WO2020034299A1 - 一种并行多区域成像装置 - Google Patents

一种并行多区域成像装置 Download PDF

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Publication number
WO2020034299A1
WO2020034299A1 PCT/CN2018/106072 CN2018106072W WO2020034299A1 WO 2020034299 A1 WO2020034299 A1 WO 2020034299A1 CN 2018106072 W CN2018106072 W CN 2018106072W WO 2020034299 A1 WO2020034299 A1 WO 2020034299A1
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Prior art keywords
light
module
spatial
scanning
sample
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English (en)
French (fr)
Inventor
吕晓华
曾绍群
胡庆磊
李培
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Huazhong University of Science and Technology
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Huazhong University of Science and Technology
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Priority to US16/304,305 priority Critical patent/US11409092B2/en
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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0036—Scanning details, e.g. scanning stages
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/008—Details of detection or image processing, including general computer control
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/02—Objectives
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/56—Cameras or camera modules comprising electronic image sensors; Control thereof provided with illuminating means

Definitions

  • the invention belongs to the field of imaging, and more particularly, relates to a parallel multi-region imaging device.
  • Imaging technology is used to obtain spatial information of the sample.
  • Wide-field imaging technology uses area array detection to obtain light information in the imaging area in one exposure time. It is a way to obtain information in parallel. Although this method of acquiring information in parallel is fast (up to several thousand frames per second), it has no tomographic capability.
  • Tomography refers to acquiring only the signals in a certain thin layer (for example, the focal plane) and shielding the signals in other areas in the space (for example, out of the focal plane), thereby reducing the mutual interference of signals at different positions in the space, thereby obtaining high resolution High-contrast images.
  • the imaging technology with tomographic capabilities mainly includes confocal imaging technology and non-linear imaging technology.
  • Confocal imaging technology sets a small hole conjugated to the object point on the image side of the imaging system, and uses spot lighting to focus the illumination light on the object point.
  • the signal from the object point is imaged by the imaging system. Converge in the small holes on the image side, pass through the small holes, and then be received by the detector. Although some other areas outside the object point still have a certain intensity of illumination light, the signals from these locations cannot pass through the small holes after being passed through the imaging system, cannot be received by the detector, and are thus isolated.
  • Confocal imaging technology modulates the illumination intensity in the object space through point illumination, and then isolates the signals at other positions outside the object point through small holes to obtain the tomographic effect.
  • a scanner is required to perform the point Scan, detect point by point, and stitch the signals of each point together to form an image.
  • Non-linear imaging technology is an imaging technology based on non-linear effects. Because the intensity of the nonlinear effect has a high-order nonlinear relationship with the space-time density of the photon, it has many advantages that are different from the linear imaging technology, such as high resolution and optical tomography. Due to this non-linear feature, the traditional non-linear imaging technology needs to focus the illumination light to ensure that a sufficiently strong non-linear effect can be generated in the focal point, and then scan with a scanner to form an image.
  • the above two imaging technologies with tomographic capabilities both require scanning imaging, and scanning a surface or even a body in space is very time consuming. And the imaging speed is very important to observe the dynamic process of the sample.
  • a parallel detection mode can be introduced, such as extending a single-point scan to a multi-point simultaneous scan or even a line scan.
  • a parallel detection mode can be introduced, such as extending a single-point scan to a multi-point simultaneous scan or even a line scan.
  • a large number of methods and technologies have been put into practice, but most of them only perform parallel detection on a horizontal two-dimensional plane relative to the optical axis of the imaging system. Due to the object-image relationship, it is difficult to analyze the spatial regions of different axial positions. Perform parallel probing. Although several methods have been developed in the industry for parallel detection of spatial regions with different axial positions, they are very limited.
  • One method is to use pulsed illumination to multiplex signals at different axial positions in time, and then demodulate the acquired timing signals to perform parallel imaging.
  • the limitations of this method are as follows: First, the method requires a high-speed signal acquisition system. Because the gain bandwidth product of the signal amplifier is limited, high speed means a large bandwidth, resulting in small gain, which makes it difficult to perform weak light detection. Second, the sample pair The illumination light has a fast response. When the sample is a fluorescent sample, some fluorescent substances have a longer fluorescence lifetime, which will cause signals at different positions to be aliased in time, which makes it impossible to demodulate.
  • the second method is to collect the superimposed signals of multiple spatial regions of different axial positions without distinction, and then use the prior knowledge of the sample to perform algorithmic image processing to separate the superimposed signals into individual signals of each spatial region.
  • the limitations of this method are as follows: firstly, it needs to have sufficient prior knowledge about the sample; secondly, it needs the signals in the space to be sparse; if the signals are dense in the space, the algorithm is prone to misjudgment.
  • the present invention provides a parallel multi-region imaging device, thereby solving the difficulty of weak light detection, signal demodulation, and error easily in the existing parallel multi-region imaging detection technology. Judgment and other technical issues.
  • the present invention provides a parallel multi-region imaging device, including: a multi-focus generating module, a spatial demodulation module, and a detection module;
  • the multi-focus generating module is configured to modulate the illumination light to generate multiple focal points on the object side to form a plurality of different illumination areas, and then generate multiple signal lights through the sample to be imaged;
  • the spatial demodulation module It is used to make the energy distribution of each signal light no longer overlap or the overlapping area is smaller than the target requirement;
  • the detection module is configured to independently detect each signal light after the spatial demodulation module, so as to implement parallel multi-region imaging.
  • the device further includes an optical splitter;
  • the beam splitter is configured to spatially separate the illumination light and the signal light returned from the sample to be imaged, so that the illumination light output by the multi-focus generating module is transmitted to the sample to be imaged, and at the same time, the light from the sample to be imaged is transmitted.
  • the signal light returned by the imaging sample is propagated to the spatial demodulation module.
  • the light splitting method of the spectroscope includes separation according to wavelength, separation according to polarization state, separation according to spatial position, and attenuation separation.
  • the device further includes a scanning module
  • the scanning module is configured to change a propagation direction, a divergence degree, or a convergence degree of a light beam, so that a focal point or a one-dimensionally focused line of an objective lens changes position in space to perform scanning.
  • the scanning device in the scanning module includes a reflection type, a transmission type or a diffractive type.
  • the device further includes: a relay optical path; the relay optical path is used to achieve pupil matching between the scanning module and an objective lens so as to be within a target scanning range of the scanning module The inside allows the illumination light to enter the objective lens to illuminate the sample to be imaged.
  • the above technical solutions conceived by the present invention can achieve the following beneficial effects: using the multi-focus generation module to generate a plurality of different illumination areas on the object side, and then generating multiple signal lights, using The spatial demodulation module makes the energy distribution of the signal light in the space no longer overlap (or only overlap), and then uses the detection module to detect the signal. Therefore, the technical problems of the existing parallel multi-region imaging detection technology that are difficult to perform low-light detection, signal demodulation, and misjudgment can be solved.
  • FIG. 1 is a schematic structural diagram of a device according to an embodiment of the present invention.
  • FIG. 2 is a schematic structural diagram of another device according to an embodiment of the present invention.
  • FIG. 3 is a schematic structural diagram of a multi-focus generation module according to an embodiment of the present invention.
  • FIG. 4 is a schematic structural diagram of another multi-focus generating module according to an embodiment of the present invention.
  • FIG. 5 is a schematic structural diagram of an optical splitter provided by an embodiment of the present invention.
  • FIG. 6 is a schematic structural diagram of another optical splitter provided by an embodiment of the present invention.
  • FIG. 7 is a schematic structural diagram of another optical splitter provided by an embodiment of the present invention.
  • FIG. 8 is a schematic structural diagram of a spatial demodulation module according to an embodiment of the present invention.
  • FIG. 9 is a schematic structural diagram of another spatial demodulation module according to an embodiment of the present invention.
  • FIG. 10 is a schematic structural diagram of another spatial demodulation module according to an embodiment of the present invention.
  • FIG. 11 is a schematic structural diagram of a device according to an embodiment of the present invention.
  • FIG. 12 is a schematic structural diagram of another device according to an embodiment of the present invention.
  • FIG. 13 is a schematic structural diagram of another device according to an embodiment of the present invention.
  • FIG. 14 is a schematic structural diagram of another device according to an embodiment of the present invention.
  • 15 is a schematic structural diagram of another device according to an embodiment of the present invention.
  • FIG. 16 is a schematic structural diagram of another device according to an embodiment of the present invention.
  • the invention is a device for realizing parallel multi-region imaging using a certain structure.
  • the imaging in the present invention refers to acquiring optical information in space.
  • FIG. 1 is a schematic structural diagram of a device according to an embodiment of the present invention, including a light source module 1, a preprocessing module 2, a multi-focus generating module 3, a beam splitter 4, a scanning module 5, a relay optical path 6, and an objective lens 7. , Space demodulation module 8, detection module 9, and control acquisition processing module 10.
  • the light source module is used for generating illumination light.
  • the light source can be a non-coherent light source (such as a xenon lamp, a halogen lamp, a light emitting diode LED, etc.), or a coherent light source (such as a continuous laser, a pulse laser, a super-radiation light emitting diode SLD, etc.).
  • Monochromatic light source the type of light source including its emission spectrum is determined according to the imaging sample and imaging needs.
  • the light source module may also include other auxiliary devices, such as a collimating lens that collimates the output light or a shaper that shapes the output light spot.
  • the pre-processing module is used to process the light beam to meet the requirements of subsequent optical paths, for example, to achieve functions such as beam expansion, polarization adjustment, and power attenuation.
  • the multi-focus generation module is used to modulate the illumination light so that the object side of the final objective lens generates two or more focal points (or two or more one-dimensionally focused lines).
  • the form of the multi-focus generating module includes: forming a plurality of beams through a combination of beam splitters, and then adjusting the light propagation direction and convergence and divergence using a combination of lenses, and then combining the multiple beams to form a multi-focus beam; spatial light Light modulation devices such as modulators (Spatial Light Modulator, which performs array modulation of the intensity or phase of light) or digital micromirror devices (Digital Micromirror Device, which modulates the intensity of light) and their auxiliary devices ( (Such as a dispersion compensation device, a diaphragm that stops unwanted diffraction orders, etc.) to generate a multi-focus beam; an acousto-optic deflector or acousto-optic lens composed of acousto-optic deflectors;
  • the beam splitter is used to separate the illumination light and the signal light in space, so that they do not eventually overlap in space and are far apart, so that the illumination light output by the multi-focus generation module is transmitted to the scanning module, and the signal light returned by the self-scanning module is also transmitted.
  • the splitting method of the spectroscope includes separation according to wavelength, separation according to polarization, separation according to spatial position, and attenuation separation. Separation according to the wavelength means that when the wavelengths of the illumination light and the signal light are different, the propagation paths of the illumination light and the signal light are not overlapped by using a wavelength-sensitive device.
  • a long-pass dichroic mirror is used so that the illumination light is transmitted through the dichroic mirror and the signal light is reflected by the dichroic mirror; or a short-pass dichroic mirror is used, The illumination light is reflected by the dichroic mirror and the signal light is transmitted through the dichroic mirror.
  • Separation according to the polarization state means that when the illumination light and the signal light are both polarized light, the polarization sensing device is used so that the propagation paths of the illumination light and the signal light do not overlap.
  • a polarization beam splitter can be used, so that one of the illumination light and the signal light is transmitted through the polarization beam splitter and the other is reflected by the polarization beam splitter. If the illumination light transmitted to the sample and the signal light returned by the sample are polarized light, but there is no significant difference in polarization state, you can also adjust the polarization state of the illumination light and polarized light by inserting a polarization adjustment device in the optical path. The difference is then split using a polarization sensitive device.
  • one of the optical paths between the polarization sensitive device for spectrometry and the sample can be used.
  • a quarter-wave plate is added to the position, and the polarization state will rotate after the light passes through the quarter-wave plate two times before and after, so that the polarization states of the illumination light and the signal light at the polarization sensitive device are different and then split.
  • the position and angle of the quarter-wave plate are properly placed, the polarization state of the signal light and the illumination light at the polarization sensitive device for beam splitting will be just perpendicular.
  • a polarization beam splitter has the best beam splitting effect. Separation according to spatial position means that when the illumination light and the signal light do not actually overlap completely in space but are close to each other, it is not convenient to detect the signal light separately, using optical devices such as lenses, telescopes, reflectors, prisms to amplify the illumination light and The difference in the propagation path of the signal light makes it easier to detect the signal light. For example, when the beam directions of the illumination light and the signal light are very close and difficult to distinguish, but the spatial positions do not overlap, a reflector can be used so that it does not affect the propagation of the illumination light but reflects the signal light, so that the signal light and the illumination light are in the Clearly distinguished in space.
  • Attenuation separation refers to not using the difference in physical properties of illumination light and signal light, but directly using the method of sacrificing energy to separate illumination light and signal light in space.
  • a transflective mirror or other transmissive lens
  • the reflected part is not used and only the transmitted part is used for illumination.
  • the signal light is also transmitted partly. Part of the reflection, only the reflected signal light is detected.
  • the illumination light also has a reflection part, because the reflection direction of the illumination light and the signal light is different at this time, they can be separated in space.
  • the selected transmission reflection relationship can also be used. In turn (that is, the part reflected by the illumination light is used for illumination), the final effect can separate the propagation paths of the illumination light and the reflected light in space.
  • the scanning module is used to change the propagation direction or divergence degree (or convergence degree) of the light beam, so that the focal point (or one-dimensionally focused line) of the object side of the objective lens changes position in space to perform scanning.
  • the scanning device in the scanning module includes three types of reflection type, transmission type and diffraction type.
  • the types of scanning devices include galvanometer mirrors, resonant mirrors, rotating polygon mirrors, piezoelectric vibrating mirrors (scanning devices driven by piezoelectric ceramics that can rotate or translate mirrors), deformable mirrors, Electro-optic deflector (device that changes beam propagation direction based on electro-optic effect), electro-optic lens (device that changes beam convergence and divergence based on electro-optic effect), acousto-optic deflector (device that changes beam propagation direction based on acousto-optic effect) Acousto-optic lenses (combined by acousto-optic deflectors, devices that can change the degree of beam convergence and divergence, as well as the direction of the main ray in the beam), tunable acoustically induced refractive index lenses (based on mechanical vibration generated media Refractive index distribution to achieve phase modulation, devices that change the degree of beam convergence and divergence), variable focus lenses (devices that change
  • the scanning module uses a galvanometer mirror, a resonance mirror or a piezoelectric galvanometer to obtain better beam quality and transmission efficiency.
  • the scanning module also includes a dispersion compensation device whose function is to make the direction and convergence of the light of different wavelength components in the optical path consistent in space.
  • the relay optical path is used to achieve the pupil matching between the scanning module and the objective lens, that is, within a suitable scanning range of the scanning module, the illumination light can enter the objective lens as much as possible to illuminate the sample.
  • the ideal state of pupil matching is
  • the exit pupil of the scanning module is completely coincident with the entrance pupil of the objective lens (here, the exit pupil of the entrance pupil is relative to the propagation direction of the illumination light). If the exit pupil diameter of the scanning module (for a reflective scanner such as a galvanometer mirror, the exit pupil diameter is determined by the diameter of the illumination beam) is equivalent to the entrance pupil diameter of the objective lens, the magnification of the relay optical path is 1: 1;
  • the exit pupil diameter of the scanning module is not equal to the entrance pupil diameter of the objective lens.
  • the relay optical path should have a certain lateral magnification effect so that the exit pupil diameter of the scanning module is equivalent to the entrance pupil diameter of the objective lens.
  • the relay optical path should use a 4f optical system.
  • the so-called 4f optical system means that the relay optical path includes two lenses in front and rear (here the front and rear are relative to the direction of the illumination light propagation), and the front lens and the rear lens are positive. Lens, the optical axes of the two coincide, and the back focal plane of the front lens coincides with the front focal plane of the rear lens.
  • the exit pupil of the scanning module is located on the front focal plane of the front lens and the center coincides, and the entrance pupil of the objective lens is located on the rear focal plane of the rear lens and the center coincides. If the scanning module is so close to the objective lens that it does not need to be relayed through the lens, it can make the illumination light enter the objective lens as much as possible within a suitable scanning range of the scanning module to illuminate the sample. It can also be omitted.
  • Objective lenses are optical devices with focusing functions. Because it is close to the object to be imaged (ie, the sample), it is called an objective lens.
  • the objective lens includes lenses of different structures such as a single lens, a cemented lens, and a lens group, including lenses of different focusing methods such as a refractive lens, a reflective lens, and a diffractive lens, including curved lenses, gradient index lenses, and other surfaces. Type of lens.
  • the function of the spatial demodulation module is to make the position of the signal light emitted by the sample on the object side of the objective lens at different illumination focal points (or one-dimensionally focused illumination lines) significantly separated in space to detect them separately.
  • the spatial demodulation module is composed of a focusing lens and a perforated mirror.
  • the perforated mirror is a mirror with a through hole, preferably a flat mirror.
  • the position of a hole with an aperture mirror and an illumination focal point of the objective lens is in the position of an object image conjugate relationship.
  • the object image relationship is an imaging system composed of all optical devices in the process of signal light propagating to the small hole.
  • the imaging system includes (without consideration of the plane mirror in the optical path) a focusing lens, a relay optical path, and an objective lens in a spatial demodulation module.
  • a focusing lens In the simplest case, the imaging system includes (without consideration of the plane mirror in the optical path) a focusing lens, a relay optical path, and an objective lens in a spatial demodulation module.
  • each focal point or one-dimensionally focused illumination line
  • the signal light emitted by the sample at the other illuminating focal point position of the objective lens must not pass through the small hole smoothly, so these other positions Most of the emitted signal light is reflected by a perforated mirror, so a perforated mirror can be used to demodulate the signal light emitted by an object point that is in a conjugate relationship with the object image.
  • demodulation can be performed using a perforated mirror; when there are N channels of signal light, N-1 perforated mirrors can be used for demodulation.
  • a focusing lens and a perforated mirror are used to combine the small hole of the perforated mirror and the object point of a certain signal as the object image conjugate relationship. Just come out. After demodulation, each signal light eventually has different propagation paths in space.
  • the illumination is not the focus point illumination but the focus line illumination
  • the small hole on the reflector is changed to a slit that is in a conjugate relationship with the object image at the position of the focus line of the illumination.
  • the above-mentioned small holes and slits are in the object image conjugate relationship with the object-side illumination points or illumination lines, but the geometric dimensions need not strictly satisfy the object image conjugate relationship.
  • the space demodulation module includes passive and active.
  • the passive space demodulation module consists of passive devices (such as passive optical elements such as mirrors and lenses).
  • the active space demodulation module consists of active devices (such as digital micromirror devices, etc.) Active device).
  • the space demodulation module is composed of an active device
  • the active device is controlled by a control acquisition processing module.
  • the principle of spatial demodulation is: there are multiple channels of light, and the focus position of each channel is different. Use the device to individually select a channel to make the channel propagate in a certain direction and make others Roads propagate in other directions to ensure that the propagation paths of the roads are spatially separated from the propagation paths of other roads, so that the roads are demodulated separately, and multiple optical paths are demodulated in turn, so that all the roads are finally distinguished.
  • a device can only demodulate one channel of signals, it is necessary to use the above-mentioned cascade to sequentially demodulate.
  • the device itself has the function of demodulating multiple channels in parallel, it can also directly demodulate in parallel or mix parallel and cascaded demodulation. It is not necessary for a device to completely demodulate one or several signals, as long as it can distinguish multiple signals to a certain degree, and then cascade multiple devices to finally demodulate each signal. After demodulation, the propagation paths of the signals in the end no longer overlap in space, or the energy distribution of the signals in the space no longer overlaps (or only partially overlaps).
  • the detection module is used to independently detect each signal demodulated by the space demodulation module.
  • the detection module is mainly composed of photodetectors, including area array photodetectors (area array CCD detectors and area array CMOS detectors), line array photodetectors (line array CCD detectors and line array CMOS detectors), Photomultiplier tubes, photodiodes and other devices that can convert optical signals into electrical signals.
  • the number of detectors in the detection module meets the requirement of the number of signal channels as a principle.
  • the detection module also includes other devices useful for signal detection. For example, when the spot of the signal light propagates to the position of the detector's photosensitive surface is too large to cause signal loss, place a detector in front of the detector.
  • a combination of a focusing lens and a small hole is used to pass the signal light through the small hole and then enter the detector to suppress the interference.
  • controlling the acquisition and processing module include: controlling all active devices (mainly the scanner in the scanning module, but also the active devices in other modules) to make it work in the imaging process; Acquisition, including signal amplification, filtering, analog-to-digital conversion, etc., so that the original signal output by the detector can be processed data; processing of the collected signal, including operations such as stitching, grayscale transformation, etc., to make the collected data Display or store in the form of images or other data.
  • the imaging process of the parallel multi-region imaging method of the embodiment of the present invention is: the illumination light is emitted by the light source module 1, and after the pre-processing module 2 reaches an appropriate beam diameter, optical power or polarization state, it enters the multi-focus generation module 3, and more
  • the focus generation module 3 modulates the illumination light to generate two or more multiple sub-beams with different directions or degrees of focus, and then the illumination light continues to propagate to the beam splitter 4 and passes through the beam splitter 4 to the scanning module 5 and then passes through
  • the light path 6 propagates to the objective lens 7, two or more focal points (or one-dimensionally focused illumination lines) are formed on the object side by the objective lens 7; after the illumination light is irradiated on the sample 0, it is reflected, scattered, or fluorescent on the sample 0
  • the signal light is generated by the equal effect; the signal light is collected by the objective lens 7 and is transmitted to the scanning module 5 through the relay optical path 6.
  • the scanning module 5 is scanned under the control of the control acquisition processing module 10, and the signal light is transmitted to the beam splitter through the scanning module 5.
  • Device 4 the beam splitter causes the signal light to continue to propagate along a space path different from the illumination light, and then the signal light enters the space demodulation module 8, and the space demodulation module 8 separates each channel Each signal light beam generated on the sample is spatially separated.
  • the detection module 9 detects each separated signal light, and controls the acquisition processing module 10 to collect and process the electrical signals output by the detection module 9 and finally use an image. Or other data forms.
  • the embodiment of the present invention also includes other forms of structures and imaging processes, such as using two objective lenses to perform illumination and signal light collection, respectively.
  • the structural change is that the number of the scanning module, the relay optical path and the objective lens are two, and the beam splitter is placed behind the second scanning module.
  • the imaging process is: the illumination light is emitted by the light source module 1, and after the pre-processing module 2 reaches the appropriate beam diameter, optical power or polarization state, it enters the multi-focus generation module 3, and the multi-focus generation module 3 modulates the illumination light to Two or more sub-beams with different directions or degrees of focus are generated, and then the illumination light continues to propagate to the first scanning module 5a, and then propagates to the first objective lens 7a through the first relay optical path 6a.
  • the square forms two or more focal points (or one-dimensionally focused illumination lines); after the illumination light is irradiated on the sample 0, the signal light is generated on the sample 0 through reflection, scattering, or fluorescence effects; the signal light is used by the second objective lens 7b Collected and transmitted to the second scanning module 5b through the second relay light path 6b, the second scanning module 5b is scanned under the control of the control acquisition processing module 10, and the illumination light is transmitted to the spectroscope 4 through the second scanning module 5b.
  • the spectroscope 4 filters out a part of the illumination light that is mixed with the signal light, and then the signal light is propagated to the spatial demodulation module 8, and the spatial demodulation module 8 separates each of the illumination sub-beams generated on the sample.
  • the detection module 9 brightest separated signals optical detection, acquisition and processing module 10 controls the detection module 9 outputs the electrical signal collected and processed and ultimately displayed or stored image data or other forms.
  • the spatial position of the signal light after it passes through the scanning module does not change with the scanning of the scanning module, which is called descanning.
  • the scanning process of the second scanning module 5b and the first scanning module 5a should be synchronized to achieve the effect of push scanning; the function of the beam splitter 4 has also changed, and it is only necessary to alias the signal light together.
  • the illumination light can be filtered.
  • FIG. 3 A specific implementation manner 1 of a multi-focus generating module according to an embodiment of the present invention is shown in FIG. 3.
  • the half-wave plate 31a is placed in front of the polarization beam splitter prism 32a.
  • the polarization beam splitter prism 32a divides the input light into two beams of reflection and transmission.
  • the half-wave plate 31a is used to adjust the energy ratio of the reflected light and the transmitted light of the polarization beam splitter prism 32a.
  • the half-wave plate 31b is placed in front of the polarization beam splitting prism 32b.
  • the polarization beam splitting prism 32b divides the reflected light of the polarization beam splitting prism 32a into two beams.
  • the half-wave plate 31b is used to adjust the energy ratio of the reflected light and the transmitted light of the polarization beam splitting prism 32b. Utilizing the beam splitting effect of the polarization beam splitting prisms 32a and 32b, three sub-beams are finally obtained.
  • the first sub-beam is not modulated by the lens
  • the second sub-beam is modulated by the lens 34a and the lens 34b
  • the third sub-beam is modulated by the lens 34c and the lens 34d.
  • the direction and focus divergence of the modulated beam change.
  • the first sub-beam is reflected by the reflecting mirror 33a
  • the third sub-beam is reflected by the reflecting mirror 33b.
  • the first sub-beam is transmitted through the polarization beam splitting prism 32c, the second sub-beam is reflected by the polarization beam splitting prism 32c, the first and second sub-beams are combined, and then the polarization state is modulated by the half-wave plate 31c.
  • the function of the half-wave plate 31c is to make the light beams after the first and second sub-beams are combined to have a proper polarization state so that they are reflected by the polarization beam splitting prism 32d.
  • the third sub-beam is transmitted through the polarization beam splitting prism 32d. After passing through the polarization beam splitting prism 32d, the three sub-beams are all combined. At this time, although the directions and divergent focusing degrees of the three sub-beams are different, they still overlap in space.
  • FIG. 4 A specific implementation manner 2 of the multi-focus generating module according to an embodiment of the present invention is shown in FIG. 4.
  • the spatial light modulator 35 generates a specific modulation pattern under the control of the control acquisition processing module 10.
  • Different modulation patterns can generate different multifocal distributions. Specifically, the phase modulation distribution that varies linearly with space is used to change the direction of the outgoing light, and the phase modulation pattern that is distributed with space as a lens modulation function is used to change the output light. Focus divergence (the lens modulation function is circularly symmetric, and its center position can also be used to change the direction of the outgoing light).
  • the dichroic sheet 41 has different reflectances and transmittances for light of different wavelengths, and is expressed as long-wavelength light having high transmittance and short-wavelength light having high reflectivity, so that light propagation paths of different wavelengths are different.
  • a specific implementation manner 2 of the spectroscope according to the embodiment of the present invention is shown in FIG. 6.
  • the polarization beam splitter prism 42 has different reflectances and transmittances for light of different polarization states, and is expressed as being highly transparent to light with a polarization state of p and highly reflective to light with a polarization state of s, thereby allowing light of different polarization states to propagate.
  • the paths are different.
  • a specific implementation manner 3 of the spectroscope according to the embodiment of the present invention is shown in FIG. 7.
  • the edge of the reflector 43 is placed between the two beams, so that it reflects one path of light, while the other path continues to be unobstructed, so that the light in the space whose propagation paths do not overlap is further separated in the space.
  • FIG. 8 A specific implementation manner 1 of a space demodulation module according to an embodiment of the present invention is shown in FIG. 8.
  • the focal point of one path overlaps with the small hole of the perforated mirror 81, so that this path will continue to propagate through the small hole, and the focal point of the other path does not overlap with the small hole of the perforated mirror 81, so Most of this energy will be reflected by the mirror.
  • the light beam 80a that has finally passed through the pinhole and the reflected light beam 80b no longer overlap in space.
  • FIG. 9 A specific implementation manner 2 of the space demodulation module according to an embodiment of the present invention is shown in FIG. 9.
  • the digital micromirror device 82 generates a specific modulation pattern under the control of the control acquisition processing module 10. There are two paths of light in the space, one of which is focused on the working surface of the digital micromirror device 82.
  • the micromirror at the focal position of the digital micromirror device 82 is deflected to one state and the other micromirrors are deflected to another state.
  • This path of light focused on the digital micromirror device 82 is reflected by the digital micromirror device 82 to a certain direction, and the other path of light is reflected to another direction, and finally forms light beams 80c and 80d which do not overlap in space.
  • FIG. 10 A specific implementation manner 3 of the space demodulation module according to the embodiment of the present invention is shown in FIG. 10.
  • the spatial light modulator 83 generates a specific modulation pattern under the control of the control acquisition processing module 10. There are three paths of light in space, and they are focused differently.
  • the pattern on the spatial light modulator 83 is a deformed grating (off-axis Fresnel zone plate or off-axis holographic lens). It cooperates with the lens 831 to diffract three-way light with different degrees of focus to different diffraction orders. In the end, light beams 80e, 80f, and 80g that do not overlap in space are finally formed.
  • FIG. 11 A specific implementation manner 1 of the overall structure of the embodiment of the present invention is shown in FIG. 11.
  • the laser light emitted by the femtosecond laser 11 is expanded by a pre-processing module composed of a lens 21a and a lens 21b, and then is incident on a spatial light modulator 35.
  • the spatial light modulator modulates the wavefront to generate two focal points, and the multi-focus beam passes through the lens.
  • 351a and lens 351b pass through the dichroic mirror 41, and then incident on a dual-axis scanning module composed of a galvanometer mirror 51a and a galvanometer mirror 51b.
  • the relay lens 61a After scanning by the scanning module, they pass through the relay lens 61a and The relay lens 61 b is incident on the objective lens 71 and is focused on the sample 0 through the objective lens 71.
  • the femtosecond laser generates fluorescence in the sample.
  • the fluorescence signal is collected by the objective lens 71 and returned along the path.
  • it hits the dichroic mirror 41 and is reflected by the dichroic mirror 41. After the reflected light is focused by the lens 811, all the light passes through.
  • the small hole of the perforated mirror 81 is detected by the photomultiplier tube 91a, and the other path is reflected by the perforated mirror 81 and detected by the photomultiplier tube 91b.
  • the control acquisition processing module is not shown in the figure.
  • the spatial light modulator 35 and the galvanometer mirrors 51a and 51b are all controlled by the control acquisition processing module.
  • the output signals of the photomultiplier tubes 91a and 91b are collected by
  • FIG. 12 A specific implementation manner 2 of the overall structure of the embodiment of the present invention is shown in FIG. 12.
  • the laser light emitted by the femtosecond laser 11 is expanded by a pre-processing module composed of a lens 21a and a lens 21b, and then the polarization state is adjusted by the half-wave plate 31a and incident on the polarization beam splitting prism 321a.
  • the polarization beam splitting prism 321a divides the light into two paths. One passes through the lens 34a and the lens 34b to adjust the degree of convergence and divergence and is incident on the polarization beam splitting prism 321b. The other path is reflected by the reflection mirrors 331a and 331b and incident on the polarization beam splitting prism 321b.
  • the dichroic prism 321b combines two channels of light.
  • the combined light is reflected by the mirror 331c, passes through the relay of the lens 351a and the lens 351b, and enters the dichroic mirror 41 and enters a dual-axis scanning module composed of a galvanometer mirror 51a and a galvanometer mirror 51b.
  • After scanning by the scanning module it is incident on the objective lens 71 through the relay lens 61a and the relay lens 61b, and is focused on the sample 0 through the objective lens 71.
  • the femtosecond laser generates fluorescence in the sample.
  • the fluorescence signal is collected by the objective lens 71 and returned along the path. Finally, it hits the dichroic mirror 41 and is reflected by the dichroic mirror 41.
  • the control acquisition processing module is not shown in the figure.
  • the galvanometer mirrors 51a and 51b are both controlled by the control acquisition processing module.
  • the output signals of the photomultiplier tubes 91a and 91b are collected by the control acquisition processing module.
  • FIG. 13 A specific implementation manner 3 of the overall structure of the embodiment of the present invention is shown in FIG. 13.
  • the laser light emitted by the femtosecond laser 11 is expanded by a pre-processing module composed of a lens 21a and a lens 21b, and then incident on a reflection grating 361.
  • the reflection grating 361 is used to compensate the spatial dispersion of the digital micromirror device 36.
  • the emitted light is incident on the digital micromirror device 36 after passing through the dichroic mirror 41, and the digital micromirror device 36 simultaneously realizes the functions of multifocal generation and scanning.
  • the light After the light passes through the digital micromirror device 36, it is incident on the objective lens 71 through the relay lenses 61a and 61b, and the objective lens 71 focuses the light onto the focal point.
  • the femtosecond laser generates fluorescence in the sample.
  • the fluorescence signal is collected by the objective lens 71 and returned along the path.
  • the dichroic mirror 41 hits the dichroic mirror 41 and is reflected by the dichroic mirror 41.
  • the reflected light is compensated by the dispersion compensation module 49, and then processed by the dispersion compensation module 49.
  • the control acquisition processing module is not shown in the figure.
  • the digital micromirror device 36 is controlled by the control acquisition processing module.
  • the output signals of the photomultiplier tubes 91a and 91b are collected by the control acquisition processing module.
  • FIG. 14 A specific implementation manner 4 of the overall structure of the embodiment of the present invention is shown in FIG. 14.
  • the laser light emitted by the femtosecond laser 11 is expanded by a pre-processing module composed of a lens 21a and a lens 21b, and then incident on a transmission grating 352.
  • the transmission grating 352 is used to compensate the spatial dispersion of the spatial light modulator 35.
  • the emitted light is incident on the spatial light modulator 35.
  • the spatial light modulator modulates the wavefront to generate two focal points.
  • the multi-focus beam passes through the lens 351a and the lens 351b, passes through the dichroic mirror 41, and then enters the acoustic and optical
  • the femtosecond laser generates fluorescence in the sample.
  • the fluorescence signal is collected by the objective lens 71 and returned along the path. Finally, it hits the dichroic mirror 41 and is reflected by the dichroic mirror 41.
  • the reflected light is compensated by the dispersion compensation module 49, and then processed by the dispersion compensation module 49.
  • the lens 811 After the lens 811 is focused, one path of light is detected by the photomultiplier tube 91a through the small hole of the perforated mirror 81, and the other path is reflected by the perforated mirror 81 and detected by the photomultiplier tube 91b.
  • the control acquisition processing module is not shown in the figure.
  • the spatial light modulator 35 and the acousto-optic deflectors 52a and 52b are all controlled by the control acquisition processing module.
  • the output signals of the photomultiplier tubes 91a and 91b are collected by the control acquisition processing module.
  • FIG. 15 A specific implementation manner 5 of the overall structure of the embodiment of the present invention is shown in FIG. 15.
  • the laser light emitted by the femtosecond laser 11 is expanded by a pre-processing module composed of a lens 21a and a lens 21b, and then incident on a transmission grating 352.
  • the transmission grating 352 is used to compensate the spatial dispersion of the spatial light modulator 35.
  • the emitted light is incident on the spatial light modulator 35.
  • the spatial light modulator modulates the wavefront to generate two focal points.
  • the multi-focus light beam passes through the lens 351a and the lens 351b, passes through the dichroic mirror 41, and then is incident on the current detector.
  • the dual-axis scanning module composed of the counter 51a and the galvanometer mirror 51b
  • the scanning module After being scanned by the scanning module, it is incident on the objective lens 71 through the relay lens 61a and the relay lens 61b, and is focused on the sample 0 through the objective lens 71.
  • the femtosecond laser generates fluorescence in the sample.
  • the fluorescence signal is collected by the objective lens 71 and returned along the path.
  • it hits the dichroic mirror 41 and is reflected by the dichroic mirror 41.
  • the reflected light is incident on the digital micromirror device 83.
  • a deformed grating (off-axis Fresnel zone plate) is generated on the digital micromirror device 83, so that one light is focused on the small hole 381a and then detected by the photomultiplier tube 91a, and the other is focused on the small hole 831b and then photoelectrically Detection by the doubling tube 91b.
  • the control acquisition processing module is not shown in the figure.
  • the spatial light modulator 35, galvanometer mirrors 51a and 51b, and digital micromirror device 83 are all controlled by the control acquisition processing module.
  • the output signals of photomultiplier tubes 91a and 91b are controlled. Acquisition processing module acquisition.
  • FIG. 16 A specific implementation manner 6 of the overall structure of the embodiment of the present invention is shown in FIG. 16.
  • the laser light emitted by the femtosecond laser 11 is expanded by a pre-processing module composed of a lens 21a and a lens 21b, and then incident on a transmission grating 352.
  • the transmission grating 352 is used to compensate the spatial dispersion of the spatial light modulator 35.
  • the emitted light is incident on the spatial light modulator 35.
  • the spatial light modulator modulates the wavefront to generate multiple focal points.
  • the multi-focus light beam passes through the lens 351a and the lens 351b, passes through the dichroic mirror 41, and then is incident on the current detector
  • the femtosecond laser generates fluorescence in the sample.
  • the fluorescence signal is collected by the objective lens 71 and returned along the path.
  • it hits the dichroic mirror 41 and is reflected by the dichroic mirror 41.
  • the reflected light is relayed by the lenses 841a and 841b.
  • the modulator 84 is a passive modulation device, and is made of a transparent material.
  • the modulator 84 may also be a spatial light modulator or a digital micromirror device.
  • the focal depth of the focusing spot behind the lens 84 is greater than the focal depth of the lens focus at the same numerical aperture, eliminating the phenomenon of defocusing and focusing spots overlapping, and different lateral positions after focusing at different focal points. Different focal points are focused on the same plane and their lateral positions are different, that is, spatial demodulation is achieved.
  • the detector 92 is located in a focusing area behind the modulator 84. At this time, the signal light appears as a focused spot with a different lateral position on the detector 92.
  • the detector 92 is an array detector, such as a multi-anode photomultiplier tube, an array avalanche diode, an area array CMOS sensor, and an area array CCD sensor, so that signals of different focused light spots can be collected.
  • the control acquisition processing module is not shown in the figure.
  • the spatial light modulator 35, galvanometer mirrors 51a and 51b, and other active devices are all controlled by the control acquisition processing module.
  • the output signal of the detector 92 is collected by the control acquisition processing module. .

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Abstract

一种并行多区域成像装置，应用于成像领域，包括：多焦点生成模块（3）、空间解调模块（8）及探测模块（9）；多焦点生成模块（3），用于对照明光进行调制，以在物方产生多个焦点，形成多个不同的照明区域，进而通过待成像样品产生多路信号光；空间解调模块（8），用于使各路信号光在空间上的能量分布不再重叠或重叠区域小于目标要求；探测模块（9），用于对经空间解调模块（8）后的各路信号光进行独立探测，以实现并行多区域成像，可以解决现有并行多区域成像探测技术存在的难以进行弱光探测、信号无法解调及容易出现误判等的技术问题。

Description

一种并行多区域成像装置 [技术领域]
本发明属于成像领域，更具体地，涉及一种并行多区域成像装置。
[背景技术]
成像技术用于获取样品的空间信息。宽场成像技术使用面阵式探测，可在一个曝光时间获取成像区域内的光信息，是一种并行获取信息的方式。这种并行获取信息的方式虽然很快(每秒帧数可达几千)，但是没有层析能力。层析是指仅获取某个薄层(例如焦面)内的信号，屏蔽掉空间中其他区域(例如焦面外)的信号，从而减少空间中不同位置的信号的相互干扰，从而获得高分辨率高对比度的图像。具有层析能力的成像技术主要包括共聚焦成像技术和非线性成像技术。
共聚焦成像技术在成像系统的像方设置一个与物方的物点共轭的小孔，成像时采用点照明方式将照明光汇聚于该物点，该物点发出的信号经成像系统成像后汇聚于像方的小孔并通过小孔，然后被探测器所接收。虽然物点外的部分其他区域仍有一定强度的照明光，但是这些位置发出的信号经成像系统后无法通过小孔，不能被探测器接收，从而被隔离掉。共聚焦成像技术通过点照明对物方空间中的照明强度进行调制，再通过小孔隔离掉物点外其他位置的信号，从而获得层析效果。由于照明和探测均只针对空间中的某个点(并非无限小点，而是相对于成像范围来说一个很小的三维区域)，为获得样品在空间上的信息，需要使用扫描器进行点扫描，逐点探测，再将每个点的信号拼接在一起形成图像。
非线性成像技术是一种基于非线性效应的成像技术。由于非线性效应的强度与光子的时空密度具有高阶的非线性关系，因此具有许多区别于线性成像技术的优点，例如分辨率高、具有光学层析能力等。也由于该非线性特征，传统的非线性成像技术需要将照明光进行聚焦，以确保焦点内能够 产生足够强的非线性效应，然后使用扫描器进行扫描以形成图像。以上两种具有层析能力的成像技术均需要扫描成像，而空间中对一个面甚至体进行扫描非常耗时。而成像速度对于观测样品的动态过程至关重要。即便使用高速的扫描器，由于对空间中的点采用串行模式进行曝光，单点曝光时间也非常有限，对信号强度和信噪比不利。为提高成像速度，可引入并行探测的模式，例如将单点扫描扩展为多点同时扫描甚至线扫描。目前已经有大量方法和技术付诸实践，但是大多数只是在相对于成像系统光轴来说一个横向的二维平面上进行并行探测，由于物像关系的原因难以对不同轴向位置的空间区域进行并行探测。虽然业内已经发展出来几种对不同轴向位置的空间区域进行并行探测的方法，但是局限性很大。方法之一是采用脉冲式照明，使不同轴向位置的信号在时间上复用，然后将采集到的时序信号进行解调从而进行并行成像。该方法的局限性表现为：首先，该方法需要高速的信号采集系统，由于信号放大器的增益带宽积有限，速度高意味着带宽大，导致增益小，难以进行弱光探测；其次，需要样品对照明光有快速的响应，当样品为荧光样品时，有的荧光物质具有较长的荧光寿命，会导致不同位置的信号在时间上混叠，从而无法解调。方法之二是不加区分的采集到不同轴向位置的多个空间区域的叠加信号后，利用对样品的先验知识采用算法进行图像处理从而将叠加的信号分离为各个空间区域单独的信号。该方法的局限性表现为：首先，需要具有足够的关于样品的先验知识；其次，需要空间中的信号较为稀疏，如果空间中信号较为密集该算法容易出现误判。
[发明内容]
针对现有技术的以上缺陷或改进需求，本发明提供了一种并行多区域成像装置，由此解决现有并行多区域成像探测技术存在的难以进行弱光探测、信号无法解调及容易出现误判等的技术问题。
为实现上述目的，本发明提供了一种并行多区域成像装置，包括：多 焦点生成模块、空间解调模块及探测模块；
所述多焦点生成模块，用于对照明光进行调制，以在物方产生多个焦点，形成多个不同的照明区域，进而通过待成像样品产生多路信号光；所述空间解调模块，用于使各路信号光在空间上的能量分布不再重叠或重叠区域小于目标要求；
所述探测模块，用于对经所述空间解调模块后的各路信号光进行独立探测，以实现并行多区域成像。
优选地，在所述多焦点生成模块和所述空间解调模块之间，所述装置还包括分光器；
所述分光器，用于将照明光和由待成像样品返回的信号光在空间上分离，以使所述多焦点生成模块输出的照明光传播至所述待成像样品，同时使自所述待成像样品返回的信号光传播至所述空间解调模块。
优选地，所述分光器的分光方法包括按照波长分离、按照偏振态分离、按照空间位置分离及衰减式分离。
优选地，在所述分光器之后，所述装置还包括扫描模块；
所述扫描模块，用于改变光束的传播方向、发散程度或汇聚程度，使得物镜物方的焦点或一维聚焦的线在空间中改变位置，从而进行扫描。
优选地，所述扫描模块中的扫描器件包括反射式、透射式或衍射式。
优选地，在所述扫描模块之后，所述装置还包括：中继光路；所述中继光路，用于实现所述扫描模块与物镜的光瞳匹配，以在所述扫描模块的目标扫描范围内使得照明光均能进入到所述物镜中从而照明到所述待成像样品上。
总体而言，通过本发明所构思的以上技术方案与现有技术相比，能够取得下列有益效果：利用多焦点生成模块在物方产生多个不同的照明区域，进而产生多路信号光，利用空间解调模块使各路信号光最终在空间上的能量分布不再重叠(或仅有极少重叠)，再利用探测模块对信号进行探测。从 而可以解决现有并行多区域成像探测技术存在的难以进行弱光探测、信号无法解调及容易出现误判等的技术问题。
[附图说明]
图1是本发明实施例提供的一种装置的结构示意图；
图2是本发明实施例提供的另一种装置的结构示意图；
图3是本发明实施例提供的一种多焦点生成模块的结构示意图；
图4是本发明实施例提供的另一种多焦点生成模块的结构示意图；
图5是本发明实施例提供的一种分光器的结构示意图；
图6是本发明实施例提供的另一种分光器的结构示意图；
图7是本发明实施例提供的另一种分光器的结构示意图；
图8是本发明实施例提供的一种空间解调模块的结构示意图；
图9是本发明实施例提供的另一种空间解调模块的结构示意图；
图10是本发明实施例提供的另一种空间解调模块的结构示意图；
图11是本发明实施例提供的一种装置的结构示意图；
图12是本发明实施例提供的另一种装置的结构示意图；
图13是本发明实施例提供的另一种装置的结构示意图；
图14是本发明实施例提供的另一种装置的结构示意图；
图15是本发明实施例提供的另一种装置的结构示意图；
图16是本发明实施例提供的另一种装置的结构示意图。
[具体实施方式]
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合附图及实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。此外，下面所描述的本发明各个实施方式中所涉及到的技术特征只要彼此之间未构成冲突就可以相互组合。
本发明为利用一定结构实现并行多区域成像的装置。本发明所述成像 是指获取空间中的光信息。
如图1所示为本发明实施例提供的一种装置的结构示意图，包括光源模块1、预处理模块2、多焦点生成模块3、分光器4、扫描模块5、中继光路6、物镜7、空间解调模块8、探测模块9及控制采集处理模块10。
光源模块用于产生照明光。光源可为非相干光源(例如氙灯、卤素灯、发光二极管LED等)，也可为相干光源(例如连续激光器、脉冲激光器、超辐射发光二极管SLD等)，可为宽带光源，也可为窄带或单色光源，光源的种类包括其发射光谱根据成像的样品和成像需求来确定。光源模块除了光源本身，还可以包括其他辅助器件，如将输出光准直的准直透镜或对输出光斑进行整形的整形器。
预处理模块用于对光束进行处理，使其满足后续光路的要求，例如实现扩束、偏振态调节、功率衰减等功能。
多焦点生成模块用于对照明光进行调制，以使最终物镜的物方产生两个或多个焦点(或两个或多个一维聚焦的线)。多焦点生成模块的形式包括：通过分光镜的组合构成多个光束，再使用透镜的组合进行光传播方向和汇聚发散程度的调整，然后再将多个光束合束构成多焦点光束；由空间光调制器(Spatial Light Modulator，对光的强度或相位进行阵列式调制的器件)或数字微镜器件(Digital Micromirror Device，对光的强度进行数字式调制的器件)等光调制器件及其辅助器件(例如色散补偿器件、拦掉多余衍射级次的光阑等)生成多焦点光束；声光偏转器或声光偏转器组成的声光透镜；由透明材料加工而成的具有相位调制作用的相位板，如衍射光栅、菲涅尔透镜、多焦点相位板等；强度型的无源衍射器件，如达曼光栅、菲涅尔波带片等；无源的强度和相位混合型的衍射器件；透镜阵列；其他形式的多焦点生成方法。当多焦点生成模块包含主动器件时(例如上述的空间光调制器、数字微镜器件、声光偏转器等)，多焦点生成模块由控制采集处理模块进行控制。
分光器用于将照明光和信号光在空间上分离，使其最终不在空间上重叠并相距较远，使多焦点生成模块输出的照明光传播至扫描模块，也使自扫描模块返回的信号光传播至空间解调模块。分光器的分光方法包括按照波长分离、按照偏振态分离、按照空间位置分离、衰减式分离等。按照波长分离是指当照明光和信号光波长有差异时，利用波长敏感器件使照明光和信号光的传播路径不重叠。例如当照明光的波长大于信号光波长时，使用长通的二向色镜，使得照明光经二向色镜透射而信号光被二向色镜反射；或使用短通的二向色镜，使得照明光被二向色镜反射而信号光经二向色镜透射。按照偏振态分离是指当照明光和信号光均为偏振光时，利用偏振敏感器件使照明光和信号光的传播路径不重叠。例如当照明光和信号光偏振态垂直时，可使用偏振分光镜，让照明光和信号光当中的一个经偏振分光镜透射而另一个被偏振分光镜反射。如果传播到样品的照明光和样品返回的信号光虽然为偏振光，但是没有偏振态上的显著差异，也可以通过在光路中插入偏振调节器件来调节照明光和偏振光的偏振态使其出现差异，然后使用偏振敏感器件进行分光。例如，当光源发出的光为线偏振光，而样品本身返回的信号光与照射到样品上的照明光同一偏振态时，可在分光用的偏振敏感器件和样品之间的光路中的某个位置加入四分之一波片，光前后两次通过四分之一波片后偏振态会发生旋转，从而使得位于偏振敏感器件处的照明光和信号光偏振态不同进而分光。当四分之一波片的位置和角度放置合适时，位于分光用偏振敏感器件处信号光和照明光的偏振态会刚好垂直，此时使用偏振分光镜有最佳的分光效果。按照空间位置分离是指当照明光和信号光在空间上其实不完全重叠但相隔较近，不便于单独对信号光进行探测时，利用透镜、望远镜、反射镜、棱镜等光学器件放大照明光和信号光在传播路径上的差异，从而方便对信号光进行探测。例如，当照明光和信号光的光束方向非常接近难以区分但是空间位置并不重叠时，可以利用一个反射镜，使其不影响照明光的传播但是反射信号光，从而将 信号光和照明光在空间上明显区分开。衰减式分离是指不利用照明光和信号光物理属性上的差异，而直接使用牺牲能量的方法使照明光和信号光在空间上分离。例如，使用一个半透半反镜(或其他透射反射比的镜片)，使得照明光透射一部分反射一部分，对于反射的那部分不加以利用而仅用透射的那部分进行照明，信号光也透射一部分反射一部分，仅探测反射的那部分信号光，虽然照明光也有反射的部分，但是由于此时照明光和信号光的反射方向不同，因此可以在空间上分离，当然对于选用的透射反射关系也可以反过来(即使用照明光反射的那一部分进行照明)，最终效果能使照明光和反射光的传播路径在空间上分离即可。
扫描模块用于改变光束的传播方向或发散程度(或汇聚程度)，使得物镜的物方的焦点(或一维聚焦的线)在空间中改变位置，从而进行扫描。扫描模块中的扫描器件包括反射式、透射式、衍射式三种。具体来讲，扫描器件的类型包括：检流计镜、共振镜、旋转多面镜、压电振镜(由压电陶瓷驱动的可使反射镜转动或平移的扫描器件)、可变形反射镜、电光偏转器(基于电光效应的改变光束传播方向的器件)、电光透镜(基于电光效应的改变光束会聚和发散程度的器件)、声光偏转器(基于声光效应的改变光束传播方向的器件)、声光透镜(由声光偏转器组合而成，可改变光束会聚和发散程度，亦可改变光束中的主光线方向的器件)、可调谐声致梯度折射率透镜(基于机械振动产生介质中折射率分布从而实现相位调制，改变光束汇聚和发散程度的器件)、可变焦透镜(基于机械形变或电浸润效应，通过改变介质形状从而改变光束会聚和发散程度的器件)、空间光调制器(Spatial Light Modulator，对光的强度或相位进行阵列式调制的器件)、数字微镜器件(Digital Micromirror Device，对光的强度进行数字式调制的器件)等。优选的，扫描模块使用检流计镜、共振镜或压电振镜，以获得更佳的光束质量和传输效率。当使用存在色散的扫描器件(例如声光偏转器)时，扫描模块还包括色散补偿器件，其功能是使光路中不同波长成分的光 在空间中的方向和汇聚发散程度一致。
中继光路用于实现扫描模块与物镜的光瞳匹配，即在扫描模块的一个合适的扫描范围内使得照明光均能尽可能进入到物镜中从而照明到样品上，光瞳匹配的理想状态是扫描模块的出瞳与物镜的入瞳完全重合(此处的入瞳出瞳是相对于照明光的传播方向来讲)。如果扫描模块的出瞳直径(对于反射式扫描器例如检流计镜，出瞳直径由照明光束的直径决定)与物镜的入瞳直径相当，则中继光路的放大率为1:1；如果扫描模块的出瞳直径与物镜的入瞳直径不相当，优选的，中继光路应有一定横向放大效果使得扫描模块的出瞳直径与物镜的入瞳直径相当。优选的，中继光路应使用4f光学系统，所谓4f光学系统是指中继光路包含前后两个透镜(此处前后是相对于照明光的传播方向来讲)，前透镜和后透镜均为正透镜，二者光轴重合，且前透镜的后焦面与后透镜的前焦面重合。优选的，扫描模块的出瞳位于前透镜的前焦面上且中心重合，物镜的入瞳位于后透镜的后焦面上且中心重合。如果扫描模块与物镜距离很近以至于不需要通过透镜进行中继即可以使得在扫描模块的一个合适的扫描范围内使得照明光均能尽可能进入到物镜中从而照明到样品上，中继光路也可以省略。
物镜是具有聚焦功能的光学器件。因为其靠近待成像物体(即样品)所以称为物镜。具体来讲，物镜包括单透镜、胶合透镜、透镜组等不同结构的透镜，包括折射型透镜、反射型透镜、衍射型透镜等不同聚焦方式的透镜，包括曲面透镜、梯度折射率透镜等不同面型的透镜。
空间解调模块的功能是使物镜物方由不同照明焦点(或一维聚焦的照明线)位置上样品发出的信号光在空间上的位置显著分离，从而分别对其进行探测。优选的，空间解调模块由聚焦透镜和带孔反射镜构成。带孔反射镜为带有通孔的反射镜，优选为平面反射镜。带孔反射镜的孔与物镜物方的某个照明焦点在位置上为物像共轭关系，该物像关系由信号光传播至该小孔的过程中的所有光学器件组成的成像系统构成，在最简单的情况下 该成像系统包括(不考虑光路中的平面反射镜)空间解调模块中的聚焦透镜、中继光路和物镜。物镜物方存在多个照明焦点(或一维聚焦的照明线)，每一个焦点(或一维聚焦的照明线)均会产生一路信号光。信号光在进入空间解调模块之前，几路信号光在空间上是存在重叠的，难以将它们区分。由于带孔反射镜的孔与物镜物方的某个照明焦点为物像共轭关系，那么物镜物方的其他照明焦点位置上由样品发出的信号光必然无法顺利通过小孔，于是这些其他位置发出的信号光绝大部分被带孔反射镜所反射，于是利用一个带孔反射镜就可以将与该小孔为物像共轭关系的物点发出的信号光解调出来。当信号光只有两路时，使用一个带孔反射镜即可进行解调；当信号光有N路时，使用N-1个带孔反射镜即可进行解调。当信号光有多路时，使用聚焦透镜和带孔反射镜进行组合，利用带孔反射镜的小孔与某路信号的物点为物像共轭关系的特点，将各路信号依次解调出来即可。解调后，各路信号光最终在空间上有不同的传播路径。当照明不是聚焦点照明而是聚焦线照明时，把反射镜上的小孔改成与照明的聚焦线位置上为物像共轭关系的狭缝。上述小孔和狭缝与物方的照明点或照明线在位置上为物像共轭关系，但是几何尺寸上不需要严格满足物像共轭关系。带孔(或狭缝)反射镜并不能保证某路信号百分百通过小孔(或狭缝)，也不能保证其他路信号百分百不通过小孔(或狭缝)而被反射，但是要保证相应路的信号过小孔(或狭缝)部分的信号强度大于被反射部分的信号强度，而其他各路的信号被反射部分的信号强度大于过小孔(或狭缝)部分的信号强度。空间解调模块包括被动式和主动式，被动式的空间解调模块由被动器件组成(例如反射镜、透镜等被动光学元件)，主动式的空间解调模块由主动器件组成(例如数字微镜器件等主动器件)。当空间解调模块由主动器件组成时，主动器件由控制采集处理模块进行控制。总而言之，无论具体采用哪种器件，其空间解调原理均为：存在多路光，每一路的聚焦位置都不同，利用器件将某一路单独选出，使该路往某个方向传播，使其他路往别 的方向传播，保证该路与其他路的传播路径在空间上分离，从而单独解调出该路，对多路光依次解调，从而最终将所有路都区分开。当然，如果一个器件只能解调出一路信号，必然要使用上述级联的方式依次解调。如果器件本身具有并行解调多路的功能，也可以直接并行解调或者并行与级联混合解调。一个器件也并非一定要完全将一路或几路信号解调出来，只要能对多路信号进行一定程度上的区分，然后采用多个器件级联，最终能把各路信号解调出来即可。解调后，最终各路信号的传播路径在空间上不再重叠，或者说各路信号在空间中的能量分布不再重叠(或仅有极少部分重叠)。
探测模块用于将空间解调模块解调后的各路信号进行独立的探测。探测模块主要由光电探测器组成，包括面阵式光电探测器(面阵CCD探测器和面阵CMOS探测器)、线阵式光电探测器(线阵CCD探测器和线阵CMOS探测器)、光电倍增管、光电二极管等能够将光信号转换为电信号的器件。探测模块中的探测器数量以满足信号通道数量要求为原则。优选的，探测模块中除了探测器，还包括其他对信号探测有益的器件，例如：当信号光传播到探测器感光面位置时的光斑过大以至于导致信号损失时，在探测器前方放置一聚焦透镜以缩小光斑；当信号光为荧光时，在探测器前方放置截止滤光片以避免荧光波段以外其他光的干扰；当空间中存在与该路信号光波长没有显著差异的其他非该路信号光干扰时，使用聚焦透镜以及小孔的组合，使该路信号光通过小孔后再进入探测器，以抑制干扰。
控制采集处理模块的功能包括：对所有的主动器件进行控制(主要是扫描模块中的扫描器，也包括其他模块中的主动器件)，使其在成像过程中工作；对探测器输出的信号进行采集，包括信号放大、滤波、模数转换等操作，使探测器输出的原始信号变为能够进行处理数据；对采集到的信号进行处理，包括拼接、灰度变换等操作，使采集到的数据以图像或其他数据形式展示或存储。
本发明实施例的并行多区域成像方法的成像过程为：照明光由光源模块1发出，经预处理模块2达到合适的光束直径、光功率或偏振态后，再进入多焦点生成模块3，多焦点生成模块3对照明光进行调制以产生两束或多束不同方向或聚焦程度的多路子光束，然后照明光继续传播至分光器4，经过分光器4后传输至扫描模块5，再经过中继光路6传播至物镜7，经物镜7在物方形成两个或多个焦点(或一维聚焦的照明线)；照明光照射到样品0上以后，在样品0上通过反射、散射或荧光等效应产生信号光；信号光被物镜7收集，经过中继光路6传播到扫描模块5上，扫描模块5在控制采集处理模块10的控制下进行扫描，信号光再经过扫描模块5传播至分光器4，分光器使信号光沿着与照明光不相同的空间路径继续传播，然后信号光进入空间解调模块8，空间解调模块8将各路照明子光束在样品上产生的各路信号光在空间上分离，探测模块9对分离后的各路信号光进行探测，控制采集处理模块10对探测模块9输出的电信号进行采集和处理并最终以图像或其他数据形式展示或存储。
上述结构和成像过程使用同一个物镜既进行照明又进行信号光的收集。如图2所示，本发明实施例还包括其他形式的结构和成像过程，例如使用两个物镜分别进行照明和信号光的收集。其结构的变动在于：扫描模块、中继光路和物镜的数目均为二，分光器置于第二扫描模块的后方。其成像过程为：照明光由光源模块1发出，经预处理模块2达到合适的光束直径、光功率或偏振态后，再进入多焦点生成模块3，多焦点生成模块3对照明光进行调制以产生两束或多束不同方向或聚焦程度的多路子光束，然后照明光继续传播至第一扫描模块5a，再经过第一中继光路6a传播至第一物镜7a，经第一物镜7a在物方形成两个或多个焦点(或一维聚焦的照明线)；照明光照射到样品0上以后，在样品0上通过反射、散射或荧光等效应产生信号光；信号光被第二物镜7b收集，经过第二中继光路6b传播到第二扫描模块5b上，第二扫描模块5b在控制采集处理模块10的控制下进行扫 描，照明光再经过第二扫描模块5b传播至分光器4，分光器4滤除掉和信号光混叠在一起的一部分照明光，然后信号光传播至空间解调模块8，空间解调模块8将各路照明子光束在样品上产生的各路信号光在空间上分离，探测模块9对分离后的各路信号光进行探测，控制采集处理模块10对探测模块9输出的电信号进行采集和处理并最终以图像或其他数据形式展示或存储。
对于单个物镜的结构，基于光路可逆原理，信号光经过扫描模块以后其空间位置不随扫描模块的扫描而变化，称为退扫描。对于两个物镜的结构，第二扫描模块5b与第一扫描模块5a的扫描过程要同步以实现推扫描的效果；分光器4的功能也有所变化，只需要将和信号光混叠在一起的照明光滤除即可。
本发明实施例的多焦点生成模块的具体实施方案1如图3所示。半波片31a置于偏振分光棱镜32a前方，偏振分光棱镜32a将输入光分为反射和透射两束，半波片31a用于调节偏振分光棱镜32a的反射光和透射光的能量比。半波片31b置于偏振分光棱镜32b前方，偏振分光棱镜32b将偏振分光棱镜32a的反射光分为两束，半波片31b用于调节偏振分光棱镜32b的反射光和透射光的能量比。利用偏振分光棱镜32a和32b的分光效果，最终得到了三路子光束。其中第一路子光束没有经过透镜的调制，第二路子光束经透镜34a和透镜34b的调制，第三路子光束经透镜34c和透镜34d的调制，调制后光束的方向和聚焦发散程度发生变化。第一路子光束经反射镜33a反射，第三路子光束经反射镜33b反射。第一路子光束经偏振分光棱镜32c透射，第二路子光束被偏振分光棱镜32c反射，第一路和第二路子光束合束，然后经半波片31c调制偏振态。半波片31c的作用是使第一路和第二路子光束合束后的光束取一个合适的偏振态，使其被偏振分光棱镜32d反射。第三路子光束经偏振分光棱镜32d透射。经过偏振分光棱镜32d后，三路子光束全部合束，此时虽然三路子光束的方向和发散聚焦程度不 尽相同，但是在空间中还是大量重叠。
本发明实施例的多焦点生成模块的具体实施方案2如图4所示。空间光调制器35在控制采集处理模块10的控制下产生特定调制图样。不同的调制图样可以生成不同的多焦点分布，具体来讲，随空间线性变化的相位调制分布用于改变出射光的方向，随空间以透镜调制函数形式分布的相位调制图样用于改变出射光的聚焦发散程度(透镜调制函数是圆对称的，其圆心位置也可以用来改变出射光的方向)。
本发明实施例的分光器的具体实施方案1如图5所示。二向色片41对不同波长的光具有不同的反射率和透过率，表现为长波长的光高透，对短波长的光高反，从而使得不同波长的光传播路径不同。
本发明实施例的分光器的具体实施方案2如图6所示。偏振分光棱镜42对不同偏振态的光具有不同的反射率和透过率，表现为对偏振态为p的光高透，对偏振态为s的光高反，从而使得不同偏振态的光传播路径不同。本发明实施例的分光器的具体实施方案3如图7所示。反射镜43的边缘置于两束光之间，使其对一路光反射，而另一路不受遮挡的继续传播，从而使得空间中传播路径不重叠的光进一步在空间中分开。
本发明实施例的空间解调模块的具体实施方案1如图8所示。带孔反射镜81的镜面中间有一个可透光的小孔。空间中有两路光，其中一路的焦点与带孔反射镜81的小孔重叠，从而这一路会穿过小孔继续传播，另一路的焦点与带孔反射镜81的小孔不重叠，从而这一路大部分能量会被反射镜反射。最终穿过小孔的光束80a与被反射的光束80b在空间中不再重叠。本发明实施例的空间解调模块的具体实施方案2如图9所示。数字微镜器件82在控制采集处理模块10的控制下产生特定调制图样。空间中有两路光，其中一路聚焦在数字微镜器件82的工作面上，数字微镜器件82上位于该路焦点位置的微镜偏转到一个状态，其他微镜偏转到另一个状态，于是聚焦到数字微镜器件82上的这一路光被数字微镜器件82反射到某个方 向，另一路光被反射到另一个方向，最终形成在空间中不重叠的光束80c和80d。
本发明实施例的空间解调模块的具体实施方案3如图10所示。空间光调制器83在控制采集处理模块10的控制下产生特定调制图样。空间中有三路光，它们的聚焦程度不同。空间光调制器83上的图样为变形光栅(离轴的菲涅尔波带片或离轴的全息透镜)，它与透镜831配合，将不同聚焦程度的三路光衍射到不同的衍射级次上，最终形成空间中不重叠的光束80e、80f和80g。
本发明实施例的整体结构的具体实施方案1如图11所示。飞秒激光器11发出的激光经透镜21a和透镜21b组成的预处理模块扩束后，射到空间光调制器35上，空间光调制器对波前进行调制生成两个焦点，多焦点光束经透镜351a和透镜351b中继后通过二向色镜41，再入射到由检流计镜51a和检流计镜51b组成的双轴扫描模块上，经扫描模块扫描后，再经中继透镜61a和中继透镜61b入射到物镜71上，经物镜71聚焦到样品0上。飞秒激光在样品内产生荧光，荧光信号经物镜71收集，沿路返回，最后射到二向色镜41上并被二向色镜41反射，反射后的光经透镜811聚焦后，一路光通过带孔反射镜81的小孔被光电倍增管91a所探测，另一路被带孔反射镜81反射并被光电倍增管91b探测。图中未画出控制采集处理模块，空间光调制器35、检流计镜51a和51b均受控制采集处理模块的控制，同时光电倍增管91a和91b的输出信号被控制采集处理模块采集。
本发明实施例的整体结构的具体实施方案2如图12所示。飞秒激光器11发出的激光经透镜21a和透镜21b组成的预处理模块扩束后，经半波片31a调整偏振态后入射到偏振分光棱镜321a上。偏振分光棱镜321a将光分成两路，一路经透镜34a和透镜34b调整会聚发散程度并入射到偏振分光棱镜321b上，另一路经反射镜331a和331b反射后也入射到偏振分光棱镜321b上，偏振分光棱镜321b将两路光合束。合束的光经反射镜331c反射 后，通过透镜351a和透镜351b的中继，经二向色镜41后入射到由检流计镜51a和检流计镜51b组成的双轴扫描模块上，经扫描模块扫描后，再经中继透镜61a和中继透镜61b入射到物镜71上，经物镜71聚焦到样品0上。飞秒激光在样品内产生荧光，荧光信号经物镜71收集，沿路返回，最后射到二向色镜41上并被二向色镜41反射，反射后的光经透镜811聚焦后，一路光通过带孔反射镜81的小孔被光电倍增管91a所探测，另一路被带孔反射镜81反射并被光电倍增管91b探测。图中未画出控制采集处理模块，检流计镜51a和51b均受控制采集处理模块的控制，同时光电倍增管91a和91b的输出信号被控制采集处理模块采集。
本发明实施例的整体结构的具体实施方案3如图13所示。飞秒激光器11发出的激光经透镜21a和透镜21b组成的预处理模块扩束后，入射到反射光栅361上，反射光栅361用于对数字微镜器件36的空间色散进行补偿，反射光栅361的出射光经二向色镜41后入射到数字微镜器件36上，数字微镜器件36同时实现多焦点生成和扫描的功能。光通过数字微镜器件36后，经中继透镜61a和61b入射到物镜71上，物镜71将光聚焦到焦点上。飞秒激光在样品内产生荧光，荧光信号经物镜71收集，沿路返回，最后射到二向色镜41上并被二向色镜41反射，反射后的光经色散补偿模块49补偿，再经透镜811聚焦后，一路光通过带孔反射镜81的小孔被光电倍增管91a所探测，另一路被带孔反射镜81反射并被光电倍增管91b探测。图中未画出控制采集处理模块，数字微镜器件36受控制采集处理模块的控制，同时光电倍增管91a和91b的输出信号被控制采集处理模块采集。
本发明实施例的整体结构的具体实施方案4如图14所示。飞秒激光器11发出的激光经透镜21a和透镜21b组成的预处理模块扩束后，入射到透射光栅352上，透射光栅352用于对空间光调制器35的空间色散进行补偿，透射光栅352的出射光射到空间光调制器35上，空间光调制器对波前进行调制生成两个焦点，多焦点光束经透镜351a和透镜351b中继后通过二向 色镜41，再入射到由声光偏转器52a和声光偏转器52b组成的双轴扫描模块上，经扫描模块扫描后，再经中继透镜61a和中继透镜61b入射到物镜71上，经物镜71聚焦到样品0上。飞秒激光在样品内产生荧光，荧光信号经物镜71收集，沿路返回，最后射到二向色镜41上并被二向色镜41反射，反射后的光经色散补偿模块49补偿，再经透镜811聚焦后，一路光通过带孔反射镜81的小孔被光电倍增管91a所探测，另一路被带孔反射镜81反射并被光电倍增管91b探测。图中未画出控制采集处理模块，空间光调制器35、声光偏转器52a和52b均受控制采集处理模块的控制，同时光电倍增管91a和91b的输出信号被控制采集处理模块采集。
本发明实施例的整体结构的具体实施方案5如图15所示。飞秒激光器11发出的激光经透镜21a和透镜21b组成的预处理模块扩束后，入射到透射光栅352上，透射光栅352用于对空间光调制器35的空间色散进行补偿，透射光栅352的出射光射到空间光调制器35上，空间光调制器对波前进行调制生成两个焦点，多焦点光束经透镜351a和透镜351b中继后通过二向色镜41，再入射到由检流计镜51a和检流计镜51b组成的双轴扫描模块上，经扫描模块扫描后，再经中继透镜61a和中继透镜61b入射到物镜71上，经物镜71聚焦到样品0上。飞秒激光在样品内产生荧光，荧光信号经物镜71收集，沿路返回，最后射到二向色镜41上并被二向色镜41反射，反射后的光入射到数字微镜器件83上，数字微镜器件83上产生变形光栅(离轴的菲涅尔波带片)，使一路光聚焦到小孔381a上然后被光电倍增管91a所探测，另一路聚焦到小孔831b上然后被光电倍增管91b探测。图中未画出控制采集处理模块，空间光调制器35、检流计镜51a和51b、数字微镜器件83均受控制采集处理模块的控制，同时光电倍增管91a和91b的输出信号被控制采集处理模块采集。
本发明实施例的整体结构的具体实施方案6如图16所示。飞秒激光器11发出的激光经透镜21a和透镜21b组成的预处理模块扩束后，入射到透 射光栅352上，透射光栅352用于对空间光调制器35的空间色散进行补偿，透射光栅352的出射光射到空间光调制器35上，空间光调制器对波前进行调制生成多个焦点，多焦点光束经透镜351a和透镜351b中继后通过二向色镜41，再入射到由检流计镜51a和检流计镜51b组成的双轴扫描模块上，经扫描模块扫描后，再经中继透镜61a和中继透镜61b入射到物镜71上，经物镜71聚焦到样品0上。飞秒激光在样品内产生荧光，荧光信号经物镜71收集，沿路返回，最后射到二向色镜41上并被二向色镜41反射，反射后的光经透镜841a和透镜841b中继后入射到调制器84上。调制器84的调制函数为空间坐标的n次函数(n≥2)，可表示为
Figure PCTCN2018106072-appb-000001
其中A为强度调制函数且默认为实常数，a和b为实常数，当n＝2时，调制器表现为透镜，当n＝3时，调制器84的出射光表现为艾利光束(Airy beam)。调制器84为被动型调制器件，由透明材料加工而成。调制器84也可以为空间光调制器或数字微镜器件。当调制器84为数字微镜器件时，其图样为所述调制函数的二值化结果，例如对相位取余弦，然后按照阈值进行二值化。光经过调制器84后，表现出聚焦的特性。若空间光调制器35产生的多个焦点的轴向位置相同，那么n＝2时，即调制器84为透镜时，即可使不同焦点聚焦在同一平面上且横向位置不同。若空间光调制器35产生的多个焦点存在轴向位置不同的情况，直接使用透镜进行聚焦，离焦光斑和聚焦光斑会存在重叠，无法进行空间解调，因此取n>2，此时调制器84后方聚焦光斑的焦深大于同等数值孔径下透镜焦点的焦深，消除了离焦光斑和聚焦光斑重叠的现象，且不同焦点聚焦后的横向位置不同。不同焦点均聚焦在同一平面且横向位置不同，即实现了空间解调。探测器92位于调制器84后方的聚焦区域内，此时信号光在探测器92上表现为横向位置不同的聚焦光斑。探测器92为阵列探测器，例如多阳极光电倍增管、阵列雪崩二极管、面阵CMOS传感器、面阵CCD传感器等，从而可采集到不同聚焦光斑的信 号。图中未画出控制采集处理模块，空间光调制器35、检流计镜51a和51b以及其他主动型器件均受控制采集处理模块的控制，同时探测器92的输出信号被控制采集处理模块采集。
本领域的技术人员容易理解，以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。

Claims (6)

  1. 一种并行多区域成像装置，其特征在于，包括：多焦点生成模块、空间解调模块及探测模块；
    所述多焦点生成模块，用于对照明光进行调制，以在物方产生多个焦点，形成多个不同的照明区域，进而通过待成像样品产生多路信号光；
    所述空间解调模块，用于使各路信号光在空间上的能量分布不再重叠或重叠区域小于目标要求；
    所述探测模块，用于对经所述空间解调模块后的各路信号光进行独立探测，以实现并行多区域成像。
  2. 根据权利要求1所述的装置，其特征在于，在所述多焦点生成模块和所述空间解调模块之间，所述装置还包括分光器；
    所述分光器，用于将照明光和由待成像样品返回的信号光在空间上分离，以使所述多焦点生成模块输出的照明光传播至所述待成像样品，同时使自所述待成像样品返回的信号光传播至所述空间解调模块。
  3. 根据权利要求2所述的装置，其特征在于，所述分光器的分光方法包括按照波长分离、按照偏振态分离、按照空间位置分离及衰减式分离。
  4. 根据权利要求3所述的装置，其特征在于，在所述分光器之后，所述装置还包括扫描模块；
    所述扫描模块，用于改变光束的传播方向、发散程度或汇聚程度，使得物镜物方的焦点或一维聚焦的线在空间中改变位置，从而进行扫描。
  5. 根据权利要求4所述的装置，其特征在于，所述扫描模块中的扫描器件包括反射式、透射式或衍射式。
  6. 根据权利要求1至5任意一项所述的装置，其特征在于，在所述扫描模块之后，所述装置还包括：中继光路；
    所述中继光路，用于实现所述扫描模块与物镜的光瞳匹配，以在所述扫描模块的目标扫描范围内使得照明光均能进入到所述物镜中从而照明到所述待成像样品上。
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