CN114986911A - Biological micro-scaffold hardness visualization method and system, electronic equipment and storage medium - Google Patents
Biological micro-scaffold hardness visualization method and system, electronic equipment and storage medium Download PDFInfo
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Abstract
The embodiment of the invention discloses a method, a system, electronic equipment and a storage medium for visualizing the hardness of a biological micro-scaffold, and relates to the technical field of biological tissue construction, wherein the method comprises the following steps: digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram; transforming and filtering the digital hologram to obtain a spectrogram; calculating and obtaining a wrapping phase according to the obtained spectrogram; unwrapping the wrapped phase to obtain the refractive index of the target object; the method provided by the invention establishes the mapping relation between the local rigidity and the refractive index according to the relation between the local refractive index of the target and the cross-linked network density of the target. Indirect visualization and sampling of real-time rigidity are realized by observing the change of the refractive index of the structure, so that the real-time observation of the rigidity of the cell micro-scaffold is realized.
Description
Technical Field
The invention relates to the technical field of biological tissue construction, in particular to a method and a system for visualizing the hardness of a biological micro-scaffold, electronic equipment and a storage medium.
Background
Extracellular matrix (ECM) is an important component of biological tissue, providing an important mechanical support environment for the life of cell populations. The range of mechanical stiffness for different human tissues usually spans a large extent. According to measurements, the Young's modulus of prostate tissue is about 90kPa, whereas that of breast tissue is only 8.1 kPa. Different mechanical stiffness of different tissues can induce different physiological expressions (e.g., secretion and interaction) of cell populations, thereby exhibiting different biological functions on a macroscopic scale. In addition, subtle stiffness changes between local micro-scale regions within a single tissue can also lead to differences in behavior at the cellular level. For example, mouse fibroblasts (3T3 cells) can migrate from a soft region to a rigid region, with neurons synapse more branches on softer substrates. Therefore, in designing an ECM, it is necessary not only to be able to adjust the mechanical stiffness over a large span, but also to precisely control the gradient change in stiffness.
Biocompatible polymers (e.g., uv light curable hydrogels) are often used to design biomimetic extracellular matrices in vitro. Therefore, 3D bio-printing technologies such as 3D laser lithography, photo-patterning, and Digital Light Processing (DLP) have emerged. These bioprinting techniques have focused primarily on optimization of the printed shape, and have now made breakthrough advances in the accurate reproduction of the 3D morphology of natural tissue topographically.
However, dynamically modulating the mechanical properties of biomimetic ECM at the microscopic scale remains a significant challenge due to the lack of ability to effectively detect mechanical stiffness in real time. An Atomic Force Microscope (AFM) is used as a mechanical measuring tool, the mechanical properties of cells and other biological materials can be flexibly determined, and the measuring precision of the AFM depends on the size of nano-indentation. However, AFM, as a complex device, is difficult to combine with existing 3D bioprinting systems in order to obtain data of young's modulus as feedback in real time during ECM manufacturing to adjust mechanical properties at its microscopic scale.
Disclosure of Invention
The embodiment of the invention aims to provide a method and a system for visualizing the hardness of a biological micro-scaffold, electronic equipment and a storage medium, which are used for solving the problem that the hardness condition of biological tissues cannot be observed in real time in the life printing process so as to be properly adjusted in the prior art.
To achieve the above object, the contents of the embodiments of the present invention are further illustrated by five aspects:
in a first aspect, a method for visualizing the hardness of a microbial scaffold is provided, the method comprising the following steps:
digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
transforming and filtering the digital hologram to obtain a spectrogram;
calculating and obtaining a wrapping phase according to the obtained spectrogram;
unwrapping the wrapped phase to obtain the refractive index of the target object;
and obtaining the hardness of the target object according to the mapping relation between the refractive index and the Young modulus of the target object.
With reference to the first aspect, a method for obtaining a digital hologram by digitally sampling a hologram formed by superimposing acquired reference light and object light transmitted through a target object includes the steps of;
respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
and digitally sampling the hologram through a photoelectric converter to obtain a digital hologram.
With reference to the first aspect, the method for transforming and filtering the digital hologram to obtain a spectrogram comprises the following steps:
carrying out Fourier transform on the digital hologram to obtain a spectrogram;
filtering out parts except the +1 level image in the spectrogram by a high-pass filter;
and obtaining a filtered spectrogram by performing inverse Fourier transform on the part reserved after the filtering.
With reference to the first aspect, the method for calculating the wrapped phase from the obtained spectrogram includes the following steps:
respectively calculating values of a real number part and an imaginary number part of the spectrogram;
and comparing the value of the real number part obtained by calculation with the value of the imaginary number part, and then taking the value as the input of an arc tangent function to obtain the wrapping phase of the spectrogram by calculation.
With reference to the first aspect, the method for unwrapping the wrapped phase to obtain the refractive index of the object includes the following steps:
reducing and denoising the wrapped phase by adopting a least square method so as to obtain a real phase;
and calculating according to the real phase and the thickness of the target object to obtain the refractive index of the target object.
In a second aspect, there is provided a biological micro-stent stiffness visualization system, the system comprising:
a sampling module: digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
a data processing module: transforming and filtering the digital hologram to obtain a spectrogram;
a calculation module: calculating and obtaining a wrapping phase according to the obtained spectrogram;
an unwrapping module: unwrapping the wrapped phase to obtain the refractive index of the target object;
an operation module: and obtaining the hardness of the target object according to the mapping relation between the refractive index and the Young modulus of the target object.
With reference to the second aspect, the specific operation process of the sampling module includes: respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
and digitally sampling the hologram through a photoelectric converter to obtain a digital hologram.
In a third aspect, there is provided a biological micro-stent printing system comprising a biological micro-stent stiffness visualization system, the apparatus comprising:
a data acquisition module: the shape of the microbial scaffold and the hardness data of each part are introduced;
an initialization module: the method is used for initializing parameter data, defining a hardness threshold value and preparing a material required by hardening;
a curing module: the device is used for emitting curing light to the curing material according to the shape of the biological micro-scaffold and curing the curing material according to the hardness data;
biological little support hardness visual system: the system is used for emitting object light and reference light and obtaining a curing value of a curing material according to interference calculation formed by the object light and the reference light;
a judging module: and the device is used for comparing the curing value with a hardness threshold value, judging the hardening degree and adjusting the intensity of curing light according to the hardening degree.
In a fourth aspect, there is provided an electronic device comprising a memory, a processor and a computer program stored in the memory and executable on the processor, wherein the processor executes the computer program to perform the steps of the method for visualizing stiffness of a biological micro-stent according to the first aspect.
In a fifth aspect, there is provided a non-transitory computer readable storage medium having stored thereon a computer program which, when executed by a processor, performs the steps of the method for visualizing stiffness of a biological micro-stent according to the first aspect.
The embodiment of the invention has the following advantages: and establishing a mapping relation between the local rigidity and the refractive index according to the relation between the local refractive index of the target object and the cross-linked network density of the target object. Indirect visualization and sampling of real-time rigidity are realized by observing the change of the refractive index of the structure, so that the real-time observation of the rigidity of the cell micro-scaffold is realized. Meanwhile, the hardness control in the process of printing the cell micro-scaffold is realized according to the hardness condition.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
The structures, ratios, sizes, and the like shown in the present specification are only used for matching with the contents disclosed in the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions that the present invention can be implemented, so that the present invention has no technical significance, and any structural modifications, changes in the ratio relationship, or adjustments of the sizes, without affecting the effects and the achievable by the present invention, should still fall within the range that the technical contents disclosed in the present invention can cover.
Fig. 1 is a schematic diagram of a method provided in an embodiment of the present invention.
Fig. 2 is a schematic flow chart of a method according to an embodiment of the present invention.
Fig. 3 is a schematic operation diagram of a method according to an embodiment of the present invention.
Fig. 4 is a schematic flow chart of a printing system according to an embodiment of the present invention.
Fig. 5 is a hologram photographed by the CCD camera according to the embodiment of the present invention.
Fig. 6 is a schematic diagram of a hologram in the wrapped phase obtained after the processing in fig. 5 according to an embodiment of the present invention.
FIG. 7 is a schematic diagram of the unwrapped hologram of FIG. 6 according to an embodiment of the present invention.
FIG. 8 is a schematic representation of FIG. 7 after reduction in an embodiment of the present disclosure.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention. Furthermore, the terms "first", "second", etc. are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first," "second," etc. may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood by those of ordinary skill in the art through specific situations.
The invention takes hydrogel as a printing material, controls the hardness of the hydrogel by adjusting the emergent intensity of ultraviolet light, takes the ultraviolet light curing hydrogel as a typical polymer, and the structure density of the ultraviolet light curing hydrogel can be characterized by cross-linked network density which is determined by the degree of photocuring. The local refractive index of the hydrogel may reflect its own crosslink network density, thus allowing a mapping between local stiffness and refractive index. Therefore, by observing the change of the refractive index of the structure, indirect visualization and sampling of the real-time rigidity can be realized. In order to measure the refractive index of a structure, techniques such as a Precision Goniometer (PG), an Abbe Refractometer (AR), and a Digital Holographic Microscope (DHM) are generally used. Among these techniques, Digital Holographic Microscopy (DHM) offers the best solution for measuring refractive index, with the advantages of real-time, in-situ and flexible detection. DHM is expected to monitor, sample, and feed back during 3D printing to adjust the local mechanical properties of the ECM.
The following further illustrates the practice of the invention in five respects:
a method for visualizing the stiffness of a microbial scaffold, the method comprising the steps of:
s1: digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
the concrete implementation steps comprise:
respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
carrying out digital sampling on the hologram through a photoelectric converter to obtain a digital hologram;
in holography, the recording intensity G (x, y) of a hologram is a square area where the amplitudes of the object and reference waves are superimposed, as follows:
G(x，y)＝|O(x，y)| 2 +|R(x，y)| 2 +O(x，y)R*(x，y)+O*(x，y)R(x，y) (1)
wherein, O (x, y) and R (x, y) are the light field intensity of the object light and the reference light respectively, "+" represents conjugate complex number, the first two terms are zero order terms which represent the intensity of the reference light and the object light, therefore, the first two terms do not provide the space information of the optical field of the object, the third term and the fourth term provide the space frequency of the hologram recorded by the CCD camera and correspond to the virtual image and the real image respectively, and the light intensity is photoelectrically converted by the CCD to obtain the digital hologram;
the following formula:
G H (x，y)＝w(x，y)G(x，y) (2)
where w (x, y) is a window function of the photoelectric conversion device, the continuous function G (x, y) is converted into a theoretical value G by equation (2) H (x，y)；
As shown in fig. 3, since the detection plane of the CCD camera is composed of discrete pixel points, the intensity G (x, y) continuously distributed in the hologram is digitally sampled by the CCD camera to form discrete grid regions; thus, the digital hologram may be implemented using an M N matrix G M，N A representation in which each element is defined as:
G M，N [m，n]＝G H (mΔx，nΔy) (3)
wherein M is 1, 2, …, M, N is 1, 2, …, N; Δ x and Δ y are the individual pixel sizes of the CCD camera;
s2: transforming and filtering the digital hologram to obtain a spectrogram;
the concrete implementation steps comprise:
carrying out Fourier transform on the digital hologram to obtain a spectrogram;
filtering out parts except the +1 level image in the spectrogram by a high-pass filter;
obtaining a filtered spectrogram by performing inverse Fourier transform on the part reserved after filtering;
as shown in the spectrogram of fig. 3, when the spectrogram is obtained by fourier transform of a digital hologram, images of level +1, level 0 and level-1 are obtained, so that the image of level +1 is retained and the image of level 0 and level-1 is filtered out by a high-pass filter;
based on frequency domain filtering, the digital hologram is fourier-transformed, and the fourier transform is obtained by the following equations (1) and (2):
F{G H (x，y)}＝F{w(x，y)|O| 2 }+F{w(x，y)|R| 2 }+F{w(x，y)R*O} (4)+F{w(x，y)RO*}
≈F{|O| 2 }+F{|R| 2 }+F{R*O}+F{RO*}
wherein F { | O- 2 Y and F { | R- 2 The components F { R x O } + F { RO } are clearly separated from the high frequency components F { R x O } + F { RO } in the spectrogram, so that a suitable high pass filter can be selected
Making F { | O- 2 Y and F { | R- 2 And f, filtering:
H(ζ，η)[F{|O| 2 }+F{|R| 2 }+F{R*O}+F{RO*}]≈F{R*O}+F{RO*}(5)
the spectrum a at the plane Z ═ 0 available from (4) and (5) M，N (ζ, η; O) is:
A M，N (ζ，η；0)＝H[F{G M，N }] (6)
f { } represents Fourier transform, and transforms the obtained time domain hologram into a frequency domain space and filters out clutter so as to obtain a clear restored hologram; ξ and η are the spatial frequencies in the M and N directions, respectively;
s3: calculating and obtaining a wrapping phase according to the obtained spectrogram;
the concrete implementation steps comprise:
respectively calculating values of a real number part and an imaginary number part of the spectrogram;
comparing the value of the real number part obtained by calculation with the value of the imaginary number part, and then taking the value of the real number part and the value of the imaginary number part as the input of an arc tangent function, and obtaining the wrapping phase of the spectrogram by calculation;
the specific operation process is as follows:
according to the Fresnel diffraction integral expression:
the rewritables are:
wherein FFT represents fast fourier transform; i.e. the frequency spectrum a at the plane Z ═ d M，N (ζ, η; d) can be calculated by the formula (8):
wherein λ is the wavelength; d is the propagation distance of the object wave.
Then, by mixing A M，N (ζ, η; d) inverse Fourier transform to obtain a hologram reproduced after frequency domain filtering:
U d M,N ＝F -1 {A M,N (ζ,η；d)} (10)
wherein F -1 { } denotes an inverse fourier transform.
Phase distribution of sample
The parameters of the reconstructed complex amplitude wavefront can be obtained from a single digital hologram by calculating:
where Im () and Re () represent coefficients of real and imaginary parts.
S4: unwrapping the wrapped phase to obtain the refractive index of the target object;
the concrete implementation steps comprise:
reducing and denoising the wrapped phase by adopting a least square method so as to obtain a real phase;
calculating according to the real phase and the thickness of the target object to obtain the refractive index of the target object;
as the arctangent function is used in the formula (11), the main value domain is (-pi, pi), so that the bit image is limited within (-pi, pi), and the directly calculated phase can be 'truncated', so that the unwrapping program needs to be written to restore the real morphology of the phase;
reducing and denoising the wrapped phase by a least square method to obtain a real phase phi M，N (ii) a Thus, the refractive index of the sample is:
wherein H M，N Is the thickness of the sample.
S5: obtaining the hardness of the target object according to the mapping relation between the refractive index and the Young modulus of the target object;
according to the pre-completed parameter calibration experiment, the hardness of the printed micro-support can be visualized in real time according to a mapping relation formula between the Young modulus and the refractive index:
E M，N ＝aL M，N +b (13)
wherein E M，N Is the Young's modulus of the sample. a and b are coefficients obtained after curve fitting;
the method realizes the hardness control of the cell micro-scaffold in the printing process, the control range of the hardness (Young modulus) of the GelMA material used for the experiment is 10kPa to 50kPa, and the regulation and control precision is 2.9 kPa.
According to a method of an embodiment of the present invention, there is provided a biological micro-stent stiffness visualization system, the system comprising:
a sampling module: digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
the specific operation process of the sampling module comprises the following steps: respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
carrying out digital sampling on the hologram through a photoelectric converter to obtain a digital hologram;
a data processing module: transforming and filtering the digital hologram to obtain a spectrogram;
the data processing module comprises the following concrete implementation steps: carrying out Fourier transform on the digital hologram to obtain a spectrogram;
filtering out parts except the +1 level image in the spectrogram by a high-pass filter;
obtaining a filtered spectrogram by performing inverse Fourier transform on the part reserved after filtering;
a calculation module: calculating and obtaining a wrapping phase according to the obtained spectrogram;
the specific implementation steps of the computing module are as follows: respectively calculating values of a real number part and an imaginary number part of the spectrogram;
comparing the value of the real number part obtained by calculation with the value of the imaginary number part, and then taking the value of the real number part and the value of the imaginary number part as the input of an arc tangent function, and obtaining the wrapping phase of the spectrogram by calculation;
an unwrapping module: unwrapping the wrapped phase to obtain the refractive index of the target object;
the specific implementation steps of the unpacking module are as follows: reducing and denoising the wrapped phase by adopting a least square method so as to obtain a real phase;
calculating according to the real phase and the thickness of the target object to obtain the refractive index of the target object;
an operation module: obtaining the hardness of the target object according to the mapping relation between the refractive index and the Young modulus of the target object;
according to a pre-completed parameter calibration experiment, the hardness of the printed micro-stent can be visualized in real time according to a mapping relation formula between the Young modulus and the refractive index:
E M，N ＝aL M，N +b
wherein E M，N Is the Young's modulus of the sample. and a and b are coefficients obtained after curve fitting.
According to the method of the embodiment of the invention, a biological micro-stent printing system is provided, which comprises a biological micro-stent hardness visualization system, and the device comprises:
a data acquisition module: the shape and the hardness data of each part for leading in the microbial scaffold;
inputting the shape of the target object to be printed and the hardness data of each part into a printing system, wherein the system can be connected with an upper computer to store and analyze the data;
an initialization module: the method is used for initializing parameter data, defining a hardness threshold value and preparing a material required by hardening;
planning a printing step by analyzing the shape and the hardness of a target object, defining a hardness threshold according to hardness data, and filling hydrogel required for hardening into a micro flow channel;
a curing module: the device is used for emitting curing light to the curing material according to the shape of the biological micro-scaffold and curing the curing material according to the hardness data;
the curing material is a hydrogel material, and the curing light is ultraviolet light or visible light; the hydrogel material has excellent biocompatibility, and forms a three-dimensional structure which is suitable for cell growth and differentiation and has certain strength after ultraviolet light or visible light excitation curing reaction; after preparing the hydrogel and the curing light, adjusting the emergent shape and the intensity of the ultraviolet light according to the shape and the hardness data of the target object to realize the curing of the hydrogel material;
biological little support hardness visual system: the system is used for emitting object light and reference light and obtaining a curing value of a curing material according to interference calculation formed by the object light and the reference light;
the system comprises:
a sampling module: digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
the specific operation process of the sampling module comprises the following steps: respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
carrying out digital sampling on the hologram through a photoelectric converter to obtain a digital hologram;
a data processing module: transforming and filtering the digital hologram to obtain a spectrogram;
the data processing module comprises the following concrete implementation steps: carrying out Fourier transform on the digital hologram to obtain a spectrogram;
filtering out parts except the +1 level image in the spectrogram by a high-pass filter;
obtaining a filtered spectrogram by performing inverse Fourier transform on the part reserved after filtering;
a calculation module: calculating and obtaining a wrapping phase according to the obtained spectrogram;
the specific implementation steps of the computing module are as follows: respectively calculating values of a real number part and an imaginary number part of the spectrogram;
comparing the value of the real number part obtained by calculation with the value of the imaginary number part, and then taking the value of the real number part and the value of the imaginary number part as the input of an arc tangent function, and obtaining the wrapping phase of the spectrogram by calculation;
an unwrapping module: unwrapping the wrapped phase to obtain the refractive index of the target object;
the specific implementation steps of the unpacking module are as follows: reducing and denoising the wrapped phase by adopting a least square method so as to obtain a real phase;
calculating according to the real phase and the thickness of the target object to obtain the refractive index of the target object;
an operation module: obtaining the hardness of the target object according to the mapping relation between the refractive index and the Young modulus of the target object;
according to the pre-completed parameter calibration experiment, the hardness of the printed micro-support can be visualized in real time according to a mapping relation formula between the Young modulus and the refractive index:
E M，N ＝aL M，N +b
wherein E M，N Is the Young's modulus of the sample. a and b are coefficients obtained after curve fitting;
a judging module: the device is used for comparing the curing value with a hardness threshold value, judging the hardening degree and adjusting the intensity of curing light according to the hardening degree;
comparing the obtained hardness value with a threshold value, judging whether the hardness reaches the threshold value, and weakening the irradiation intensity of the ultraviolet light source to irradiate the next layer when the curing intensity reaches the threshold value;
when the curing intensity is uneven, adjusting the ultraviolet shape of the part with the curing intensity reaching the threshold value, and continuing to irradiate the part which does not reach the threshold value until the part reaches the threshold value;
when the curing intensity reaches a certain threshold value, the ultraviolet light source can be controlled to weaken the emitted ultraviolet light.
According to the method of the embodiment of the present invention, there is provided an electronic device comprising a memory, a processor and a computer program stored in the memory and executable on the processor, wherein the processor executes the computer program to realize the steps of the method for visualizing the hardness of a biological micro-scaffold described in S1 to S5.
According to the method of an embodiment of the present invention, there is provided a non-transitory computer readable storage medium having stored thereon a computer program, wherein the computer program when executed by a processor implements the steps of the method for visualizing stiffness of a microbial micro-stent described in S1 to S5.
Although the invention has been described in detail with respect to the general description and the specific embodiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A method for visualizing the hardness of a microbial scaffold, which is characterized by comprising the following steps:
digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
transforming and filtering the digital hologram to obtain a spectrogram;
calculating and obtaining a wrapping phase according to the obtained spectrogram;
unwrapping the wrapped phase to obtain the refractive index of the target object;
and obtaining the hardness of the target object according to the mapping relation between the refractive index and the Young modulus of the target object.
2. The method for visualizing the hardness of a microbial scaffold according to claim 1, wherein: the method for obtaining the digital hologram by digitally sampling the hologram formed by overlapping the acquired reference light and the object light transmitted through the target object comprises the following steps;
respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
and digitally sampling the hologram through a photoelectric converter to obtain a digital hologram.
3. The method for visualizing the hardness of a microbial scaffold according to claim 2, wherein: the method for transforming and filtering the digital hologram to obtain the spectrogram comprises the following steps:
carrying out Fourier transform on the digital hologram to obtain a frequency spectrum;
filtering out parts except the +1 level image in the spectrogram by a high-pass filter;
and obtaining a filtered spectrogram by performing inverse Fourier transform on the part reserved after the filtering.
4. The method for visualizing the hardness of a microbial scaffold according to claim 3, wherein: the method for calculating the wrapping phase according to the acquired spectrogram comprises the following steps:
respectively calculating values of a real number part and an imaginary number part of the spectrogram;
and comparing the value of the real number part obtained by calculation with the value of the imaginary number part, and then taking the value as the input of an arc tangent function to obtain the wrapping phase of the spectrogram by calculation.
5. The method for visualizing the hardness of a biological micro-stent as claimed in claim 4, wherein: the method for unwrapping the wrapped phase to obtain the refractive index of the target object comprises the following steps:
reducing and denoising the wrapped phase by adopting a least square method so as to obtain a real phase;
and calculating according to the real phase and the thickness of the target object to obtain the refractive index of the target object.
6. A biological micro-scaffold hardness visualization system is characterized in that: the system comprises:
a sampling module: digitally sampling a hologram formed by overlapping the acquired reference light and object light penetrating through a target object to obtain a digital hologram;
a data processing module: transforming and filtering the digital hologram to obtain a spectrogram;
a calculation module: calculating and obtaining a wrapping phase according to the obtained spectrogram;
an unwrapping module: unwrapping the wrapped phase to obtain the refractive index of the target object;
an operation module: and obtaining the hardness of the target according to the mapping relation between the refractive index and the Young modulus of the target.
7. The system for visualizing the hardness of a biological micro-stent according to claim 6, wherein: the specific operation process of the sampling module comprises the following steps: respectively acquiring reference light and object light, wherein the object light and the reference light are polarized light with the same direction, and the object light and the reference light are superposed to generate light interference;
superposing the reference light and the object light to obtain a hologram;
and digitally sampling the hologram through a photoelectric converter to obtain a digital hologram.
8. The utility model provides a little support printing system of living beings, includes little support hardness visual system of living beings, its characterized in that: the apparatus comprises:
a data acquisition module: the shape of the microbial scaffold and the hardness data of each part are introduced;
an initialization module: the method is used for initializing parameter data, defining a hardness threshold value and preparing a material required by hardening;
a curing module: the device is used for emitting curing light to the curing material according to the shape of the biological micro-scaffold and curing the curing material according to the hardness data;
biological little support hardness visual system: the system is used for emitting object light and reference light and obtaining a curing value of a curing material according to interference calculation formed by the object light and the reference light;
a judging module: and the device is used for comparing the curing value with a hardness threshold value, judging the hardening degree and adjusting the intensity of curing light according to the hardening degree.
9. An electronic device comprising a memory, a processor and a computer program stored on the memory and executable on the processor, wherein the processor when executing the computer program implements the steps of the method for visualizing stiffness of a biological micro-scaffold according to any of claims 1 to 5.
10. A non-transitory computer readable storage medium having stored thereon a computer program, wherein the computer program, when executed by a processor, implements the steps of the method for visualizing stiffness of a biological micro-stent as in any one of claims 1 to 5.
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