Noninvasive detection method and noninvasive detection device for circulating melanoma cells

Abstract

The invention discloses a noninvasive detection method and a noninvasive detection device for circulating melanoma cells, and relates to the technical field of noninvasive detection of melanoma. A noninvasive detection method for circulating melanoma cells comprises the following steps that a first light beam and a second light beam which are of the same light source and have different wavelengths sequentially irradiate a position area of a target blood vessel in a pulse mode, and after the target blood vessel is irradiated, a first photoacoustic signal and a second photoacoustic signal are sequentially generated; the signal intensity of the first photoacoustic signal is used for feeding back the coverage condition of irradiation of the first light beam on the target blood vessel; the signal intensity of the second photoacoustic signal is used for representing clinical index information of melanoma in the target blood vessel. According to the clinical noninvasive detection method for melanoma designed by the invention, rapid and accurate noninvasive detection of melanoma can be realized; the built noninvasive detection device for melanoma is simple in building process, and the built noninvasive detection device is relatively high in light path stability and relatively high in anti-interference capability.

Description

[0001] This invention claims priority from application number 2023111429185 Technical Field

[0002] The present invention relates to the technical field of non-invasive detection of melanoma, and in particular to a non-invasive detection method and a non-invasive detection device for circulating melanoma cells. Background Art

[0003] Melanoma is a highly metastatic malignant tumor. Melanoma cells that shed from the tumor in situ and enter the circulatory system are called melanoma circulating tumor cells (CTCs), which are important markers of tumor metastasis. The detection of melanoma CTCs, especially real-time and non-invasive detection, has always been an urgent clinical need.

[0004] The photoacoustic effect is the result of the conversion between light and sound waves caused by the light absorption and local thermal expansion of the material when a pulsed laser is irradiated onto a biological or tissue sample. Cells can be detected by detecting ultrasonic signals. The photoacoustic effect is used in more and more fields as an important tool in spectral analysis. Whether the photoacoustic effect can be achieved is related to the light absorption coefficient of the material.

[0005] Studies have found that the light absorption coefficient of melanin at a wavelength of 1064nm is significantly different from that of other substances, that is, photoacoustics are particularly sensitive to the detection of melanin. Therefore, label-free in vivo flow cytometry based on the photoacoustic effect is an ideal method for non-invasive detection of melanoma cells.

[0006] At the same time, due to the differences in the blood vessels of the subjects during the non-invasive detection of melanoma cells by label-free in vivo flow cytometry based on the photoacoustic effect, when the pulsed laser with a wavelength of 1064nm is used to irradiate and detect the blood vessels of the subjects, it is often unable to quickly and accurately locate the maximum coverage cross-section of the blood vessels to be tested, thereby affecting the accuracy of the detection.

[0007] Therefore, how to design a non-invasive detection method for circulating melanoma cells, which can quickly locate the position of the blood vessels to be detected and accurately detect melanoma, is a technical problem that technicians in this field urgently need to solve.

[0008] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0009] In view of the above technical problems, the embodiments of the present invention provide a non-invasive detection method and a non-invasive detection device for circulating melanoma cells to solve the problems raised in the above background technology.

[0010] The present invention provides the following technical solutions:

[0011] A method for non-invasive detection of circulating melanoma cells comprising the following steps:

[0012] A first light beam and a second light beam with different wavelengths from the same light source are sequentially irradiated in a pulsed manner to the target blood vessel, and the target blood vessel generates a first photoacoustic signal and a second photoacoustic signal sequentially after being irradiated;

[0013] The signal strength of the first photoacoustic signal is used to feedback the coverage of the first light beam on the target blood vessel; the signal strength of the second photoacoustic signal is used to characterize the clinical indicator information of melanoma in the target blood vessel.

[0014] As a further improvement of the present invention, the wavelength of the first light beam is 532 nm; after the first light beam is irradiated on the location area of ​​the target blood vessel in a pulsed manner, the hemoglobin flowing through the target blood vessel will generate a first photoacoustic signal.

[0015] As a further improvement of the present invention, the wavelength of the second light beam is 1064 nm; after the second light beam is irradiated on the location area of ​​the target blood vessel in a pulsed manner, the melanoma cells flowing through the target blood vessel will generate a second photoacoustic signal.

[0016] As a further improvement of the present invention, the first photoacoustic signal and the second photoacoustic signal are collected using an ultrasonic probe.

[0017] As a further improvement of the present invention, in the step where the signal strength of the first photoacoustic signal is used to feedback the coverage of the first light beam irradiated on the target blood vessel, the position of the first light beam irradiated on the target blood vessel can be adjusted according to the signal strength of the first photoacoustic signal until the signal strength value of the first photoacoustic signal is higher than the threshold value of the first photoacoustic signal;

[0018] The signal intensity of the second photoacoustic signal is used in the step of characterizing clinical indicator information of melanoma in the target blood vessel, wherein the clinical indicator information includes the number of melanoma cells and the size and number of melanoma cell clusters.

[0019] A detection device for the non-invasive detection method of circulating melanoma cells as described above comprises: an optical excitation module, a photoacoustic signal receiving module and a signal data processing module;

[0020] The optical excitation module is used to emit a first light beam and a second light beam with different wavelengths; the first light beam and the second light beam irradiate the location area of ​​the target blood vessel in a pulsed manner and generate a first photoacoustic signal and a second photoacoustic signal;

[0021] The signal strength of the first photoacoustic signal is used to feedback the coverage area of ​​the first light beam irradiated on the target blood vessel; the signal strength of the second photoacoustic signal is used to characterize the clinical indicator information of melanoma in the target blood vessel;

[0022] The photoacoustic signal receiving module is used to receive the first photoacoustic signal and the second photoacoustic signal, and convert the received first photoacoustic signal and the second photoacoustic signal into corresponding electrical signals and amplify them;

[0023] The signal data processing module is used to process and store the received electrical signals.

[0024] As a further improvement of the present invention, the optical excitation module includes: a laser light source, a first mechanical slit, a cylindrical lens, a convex lens, a second mechanical slit, a microscope objective lens and a signal generator;

[0025] The laser light source, the first mechanical slit, the cylindrical lens, the convex lens, the second mechanical slit and the microscope objective lens are coaxially installed in the optical path in sequence; the laser light source and the signal generator are electrically connected.

[0026] As a further improvement of the present invention, it also includes a light beam direction adjustment unit; the light beam direction adjustment unit includes a plurality of reflectors; the plurality of reflectors are respectively coaxially inserted in the light path of the optical excitation module.

[0027] As a further improvement of the present invention, the photoacoustic signal receiving module includes an ultrasonic probe, a signal amplifier, and a pulse generator which are electrically connected.

[0028] As a further improvement of the present invention, the signal data processing module includes a host computer and a data acquisition card; the host computer processes the received data, and the data acquisition card stores the processed data.

[0029] A non-invasive detection method and a non-invasive detection device for circulating melanoma cells provided by an embodiment of the present invention have the following beneficial effects: the present invention designs a clinical non-invasive detection method for melanoma, which can quickly locate the maximum coverage position of the blood vessel to be tested, thereby realizing rapid and accurate non-invasive detection of melanoma; the present invention also constructs a non-invasive detection device for melanoma, the construction process of the non-invasive detection device is simple and convenient, and the constructed non-invasive detection device has high optical path stability and strong anti-interference ability. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a schematic diagram of the structure of the detection device for non-invasive detection of melanoma in the present invention. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0032] In view of the problems mentioned in the above background technology, the embodiments of the present invention provide a non-invasive detection method and a non-invasive detection device for circulating melanoma cells to solve the above technical problems. The technical solutions are as follows:

[0033] Example 1

[0034] A method for non-invasive detection of circulating melanoma cells comprising the following steps:

[0035] A first light beam and a second light beam with different wavelengths from the same light source are sequentially irradiated in a pulsed manner to the target blood vessel, and the target blood vessel generates a first photoacoustic signal and a second photoacoustic signal after being irradiated; the target blood vessel is a blood vessel to be measured in the patient;

[0036] The signal strength of the first photoacoustic signal is used to feedback the coverage of the first light beam on the target blood vessel; the signal strength of the second photoacoustic signal is used to characterize the clinical indicator information of melanoma in the target blood vessel.

[0037] A detection device for non-invasive detection of melanoma: comprising: an optical excitation module, a photoacoustic signal receiving module and a signal data processing module;

[0038] The optical excitation module is used to emit a first light beam and a second light beam with different wavelengths; the first light beam and the second light beam irradiate the location area of ​​the target blood vessel in a pulsed manner and generate a first photoacoustic signal and a second photoacoustic signal;

[0039] The signal strength of the first photoacoustic signal is used to feedback the coverage area of ​​the first light beam irradiated on the target blood vessel;

[0040] The signal intensity of the second photoacoustic signal is used to characterize the clinical indicator information of melanoma in the target blood vessel; the clinical indicator information includes the number of melanoma cells and the size and number of melanoma cell clusters;

[0041] The photoacoustic signal receiving module is used to receive the first photoacoustic signal and the second photoacoustic signal, and convert the received first photoacoustic signal and the second photoacoustic signal into corresponding electrical signals and amplify them respectively; the first photoacoustic signal and the second photoacoustic signal are collected by an ultrasonic probe; the ultrasonic probe is located near the focused strip-shaped laser spot;

[0042] The signal data processing module is used to process and store the received electrical signals.

[0043] Preferably, the wavelength of the first light beam is 532nm; the light beam with a wavelength of 532nm is green light, which can play a role in blood vessel navigation; since hemoglobin in the background components of blood vessels has a strong absorption of green light, an obvious photoacoustic signal is generated; based on the signal intensity generated by green light excitation, it can be determined whether the light beam is positioned and covers the optimal detection position in the blood vessel, that is, the maximum coverage cross-section.

[0044] Preferably, the wavelength of the second light beam is 1064nm; the light beam with a wavelength of 1064nm is near-infrared light; the light absorption coefficient of melanin in melanoma at a wavelength of 1064nm is significantly different from that of other substances, that is, photoacoustics are particularly sensitive to the detection of melanin; after the green navigation light with a wavelength of 532nm determines the optimal detection position, switching to near-infrared can ensure that the excitation light is focused on the area through which the CTC flows, thereby making the detected melanoma CTC signal as strong as possible and improving the system signal-to-noise ratio.

[0045] Preferably, the position at which the first light beam irradiates the target blood vessel can be adjusted according to the signal strength of the first photoacoustic signal until the first light beam is modulated to irradiate the target blood vessel to obtain a maximum coverage cross section; the maximum coverage cross section is determined by comparing the signal strength value of the first photoacoustic signal collected in real time with the threshold of the first photoacoustic signal until the signal strength value of the collected first photoacoustic signal is higher than the threshold of the first photoacoustic signal, indicating that the position at which the first light beam irradiates the target blood vessel can obtain the maximum coverage cross section.

[0046] Preferably, the optical excitation module comprises: a laser light source 1, a first mechanical slit 8, a cylindrical lens 2, a convex lens 3, a second mechanical slit 9, a microscope objective lens 10 and a signal generator;

[0047] The laser light source 1, the first mechanical slit 8, the cylindrical lens 2, the convex lens 3, the second mechanical slit 9, and the microscope objective lens 10 are coaxially installed in the optical path in sequence; the laser light source 1 is electrically connected to the signal generator.

[0048] The light beam emitted by the laser light source 1 passes through the first mechanical slit 8 and is reflected into the cylindrical lens 2. The laser forms a stripe light spot after passing through the cylindrical lens 2, and then is emitted to the objective lens 10 through the adjustment of the convex lens 3 and the second mechanical slit 9 to converge into an elliptical Gaussian light spot, thus serving as an excitation source. The short axis of the light spot is about 20μm and the long axis is about 1000μm.

[0049] Preferably, it also includes a light beam direction adjustment unit; the light beam direction adjustment unit includes multiple reflectors; the multiple reflectors are coaxially inserted in the optical path of the optical excitation module; wherein the purpose of arranging the light beam direction adjustment unit in the optical path of the optical excitation module is to modulate or change the light beam direction to achieve miniaturization of the equipment.

[0050] Preferably, the beam direction adjustment unit includes a first reflector 4, a second reflector 5, a third reflector 11, a fourth reflector 12, a fifth reflector 6 and a sixth reflector 7; the first reflector 4, the second reflector 5, the third reflector 11, the fourth reflector 12, the fifth reflector 6 are arranged in sequence between the laser light source 1 and the first mechanical slit 8; the sixth reflector 7 is arranged between the second mechanical slit 9 and the objective lens 10.

[0051] Preferably, a sample stage 13 is further included. The sample stage 13 is located below the objective lens 10 and is used to place the patient's arm.

[0052] Preferably, the photoacoustic signal receiving module includes an electrically connected ultrasound probe, a signal amplifier, and a pulse generator.

[0053] Preferably, the signal data processing module includes a host computer and a data acquisition card; the host computer processes the received data, and the data acquisition card stores the processed data.

[0054] It should be noted that after completing the signal acquisition, the data is generally processed by using a baseline correction algorithm and a related deep neural network to perform signal noise reduction, screen the pre-processed signals, set the screening threshold according to the cell size, and continuously appear on the time axis. Signals with a duration higher than the threshold are recorded as CTC clusters, and signals below the threshold are recorded as single CTCs.

[0055] Example 2

[0056] A specific operation method of a detection device for non-invasive detection of melanoma:

[0057] 1. Select the patient's blood vessel to be tested: the patient makes a fist with his right hand, observes on the back of his hand, and selects a blood vessel with a clear outline and prominent color; apply ultrasonic coupling agent on this blood vessel; the coupling agent is used to transmit the ultrasonic signal to the ultrasonic probe;

[0058] 2. Turn on the detection device: turn on each electrical component on the detection device in turn, and adjust the working parameters of each electrical component;

[0059] 3. Fine-tuning of the photoacoustic position of the detection device: During the measurement process, it is necessary to ensure that the focus of the light excitation is within the range of the acoustic detection. Use the laser line color card to determine the focus position after the objective lens emits light, so that the position of the ultrasonic probe coincides with the light focus, and the position of the ultrasonic probe is placed 2-3mm above the light focus;

[0060] After the laser emits light, it passes through a cylindrical mirror and a plano-convex lens to compress the light spot into a strip-shaped light spot, and finally emits it from the objective lens; wherein, the direction of the light can be adjusted in the optical path with the help of a beam direction adjustment unit;

[0061] 4. Patient test: The patient's hand to be tested is straightened and placed on the height adjustment stand. The height and hand position are adjusted so that the ultrasound probe is immersed in / fitted into the coupling agent.

[0062] 5. Measurement plan:

[0063] a. Use 532nm laser to observe the signal of the blood vessel to be tested, and record 12 seconds of data after the signal stabilizes;

[0064] b. Then switch to 1064nm laser to observe the signal of the blood vessel to be tested, and record 120 seconds of data after the signal stabilizes;

[0065] c. Then switch to 532nm laser to confirm that the blood vessel signal is stable;

[0066] d. Finally, switch to 1064nm laser and record 120 seconds of data after the signal stabilizes.

[0067] 6. After data collection is completed, lower the height adjustment frame to separate the coupling agent on the patient's hand from the acoustic probe, and then the patient withdraws his hand.

[0068] 7. Use data processing programs to identify the number and clusters of melanoma CTCs cells measured in the patient.

[0069] Example 3

[0070] 1. Use a laser to generate dual-wavelength laser to synchronously excite melanoma CTCs in the circulating system through the transformation, shaping and focusing of the optical path.

[0071] 2. In this embodiment, the wavelengths of the dual-wavelength laser are 532nm and 1064nm respectively, the pulse width of the excitation light source is tp≤50-500ps, and the pulse frequency of the excitation light source is determined according to the diameter of the blood vessel to be measured, the width of the excitation light, and the volume of melanoma CTCs. The selection of laser parameters must be lower than the MPE (maximum permissible exposure) value before it can be used clinically;

[0072] The transformation, shaping and focusing of the optical path are achieved by compressing the shape of the light spot by adjusting the position of the convex lens and the concave lens, compressing the light spot into a strip-shaped spot, and the adjustable aperture is used to remove stray light, so that the laser light spot is shaped into a strip-shaped spot and focused, forming a laser window to cover the blood vessel to be measured.

[0073] Through software design and pulse generator, two wavelengths of laser are synchronously irradiated to tumor cells in the circulatory system to generate photoacoustic signals.

[0074] 4. The photoacoustic signal is collected by an ultrasonic probe, amplified by a signal amplifier, and then collected, recorded and stored by a high-speed acquisition card.

[0075] 5. In this embodiment, the ultrasonic probe adopts a non-focused ultrasonic probe, and the bandwidth of the amplification circuit is 3-5 times the center frequency of the received signal.

[0076] 6. The high-speed acquisition card is connected to the computer and transmits the data to the computer into the signal processing system.

[0077] The device embodiments described above are merely illustrative, wherein the units described as separate components may or may not be physically separated, and the components shown as units may or may not be physical units, and may be located in one place or distributed over multiple network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0078] In the present invention, unless otherwise clearly stipulated and limited, the terms such as "installation", "setting", "connection", "fixation" and "screw-on" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integrated connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, and it can be the internal connection of the components or the interaction relationship between the components. Unless otherwise clearly defined, for ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to the specific circumstances.

[0079] It is understandable that those skilled in the art can make equivalent substitutions or changes based on the technical solution and concept of the present invention, and all these changes or substitutions should fall within the protection scope of the claims attached to the present invention.

Claims

1. A method for non-invasive detection of circulating melanoma cells, characterized in that: The following steps are involved: A first light beam and a second light beam with different wavelengths from the same light source are sequentially irradiated in a pulsed manner to the target blood vessel, and the target blood vessel generates a first photoacoustic signal and a second photoacoustic signal sequentially after being irradiated; The signal strength of the first photoacoustic signal is used to feedback the coverage of the first light beam on the target blood vessel; the signal strength of the second photoacoustic signal is used to characterize the clinical indicator information of melanoma in the target blood vessel.

2. The non-invasive detection method for circulating melanoma cells according to claim 1, characterized in that The wavelength of the first light beam is 532 nm. After the first light beam is irradiated on the location area of ​​the target blood vessel in a pulsed manner, the hemoglobin flowing through the target blood vessel will generate a first photoacoustic signal.

3. The non-invasive detection method for circulating melanoma cells according to claim 1, characterized in that: The wavelength of the second light beam is 1064 nm. After the second light beam is irradiated on the location area of ​​the target blood vessel in a pulsed manner, the melanoma cells flowing through the target blood vessel will generate a second photoacoustic signal.

4. The non-invasive detection method for circulating melanoma cells according to claim 1, characterized in that: The first photoacoustic signal and the second photoacoustic signal are collected by using an ultrasonic probe.

5. The non-invasive detection method for circulating melanoma cells according to claim 1, characterized in that: In the step where the signal strength of the first photoacoustic signal is used to provide feedback on the coverage of the first light beam irradiated on the target blood vessel, the position of the first light beam irradiated on the target blood vessel can be adjusted according to the signal strength of the first photoacoustic signal until the signal strength value of the first photoacoustic signal is higher than the threshold value of the first photoacoustic signal; The signal intensity of the second photoacoustic signal is used in the step of characterizing clinical indicator information of melanoma in the target blood vessel, wherein the clinical indicator information includes the number of melanoma cells and the size and number of melanoma cell clusters.

6. A detection device for the non-invasive detection method of circulating melanoma cells as claimed in claim 1, characterized in that: include: Optical excitation module, photoacoustic signal receiving module and signal data processing module; The optical excitation module is used to emit a first light beam and a second light beam with different wavelengths; the first light beam and the second light beam irradiate the location area of ​​the target blood vessel in a pulsed manner and generate a first photoacoustic signal and a second photoacoustic signal; The signal strength of the first photoacoustic signal is used to feedback the coverage area of ​​the first light beam irradiated on the target blood vessel; the signal strength of the second photoacoustic signal is used to characterize the clinical indicator information of melanoma in the target blood vessel; The photoacoustic signal receiving module is used to receive the first photoacoustic signal and the second photoacoustic signal, and convert the received first photoacoustic signal and the second photoacoustic signal into corresponding electrical signals and amplify them; The signal data processing module is used to process and store the received electrical signals.

7. The detection device for the non-invasive detection method of circulating melanoma cells according to claim 6, characterized in that: The optical excitation module includes: a laser light source, a first mechanical slit, a cylindrical lens, a convex lens, a second mechanical slit, a microscope objective lens and a signal generator; The laser light source, the first mechanical slit, the cylindrical lens, the convex lens, the second mechanical slit and the microscope objective lens are coaxially installed in the optical path in sequence; the laser light source and the signal generator are electrically connected.

8. The detection device for the non-invasive detection method of circulating melanoma cells according to claim 7, characterized in that: It also includes a light beam direction adjustment unit; the light beam direction adjustment unit includes a plurality of reflectors; the plurality of reflectors are coaxially inserted in the light path of the optical excitation module.

9. The non-invasive detection method for circulating melanoma cells according to claim 6, characterized in that: The photoacoustic signal receiving module includes an ultrasonic probe, a signal amplifier, and a pulse generator which are electrically connected.

10. The non-invasive detection method for circulating melanoma cells according to claim 6, characterized in that: The signal data processing module includes a host computer and a data acquisition card; the host computer processes the received data, and the data acquisition card stores the processed data.

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