CN114324244A - Optical detection method and system for orientation of biological membrane collagen beam based on weak coherent interference - Google Patents
Optical detection method and system for orientation of biological membrane collagen beam based on weak coherent interference Download PDFInfo
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Abstract
The invention discloses a method and a system for detecting the orientation of a biological film collagen beam based on weak coherent interference. The method comprises the steps of obtaining a biological film interference spectrum by using an optical coherent imaging system, carrying out Fourier transform on the interference spectrum to obtain a depth domain intensity signal, calculating a biological film surface coordinate based on the depth domain intensity signal, then fitting a biological film surface curve by using a polynomial, carrying out coordinate transform on the biological film according to the fitting curve to enable the surface of the biological film to tend to be horizontal, carrying out intensity signal projection of different depths on the flattened biological film, and displaying the orientation characteristic of a biological film collagen bundle by using an intensity signal projection diagram. The invention can realize one-time imaging to obtain the biomembrane collagen bundle orientation with depth resolution (micrometer precision), and can realize rapid evaluation and cutting of the biomembrane edge to obtain high-quality artificial heart biological valve leaflets.
Description
Technical Field
The invention relates to the orientation detection of collagen bundles of bioprosthetic tissue for implants, and more particularly to an optical nondestructive detection method of collagen bundle orientation of a biological membrane for fabrication of a bioprosthetic valve leaflet of a prosthetic heart.
Background
The normal native heart includes the aortic, mitral, tricuspid, and pulmonary valves, each of which has unidirectional leaflets to control the directional flow of blood through the heart. When the heart valve is diseased or damaged, in order to work normally, the heart valve can be replaced by a heart valve prosthesis.
The artificial heart biological valve is made of biological tissue materials, such as bovine pericardium tissue and porcine pericardium tissue which are used for manufacturing single leaflets of the artificial heart biological valve. A typical procedure commonly used to prepare bioprosthetic heart valve leaflets is to take a fresh animal pericardial sac, cut the pericardial sac flat and remove excess fat and impurities. The tissue is cross-linked with the drug after trimming the apparently unusable area, followed by removal of the rough edges of the tissue. The treated biofilm contains primarily collagen fibers, appearing as the outer surface of the fibers and the smooth inner surface. The biofilm is used to make leaflets of a bioprosthetic heart valve. The strength of leaflets made from a cut of biofilm depends on the average orientation of the collagen bundles, and optimizing the leaflets to be thinner based on collagen bundle orientation is also advantageous for interventional delivery of conventional heart valves.
At present, the optical detection method for the orientation of a collagen bundle of a biological film is to utilize polarized light to pass through the biological film, emergent light passes through a polarizer, and finally a stripe pattern with a direction is obtained on a detector plate, namely the average orientation of the collagen bundle. Although this method is also a non-destructive test, it requires continuous rotation of the polarizer direction to obtain the average orientation of the collagen bundles, which takes a long time, and it is impossible to obtain the orientation distribution of the collagen bundles at different depths in the biological membrane.
There is an urgent need for accurate and efficient detection and evaluation of the orientation of collagen bundles in biological membranes to obtain better quality leaflets of artificial heart valves.
Disclosure of Invention
The invention aims to provide a method and a system for detecting the orientation of a biological membrane collagen beam based on weak coherent interference aiming at the defects of the prior art. The method comprises the steps of acquiring a biological membrane interference spectrum by using an optical coherence imaging system (OCT), and carrying out Fourier transform on the interference spectrum to obtain a depth domain intensity signal. And calculating the surface coordinate of the biological membrane based on the depth domain intensity signal, fitting a surface curve of the biological membrane by using a polynomial, and performing coordinate transformation on the biological membrane according to the fitting curve to enable the surface of the biological membrane to tend to be horizontal. The flattened biological membrane can be subjected to intensity signal projection at different depths, and an intensity signal projection diagram can show the orientation characteristics of the collagen bundles of the biological membrane. The average orientation of the collagen bundles is used to guide where to cut the leaflet edges of the bioprosthetic heart valve.
The purpose of the invention is realized by the following technical scheme:
a biomembrane collagen beam orientation optical detection method based on weak coherent interference comprises the following steps:
imaging a biological membrane containing a collagen beam by OCT scanning to obtain an OCT biological membrane image;
performing transformation processing on the OCT biological membrane image at each depth to enable the surface of the biological membrane in the image to tend to be horizontal, and simultaneously keeping the height change of the collagen bundle;
and (4) selecting OCT biological membrane images with different depths to perform projection to obtain a projection image, and obtaining the average orientation of collagen beams with different depths according to the projection image.
The surface of one side of the biological membrane is smooth, and the surface of the other side of the biological membrane is provided with collagen bundles.
The biofilm tissue includes, but is not limited to, pericardium, aortic valve, dura mater, peritoneum, septum, or intestinal submucosa.
Imaging a biofilm containing collagen bundles using OCT scanning, comprising: and acquiring the interference spectrum of the biological membrane by utilizing OCT, extracting a depth domain intensity signal based on the interference spectrum of the biological membrane, and forming an OCT biological membrane image by the depth domain intensity signal.
Imaging a biofilm containing collagen bundles using OCT scanning, comprising:
a time domain OCT imaging method for changing the optical path of the reference arm by scanning;
or a spectral domain OCT imaging method for recording spectral interference signals by a spectrometer;
or a frequency-sweep OCT imaging method which utilizes a frequency-sweep light source to record spectral interference signals.
Transforming the coordinates of the OCT biomembrane image to make the biomembrane surface tend to be horizontal, and simultaneously keeping the height change of the collagen bundle, comprising the following steps:
air or moisture is arranged outside the surface of the biological membrane, the depth domain intensity signals of the pixel points are obviously different from the depth domain intensity signals of the surface of the biological membrane, and the difference of the depth domain intensity signals of the adjacent pixel points in the OCT biological membrane image is judged according to the difference between the surface of the biological membrane where the collagen bundles are located in the OCT biological membrane image and the depth domain intensity signals of the adjacent image area to obtain the surface boundary of the biological membrane where the collagen bundles are located; the specific implementation can set a difference threshold for judgment, and if the difference between the depth domain intensity signals of two adjacent pixels is greater than the difference threshold, the two adjacent pixels are regarded as a boundary.
Performing polynomial curve fitting on the surface boundary of the biological membrane to obtain a fitting result;
and transforming the depth coordinate of the biological membrane according to the fitting result to enable the surface of the biological membrane where the collagen bundle is located to be wholly horizontal, so that the height change of the collagen bundle is also kept.
Obtaining the average orientations of the collagen bundles at different depths according to the projection images, wherein the average orientations comprise: and performing fast Fourier transform on the projected image to obtain the oscillation frequency of the depth domain intensity signal of each pixel point in the collagen bundle image, and then calculating the orthogonal direction of the frequency distribution trend of the oscillation frequency to obtain the average orientation of the collagen bundle.
The orthogonal direction of the frequency distribution trend refers to the most main orthogonal direction in frequency distribution and can be obtained by adopting a clustering method.
Secondly, a biomembrane collagen beam orientation optical detection device based on weak coherent interference:
the OCT optical coherence detection device is used for collecting the interference spectrum of the biological membrane in a two-dimensional or three-dimensional space;
and one or more processors coupled to the OCT optical coherence detection apparatus for analyzing and processing the biomembrane interference spectrum to obtain the orientation of the biomembrane collagen beam.
The OCT optical coherence detection device adopts one of the following methods:
the system comprises a low-coherence light source, an interferometer and a detector;
or a low coherence light source, an interferometer and a spectrometer;
or a swept-bandwidth spectral light source, an interferometer and a detector.
According to the method, an optical coherence imaging system (OCT) is used for obtaining an interference spectrum of the biological membrane, Fourier transform is carried out on the interference spectrum to obtain a depth domain intensity signal, the surface coordinate of the biological membrane is calculated based on the depth domain intensity signal, then a polynomial fitting biological membrane surface curve is used, coordinate transform is carried out on the biological membrane according to the fitting curve to enable the surface of the biological membrane to tend to be horizontal, intensity signal projection of different depths is carried out on the flattened biological membrane, and the intensity signal projection graph can display the orientation characteristic of a collagen beam of the biological membrane.
The average orientation of the collagen bundles obtained by the present invention can be used to position where to cut the leaflet edges of a bioprosthetic heart valve.
Compared with the prior art, the invention has the following beneficial effects and advantages:
compared with polarized light measurement, the method can realize one-time imaging to obtain the biomembrane collagen bundle orientation with depth resolution (micrometer precision), and can realize rapid evaluation of cutting of the biomembrane edge to obtain high-quality artificial heart biological valve leaflets.
Drawings
FIG. 1 is a schematic diagram of the process of the present invention;
FIG. 2 is a schematic view of the apparatus of the present invention;
FIG. 3 is a schematic diagram of an embodiment of the present invention;
FIG. 4 is a diagram of the structure of the biomembrane collagen bundle OCT of the exemplary embodiment of the invention;
FIG. 5 is a drawing of biofilm collagen bundle orientation in accordance with an exemplary embodiment of the present invention.
Wherein: 1-collecting interference spectrum by using OCT; 2-extracting a depth domain intensity signal from the interference spectrum; 3, leveling processing is carried out based on the depth domain intensity signal; 4-obtaining the orientation of the biomembrane collagen bundles at different depths; 11-a light source; 12-a beam splitter; 13-reference arm collimator lens; 14-plane high reflector; 15-sample arm collimator; 16-a scanning galvanometer; 17-an objective lens; 18-a sample to be tested; 19-interference signal detection means; 20-a signal processor; 21-a polarization controller; 31-low coherence broadband light source; 32-an optical circulator; 33-a fiber coupler; 34-a first fiber alignment device; 35-a focusing lens; 36-plane high mirror; 37-a second fiber alignment device; 38-scanning galvanometer; 39-a focusing lens; 41-a third fiber collimating device; 42-a grating; a 43-Fourier transform lens; 44-high speed line camera; 45-signal processor module and computing unit; 46-a first polarization controller; 47-a second polarization controller; 48: a dispersion compensator; 49: a focusing lens; 50: a dichroic mirror; 51: a scanning lens.
Detailed Description
Reference will now be made in detail to the present embodiments of the invention, examples of which are illustrated in the accompanying drawings, which form a part hereof. It should be noted that the description and illustrations are exemplary only and should not be construed as limiting the scope of the invention, which is defined by the appended claims, as any variation based on the claims is intended to be within the scope of the invention.
To facilitate an understanding of embodiments of the invention, operations are described as multiple discrete operations, however, the order of description does not represent the order in which the operations are performed.
The x-y-z three-dimensional coordinate representation based on spatial direction is adopted for the sample measurement space in the description. This description is merely intended to facilitate discussion and is not intended to limit application of embodiments of the present invention. Wherein: the depth z direction is a direction along the incident optical axis; the x-y plane is a plane perpendicular to the optical axis, where x is orthogonal to y, and x denotes the OCT lateral fast scan direction and y denotes the slow scan direction.
The above I, x, y, z, etc. represent variables used merely to facilitate discussion and are not intended to limit the application of embodiments of the present invention, and can be any of values 1,2,3,4, etc.
As shown in figure 1, the method firstly utilizes OCT to collect interference spectrum, then extracts depth domain intensity signals from the interference spectrum, and then flattens the biological membrane based on the depth domain intensity signals to obtain collagen bundle images with different depths, and then calculates the average orientation of the collagen bundle images. And finally, selecting a target area in a proper direction to cut to obtain the biological valve leaflet of the artificial heart.
The method comprises the steps of collecting interference spectrum of a biological membrane tissue by using an OCT system, carrying out OCT scanning imaging of two-dimensional or three-dimensional space on the tissue sample, repeatedly scanning and imaging for a certain time at the same spatial position, and recording a spectrum interference signal by using a spectrometer (or a time domain OCT imaging method for changing the optical path of a reference arm by scanning and a sweep frequency OCT imaging method for recording the spectrum interference signal by using a sweep frequency light source).
And extracting a depth domain intensity signal by using the OCT interference spectrum, namely performing Fourier transform on the interference spectrum to obtain the depth domain intensity signal of each space point.
And extracting the surface boundary of the biological membrane where the collagen bundle is positioned based on the depth domain intensity signal. The method comprises the following specific steps: in the three-dimensional signal of the tissue of the biological membrane, I represents the OCT intensity signal, and the intensity difference dI (z, x, y) between the longitudinal coordinate y, the transverse coordinate x, and the adjacent depth coordinate z is calculated:
dI(z,x,y)＝I(z,x,y)-I(z-1,x,y) (1)
in the boundary position of the surface of the biological membrane, the difference between the intensity signals of the depth domain exists, so that the differential value dI is larger than that of other positions, so that the maximum value is selected by comparing dI (z, x, y1) of the same longitudinal coordinate y1, and the depth domain coordinate b (x) corresponding to the boundary is obtained. And performing polynomial fitting on the boundary coordinates b (x), and flattening the whole body but keeping the height change of the fiber bundle. The least square method is used for carrying out 15-order polynomial fitting, the fitting order can be adjusted according to the actual condition without limitation, and the fitted boundary coordinate c (x) is obtained. And then carrying out boundary flattening treatment on the original biomembrane intensity signal I (z, x, y) according to the fitted boundary coordinates c (x) to obtain a flattened biomembrane intensity signal I (z', x, y):
I(z′,x,y)＝I(z-c(x),x,y) (2)
and obtaining the biomembrane collagen bundle image I (x, y) under different depth coordinates z 'according to I (z', x, y), and performing Fourier transform on the biomembrane collagen bundle image I (x, y). The fourier transform allows to characterize the oscillation frequency of the intensity signal in the depth domain of each pixel point in the collagen bundle image, so that a frequency distribution related to the collagen bundle pitch can be obtained. And finally, extracting the direction of the collagen bundle orthogonal to the frequency distribution trend, namely the average orientation. And determining where to cut the leaflet edge of the biological valve of the artificial heart according to the obtained average orientation.
FIG. 2 is a schematic diagram of an optical detection apparatus for detecting the orientation of a collagen bundle of a biological membrane based on weak coherent interference according to the present invention. The main structure of the low coherence interferometry part of the device is an interferometer which is composed of 11-17, 19 and 21, wherein light emitted by a light source 11 is divided into two light beams by a beam splitter 12: one beam of light enters a reference arm of the interferometer and is irradiated on a plane high reflecting mirror 14 through a reference arm collimating mirror 13; the other beam of light enters the sample arm, is focused on a sample to be measured after being collimated by 15 and reflected by a light path; the sample 18 is placed at the focal plane of the sample arm objective 17. Then the light reflected back by the reference arm and the light reflected back by the sample arm are interfered and received by the interference signal detection device 19. For the optical fiber type optical path, the polarization controller 21 is adopted to adjust the polarization state of the light beam, and the signal interference effect is maximized.
According to different modes of low coherence interference detection signals, the device for optical detection of the orientation of the collagen bundle of the biological film based on weak coherence interference shown in fig. 2 specifically comprises:
1) a time domain measurement device. The light source 11 uses broadband low coherent light, the plane mirror 14 can move along the optical axis direction, and the interference signal detection device 19 is a point detector. The optical path of the reference arm is changed by moving the plane mirror 14, the interference signals of the two arms are detected by the point detector 19, and the low coherence interference detection is carried out on the scattered signals in the z direction of a certain space depth, so that a sampling body of the depth space dimension is obtained.
2) Spectral domain measuring device. The light source 11 adopts broadband low-coherence light, the plane reflector 14 is fixed, and the interference signal detection device 19 adopts a spectrometer. The interference signal passes through a linear array camera in the spectrometer and simultaneously records the interference spectrum. And analyzing the interference spectrum signals by adopting a Fourier analysis method, and parallelly acquiring scattering information in the depth z direction so as to obtain a sampling body of the depth dimension space.
3) Provided is a sweep frequency measuring device. The light source 11 adopts a sweep frequency light source, the plane reflector 14 is fixed, and the interference signal detection device 19 adopts a point detector. And the point detector records the low coherence interference spectrum of the swept-frequency light source in a time-sharing manner. And (3) carrying out Fourier analysis on the interference spectrum signal, and obtaining the scattering information in the depth z direction in parallel, thereby obtaining a sampling body of the depth dimension space.
For the different measuring devices, the orientation of the collagen bundle of the biological membrane can be analyzed respectively by combining the OCT scanning imaging mode related to the description of figure 1.
FIG. 3 illustrates one exemplary embodiment disclosed herein that utilizes the present invention. A biomembrane collagen beam orientation optical detection device based on weak coherent interference comprises a broadband low-coherence light source 31, an optical circulator 32, an optical fiber coupler 33 with a splitting ratio of 50:50, a first polarization controller 46, a first optical fiber collimating device 34, a focusing lens 35, a plane high-reflection mirror 36, a second polarization controller 47, a second optical fiber collimating device 37, a scanning galvanometer 38, an objective lens 39, a sample dispersing device 40, a third optical fiber collimating device 41, a grating 42, a Fourier transform lens 43, a high-speed line camera 44, a signal processor module and calculation unit 45, a color compensator 48, a focusing lens 49, a dichroic mirror 50, a scanning lens 51, a die cutting assembly 100, and displacement tables 102 and 104 with optical transparent plates: vertical displacement device 104, biofilm tissue 106, transparent glass table 108, mounting bar 114, connecting arm 115, height display 118, mold 120. The broadband low-coherence light source 31 adopts a super-light-emitting diode light source with the central wavelength of 1325nm and the bandwidth of 100nm, the high-speed linear array camera 44 adopts a linear array scanning camera consisting of 2048 pixel units, and the scanning lens 51 in the sample arm adopts a lens with the focal length of 54 mm.
The light emitted by the low coherence broadband light source 31 used by the device of the present invention enters the optical fiber coupler 33 with a splitting ratio of 50:50 after passing through the optical circulator 32, and the light emitted from the optical fiber coupler 33 is divided into two sub-beams: one beam of light is connected to a first optical fiber collimating device 34 in the reference arm through an optical fiber through a first polarization controller 46, and is irradiated to a plane high reflecting mirror 36 after being subjected to collimation, dispersion compensation of a dispersion compensator 48 and focusing of a focusing lens 35; the other beam of light is connected to a second optical fiber collimating device 37 of the sample arm part through an optical fiber via a second polarization controller 47, and is irradiated on the measured sample after being reflected by the light path of the collimating and scanning galvanometer 38 and focused by the focusing lens 39 and the focusing lens 49. Before the scanning lens 51, the dichroic mirror 50 is used to make the 90 ° turn of the OCT probe light path by the light emitted from the focusing lens 49. The scanning galvanometer 38 in the sample arm is fixed, so that the low coherence interferometer can parallelly detect and obtain the scattering signals of the same position in the sample space in the depth direction at different moments. Meanwhile, a light path in the sample arm conducts light beams through the single-mode optical fiber, and the light scattered back by the sample to be detected plays a role in spatial filtering, namely multiple scattering components in scattering signals are effectively reduced. The light reflected by the plane high reflecting mirror 36 in the reference arm interferes with the light backscattered from the sample to be measured in the sample arm at the optical fiber coupler 33, the interference light is detected and recorded by a spectrometer (comprising devices 41-44), and then the interference light is collected by a computing unit 45 and is subjected to signal analysis processing.
According to the invention, the orientation of the biological membrane fiber bundle can be obtained according to the OCT depth domain intensity signal, and the method can be used for guiding the cutting of the leaflet edge of the artificial heart biological valve and improving the strength of the manufactured leaflet.
Fig. 4 shows the imaging result of the collagen bundle on the surface layer of the flattened biological membrane, which is a structural diagram for OCT acquisition, and OCT can realize three-dimensional structural imaging. Fig. 4a is a projection view of the structure of the collagen beam OCT, and fig. 4b is a cross-sectional view of the dotted line in fig. 4 a. Fig. 4c is a projection view of the OCT collagen bundle structure, and fig. 4d is a cross-sectional view of the dotted line in fig. 4 c. Images of collagen bundles at any depth can be obtained, and fig. 4b and 4d correspond to the flattened biofilm and the flattened biofilm respectively, so that the overall level of the surface of the biofilm can be seen, but the height change of the collagen bundles on the surface is retained.
FIG. 5 shows the results of the average orientation of collagen bundles in the biofilm. Fig. 5a is a projection view of the OCT structure of the collagen bundle, and fig. 5b is a result of the average orientation of the collagen bundle after the fast fourier transform, the average orientation being indicated by a black solid line. The calculated average orientation corresponds to the characteristic of the collagen bundle in the projection.
The above experimental results fully illustrate that: the invention can realize nondestructive acquisition of the orientation of the collagen bundles of the biological membrane at different depths and realize rapid evaluation of the cutting direction of the biological membrane so as to acquire the leaflet of the artificial heart biological valve.
Claims (8)
1. A biomembrane collagen beam orientation optical detection method based on weak coherent interference is characterized by comprising the following steps:
imaging a biological membrane containing a collagen beam by OCT scanning to obtain an OCT biological membrane image;
performing transformation processing on the OCT biological membrane image at each depth to enable the surface of the biological membrane in the image to tend to be horizontal, and simultaneously keeping the height change of the collagen bundle;
and (4) selecting OCT biological membrane images with different depths to perform projection to obtain a projection image, and obtaining the average orientation of collagen beams with different depths according to the projection image.
2. The method for optical detection of the orientation of the collagen bundle in the biological membrane based on the weak coherent interference as claimed in claim 1, wherein: the biofilm tissue comprises pericardium, aortic valve, dura mater, peritoneum, septum, or intestinal submucosa.
3. The method for optical detection of the orientation of the collagen bundle in the biological membrane based on the weak coherent interference as claimed in claim 1, wherein: imaging a biofilm containing collagen bundles using OCT scanning, comprising: and acquiring the interference spectrum of the biological membrane by utilizing OCT, extracting a depth domain intensity signal based on the interference spectrum of the biological membrane, and forming an OCT biological membrane image by the depth domain intensity signal.
4. The method for optical detection of the orientation of the collagen bundle in the biological membrane based on the weak coherent interference as claimed in claim 1, wherein: imaging a biofilm containing collagen bundles using OCT scanning, comprising:
a time domain OCT imaging method for changing the optical path of the reference arm by scanning;
or a spectral domain OCT imaging method for recording spectral interference signals by a spectrometer;
or a frequency-sweep OCT imaging method which utilizes a frequency-sweep light source to record spectral interference signals.
5. The method for optical detection of the orientation of the collagen bundle in the biological membrane based on the weak coherent interference as claimed in claim 1, wherein: transforming the coordinates of the OCT biomembrane image to make the biomembrane surface tend to be horizontal, and simultaneously keeping the height change of the collagen bundle, comprising the following steps:
according to the difference between the surface of the biological membrane where the collagen bundle in the OCT biological membrane image is located and the depth domain intensity signal of the adjacent image area, carrying out difference judgment on the depth domain intensity signal of the adjacent pixel point in the OCT biological membrane image to obtain the surface boundary of the biological membrane where the collagen bundle is located;
performing polynomial curve fitting on the surface boundary of the biological membrane to obtain a fitting result;
and transforming the depth coordinate of the biological membrane according to the fitting result to enable the surface of the biological membrane where the collagen bundle is located to be wholly horizontal, so that the height change of the collagen bundle is also kept.
6. The method for optical detection of the orientation of the collagen bundle in the biological membrane based on the weak coherent interference as claimed in claim 1, wherein: obtaining the average orientations of the collagen bundles at different depths according to the projection images, wherein the average orientations comprise: and performing fast Fourier transform on the projected image to obtain the oscillation frequency of the depth domain intensity signal of each pixel point in the collagen bundle image, and then calculating the orthogonal direction of the frequency distribution trend of the oscillation frequency to obtain the average orientation of the collagen bundle.
7. An optical detection device for detecting the orientation of a collagen bundle of a biological film based on weak coherent interference, which is used for implementing the method of any one of claims 1 to 6, and is characterized by comprising:
the OCT optical coherence detection device is used for collecting the interference spectrum of the biological membrane in a two-dimensional or three-dimensional space;
and one or more processors coupled to the OCT optical coherence detection apparatus for analyzing and processing the biomembrane interference spectrum to obtain the orientation of the biomembrane collagen beam.
8. The apparatus according to claim 7, wherein the optical detection device for detecting the orientation of the collagen bundle in the biological membrane based on weak coherent interference comprises: the OCT optical coherence detection device adopts one of the following methods:
the system comprises a low-coherence light source, an interferometer and a detector;
or a low coherence light source, an interferometer and a spectrometer;
or a swept-bandwidth spectral light source, an interferometer and a detector.
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