure should be included in the protection scope of the present disclosure.
Claims (10)
1. A method of generating a beam for stimulated emission depletion imaging through a strongly scattering medium, comprising the steps of:
acquiring an excitation beam and a loss beam;
obtaining a combined wavefront of the shaped excitation light beam and the shaped loss light beam according to the obtained excitation light beam, the obtained loss light beam and the transmission matrix;
generating a light beam according to the shaped combined wavefront and the strong scattering medium;
the combined wave front of the shaped excitation beam and the loss beam uses a transmission matrix and a multiplexing computation holographic technology to realize the overlapping of the excitation beam and the loss beam on time and space.
2. A method of generating a beam for stimulated emission depletion imaging through a strongly scattering medium as claimed in claim 1, wherein after the acquisition of the excitation and depletion beams, a calibration of the excitation and depletion beam transmission matrices is required.
3. A method of generating a beam for stimulated emission depletion imaging through a strongly scattering medium as claimed in claim 2, wherein the excitation beam transmission matrix and the depletion beam transmission matrix are completely uncorrelated, and both transmission matrices are measured separately.
4. A method of generating a beam for stimulated emission depletion imaging through a strongly scattering medium as claimed in claim 3, wherein the measured transmission matrix of the excitation beam is optically phase-conjugated to obtain a conjugate wavefront, and the desired excitation focus is formed on the output plane based on the obtained conjugate wavefront.
5. A method of generating a beam for stimulated emission depletion imaging through a strongly scattering medium as claimed in claim 4, characterized in that the measured depletion beam is modulated with a point spread function to obtain a depletion focus in a point spread distribution.
6. The method of claim 5, wherein the spatial light modulator is used to load a multiplexed computer hologram to shape the excitation and loss beams simultaneously, and the shaped combined wavefront is incident on the strong scattering medium to create an overlapping focus to generate the beam.
7. A beam generation system for stimulated radiation depletion imaging through a strongly scattering medium, comprising:
a laser for acquiring an excitation beam and a loss beam;
the light beam processing module adjusts the focus of the excitation light beam and the focus of the loss light beam to be overlapped on time and space based on the transmission matrix and the multiplexing calculation holographic technology, and performs wave front shaping on the laser light beam and the loss light beam to obtain the combined wave front of the shaped excitation light beam and the loss light beam;
and the light beam generation module is used for transmitting the shaped combined wavefront through a strong scattering medium to generate a light beam.
8. A beam generation system for stimulated radiation depletion imaging through a strongly scattering medium as claimed in claim 7, wherein a beam splitter and a beam expander are disposed between said laser and said beam processing module in sequence; and the excitation light beam and the loss light beam both adopt continuous light sources.
9. A beam generation system for stimulated emission depletion imaging through a strongly scattering medium as claimed in claim 7, wherein said beam processing module employs digital micromirror devices including beam alignment modules, beam spatial processing units and beam temporal processing units.
10. A beam generation system for stimulated emission depletion imaging through a strongly scattering medium as claimed in claim 9, wherein said beam alignment module subjects said acquired excitation beam to an optical phase conjugation of a transmission matrix to obtain an excited focused beam, and subjects said acquired depletion beam to a point spread function modulation of said transmission matrix to obtain a depleted focused beam.
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