KR20170097426A - Label-free Imaging System for Specific Detection of Peripheral Nerve - Google Patents
Label-free Imaging System for Specific Detection of Peripheral Nerve Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/40—Detecting, measuring or recording for evaluating the nervous system
- A61B5/4029—Detecting, measuring or recording for evaluating the nervous system for evaluating the peripheral nervous systems
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0033—Features or image-related aspects of imaging apparatus classified in A61B5/00, e.g. for MRI, optical tomography or impedance tomography apparatus; arrangements of imaging apparatus in a room
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0077—Devices for viewing the surface of the body, e.g. camera, magnifying lens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0082—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes
Abstract
Description
The present disclosure relates to systems and methods for selectively imaging peripheral nerves in vivo without the use of external markers. More specifically, the present disclosure is based on the thin film interference phenomenon arising from the unique lamellar structure of the nerve fiber bundle, and it is possible to use strong reflected light emitted from the peripheral nerve at a specific excitation wavelength, To a system and method for selectively imaging peripheral nerves relative to tissue.
A variety of medical techniques suitable for imaging biological tissue and organs are known. Examples of such techniques are typically x-rays, ultrasound, magnetic resonance imaging, computerized tomographs (CT), and the like.
In recent years, there has been a growing demand for imaging techniques for nerve tissues including peripheral nerves. For example, these peripheral nerves may be difficult to distinguish from peripheral muscles and adipose tissues during surgery, In addition, guidance for local anesthesia or imaging for peripheral nerves is required for the diagnosis of neuropathy, which is often caused by diabetic complications, and for guidance in the treatment of neuropathy.
In general, the axons of peripheral nervous system neurons are surrounded by schwann cells and may be surrounded by oligodendrocytes in neurons of the central nervous system. The cell membranes of Schwann cells and rarely proliferating glial cells are specifically differentiated to surround the axons in multiple layers and fuse together. The cell membrane structure of the fused Schwann cells surrounding the axon is called myelin sheath, and the myelin component is called the myelin. Neurons that make up most of the peripheral nerves, such as the motor nerves and the sensory nerves, are surrounded by aqueducts, while others are not.
The axons surrounded by aqueducts are named as myelinated axons or myelinated nerve fibers while the axons not surrounded by aqueducts are called unmyelinated axons or anhydrous nerve fibers unmyelinated nerve fiber). At this time, a few seconds is a complex cell structure that plays an important role in propagation, axonal insulation, and trophic support. While the axon is mainly composed of water, the myelin plant is composed of 80% lipid and 20% protein.
In the field of peripheral nerve imaging as described above, in vivo and intraoperative imaging techniques are essential.
For example, there is a problem in that when the surgical operation can not accurately distinguish the nervous tissue from the other tissues, side effects such as cutting or damage of the nerve may be caused.
In addition, since a peripheral nerve blocking process is required in local anesthesia, it is required to position the needle and local anesthetic next to the target nerve. Thus, the success of blocking on the peripheral nerve is mainly determined by the position of the needle tip and the subsequent position of the agent to be administered. Thus, a reliable neural localization technique is required in determining the precise position for local anesthesia, thereby accurately and securely positioning the needle in a position immediately adjacent to the peripheral nerve.
As described above, precisely grasping the position of the peripheral nerve is indispensable for various surgical operations, local anesthesia, neuropathic diagnosis, and nerve stimulation therapy.
In this regard, current research on neural tissue imaging techniques is under way, for example, (1) electrical stimulation (see, for example, Korean Patent Publication No. 2009-0112728, Korean Patent No. 1270935 (2) Ultrasound, (3) MRI, (4) Fluorescent probe (for example, Michael A Whitney et al., Fluorescent peptides highlight peripheral nerves during surgery in mice, Nature Biotechnology, Vol. 29, No. 4 (APRIL 2011)), (5) Third Harmonic Generation microscopy, (6) Optical coherence tomography, (7) Raman microscopy ), (8) photoacoustic tomography, and (9) optical spectroscopy.
In the case of techniques using fluorescent probes (or labels), tricarbosilane dyes such as indocyanine green (ICG) have been used as nonspecific fluorescent probes in traditional medical imaging applications. Alternatively, fluorescent dyes such as infracian green, prororesin isothiocyanate, and rhodamine are used. Fluorescent probes detect light of an emission wavelength as a substance or component (including a small molecule (dye), protein, other polymer or macromolecule) that emits light of a longer wavelength when excited by energy of a specific wavelength, It is possible to track the position and concentration of the probe (U.S. Patent Publication No. 2014/0276008). Specifically, the fluorescent probe-based surgical guide technique increases the visual contrast effect on specific tissues and organs during surgery, thereby providing a more effective visual discrimination performance between the tissue to be cut and the tissue to be protected as compared with the effect of white light alone. For this purpose, techniques using confocal and two-photon microscopy using fluorescent labels are known (Kirby BB et al ., In vivo time-lapse imaging shows dynamic oligodendrocyte progenitor behavior during zebrafish development. Nat Neurosci. 2006; 9: 1506- 11). However, it does not provide a technique for selectively imaging peripheral nerves differently from major biological tissues such as blood vessels, muscles, and fats. Fluorescent dyes such as fluoromyelin among commercially available products may be useful for biological imaging, It is not allowed. Recently, a number of peripheral nerve-specific fluorescent probes based on peptide and synthetic dyes have been developed and reported, but there are no candidates for clinical trials (Michael A Whitney et al ., Fluorescent peptides highlight peripheral nerves during surgery in mice, Nature Biotechnology, Vol. 29, No. 4 (APRIL 2011)). Most of all, the toxicity of fluorescent probes is the most problematic. In reality, safety is recognized as a fluorescent contrast agent, and the types approved by the FDA and the KFDA are limited, and there is no probe that selectively stains peripheral nerves.
In the case of optical coherence tomography (OCT), the principle of low-coherence interferometry or white-light interferometry and the principle of confocal microscopy are combined to image the microstructure inside the biotissue, Basically, a Michelson interferometer using a light source with a very short coherence is used. Structural and morphological observations using OCT can also visualize peripheral nerves, but do not show peripheral nerve-specific contrast compared to other tissues.
In addition, the electric stimulation method, which is mainly used in the current clinical field, is a point simulation. Since the threshold value depends on the distance, it can be applied only to a narrow region. Since the contrast ratio is low, Contrast peripheral nerves are difficult to distinguish. With the Raman microscope, the device setup is complex and relatively expensive.
In addition, other imaging methods that do not utilize external markers in the industry have great potential, but require complicated device setup and facilities that are not easy to obtain, and that it is difficult to perform real-time imaging due to complicated post- And the like.
Therefore, there is still a need for a technique capable of selectively real-time biometric imaging of the location of peripheral nerves without using external marking techniques such as fluorescence probes, while solving the limitations of the prior art described above.
SUMMARY OF THE INVENTION The present disclosure provides a system and imaging method that can selectively and realistically image the location of peripheral nerves in the human body without the use of conventionally known biomedical techniques, particularly, external labeling materials such as fluorescent probes.
According to a first aspect of the present disclosure,
A system for selectively imaging peripheral nerves in a subject,
A light source arranged to illuminate the object to be inspected, wherein the light source comprises a first light source providing a beam of at least 600 nm wavelength band and a second light source providing a beam of visible light band, Wherein the first light source and the second light source are configured to alternately illuminate an inspection target;
Each of the beams alternately irradiated from the first light source and the second light source transmits a first light component indicative of a peripheral nerve among reflected light emitted from the object to be inspected through a peripheral nerve membrane phenomenon of the peripheral nerve, A beam splitter arranged to reflect a second light component other than the light component;
A first camera for sensing the first light component to generate a first image; And
A second camera for sensing the second light component to generate a second image;
/ RTI >
According to an exemplary embodiment, the system may further comprise a processing device coupled to the first camera and the second camera, respectively, to generate an overlay image that matches the first image and the second image.
According to an exemplary embodiment, the system may further comprise a filter for removing light having a wavelength band of 600 nm or more from the beam emitted from the second light source.
According to an exemplary embodiment, the system may further include a lens member that collects or guides reflected light emitted from an object to be inspected and transmits the reflected light to the beam splitter.
According to an exemplary embodiment, the beam splitter may be a dichroic mirror.
According to an exemplary embodiment, the first light source may be a narrow band laser, lamp or LED. Specifically, the first light source may be an argon laser, a helium / neon laser, a laser diode, a supercontinuum laser, a pulsed Ti: sapphire laser, a xenon lamp, a mercury lamp, Or a combination thereof.
According to an exemplary embodiment, the second light source may be, for example, a visible light source as a broadband light source. Specifically, the second light source may be an incandescent lamp (a white light tungsten bulb), a halogen lamp, a broadband LED, an OLED, a laser-driven light source (LDLS), or a combination thereof.
According to an exemplary embodiment, in order to extract a beam having a wavelength band of at least 600 nm in front of the first light source, a band pass filter (640/40, etc., long pass filter: 600LP, etc., band pass filter can be installed.
According to an exemplary embodiment, in order to receive only a beam having a wavelength band of at least 600 nm in front of the first camera, and to receive light in a visible light region, filter: 640/40, etc., long pass filter: 600LP, multiple band pass filter) can be installed.
According to a second aspect of the present disclosure,
CLAIMS 1. A method for selectively imaging peripheral nerves in a subject,
Wherein the light source comprises a first light source providing a beam of at least 600 nm wavelength band and a second light source providing a beam of visible light band, , Alternately irradiating the first light source and the second light source to an object to be inspected;
Each of the beams alternately irradiated from the first light source and the second light source is transmitted to a beam splitter through the peripheral nerve through a phenomenon of interference of the peripheral nerve with the reflected light emitted from the object to be examined, Arranged to transmit a first light component while reflecting a second light component other than the first light component; And
Sensing a first light component by a first camera to generate a first image, and sensing a second light component by a second camera to generate a second image;
Is provided.
According to an exemplary embodiment, the method may further comprise processing the first image and the second image to match to produce an overlay image.
According to an exemplary embodiment, the method may further include filtering the beam emitted from the second light source before irradiating the inspection object to remove light having a wavelength band of 600 nm or more. For example, the filter may be a filter (600SP, short pass filter) that passes only a wavelength band of less than 600 nm.
In an exemplary embodiment, the method may further include collecting or guiding the reflected light through the lens member prior to transmitting the reflected light emitted from the inspection object to the beam splitter.
The systems and methods for selectively imaging peripheral nerves in an examination subject provided in accordance with embodiments of the present disclosure may be limited due to problems such as safety, diagnosed only for narrow areas, The limitations of the prior art having the same problem can be effectively solved. Particularly, it is possible to obtain not only a sufficient intensity of reflected light through an interference phenomenon caused by a plurality of layers constituting peripheral nerves but also real-time imaging without using an external marking means such as a fluorescent substance, It has significant advantages in the diagnosis of neuropathy (which may be a side effect of diabetic complications and radiation and chemotherapy), nerve stimulation therapy, and peripheral nerve blocking for peripheral nerves. For example, during the surgical procedure, the sphygmomanometer allows surgery while leaving the peripheral nerve intact during surgery. This can prevent vocal changes due to side effects of thyroid cancer surgery, erectile dysfunction and urinary incontinence caused by side effects of prostate cancer surgery. Therefore, it is expected to be widely used in the future.
1A and 1B are views showing the structural characteristics of the nerve fibers constituting the peripheral nerves and the principle of imaging of the nerve fibers through the principle of thin film interference phenomenon in one embodiment of the present disclosure;
2A and 2B are schematic views schematically illustrating a reflective imaging system and its modification for imaging peripheral nerves during surgery according to an exemplary embodiment of the present disclosure;
FIG. 3A is a schematic diagram schematically showing a system equipped with an integrating sphere device for evaluating optical characteristics of an in vivo part used in Example 1; FIG.
Figure 3b is an optical micrograph showing sampled peripheral nerves, peripheral nerves surrounded by connective tissue, blood vessels and muscles;
4A shows measured values of transmittance and reflectance (20 DEG, 40 DEG and 60 DEG) according to the laser wavelength irradiated under the condition of the incident light power of 10 kHz and the wavelength band interval of 5 nm according to Example 1 Graph;
4B shows measured values of transmittance and reflectance (20 DEG, 40 DEG and 60 DEG) according to the laser wavelength irradiated under the condition of the incident power of 40 Hz and the wavelength interval of 10 nm according to Example 1 Graph;
FIGS. 5A to 5D show that when the wavelength of the excitation laser is set to 640 nm, 561 nm, 488 nm, and 408 nm, respectively, according to Example 2, fat, femoral artery, ), A femoral vein, a femoral nerve, and a sciatic nerve; And
6 is an enlarged image showing the results of a reflection image test of the peripheral nerve during irradiation of a laser having a wavelength of 640 nm.
The present invention can be all accomplished by the following description. The following description should be understood to describe preferred embodiments of the present invention, but the present invention is not necessarily limited thereto.
The accompanying drawings may be somewhat exaggerated in comparison with the thickness (or height) of the actual layer or the ratio with respect to the other layer for the sake of understanding, and the meaning thereof can be appropriately understood by the concrete purpose of the related description to be described later . Further, the details of the individual constitution can be appropriately understood by the concrete purpose of the related description to be described later
The phrases "before" and "at the front end" are understood to be used to refer to the relative position concept. Thus, it is understood that there may be other elements or layers in between, as well as where other elements or layers are directly present in the stated layer. Similarly, the expressions "after" and "at the end" and the phrases "between" Also, the phrase "sequentially" can also be understood as a relative time concept.
"Peripheral nerve" refers to a pathway that transmits sensations collected from the surface of the human body, skeletal muscle, and various internal organs to the central nervous system, and transmits the stimulation of the central nervous system to them again. In the peripheral nerve, there are nerves that transmit sensations and nerves that transmit motor signals. These include the brachial plexus nerve, the common peroneal nerve, the femoral nerve, the lateral femoral cutaneous nerve, the median nerve, Radial nerve, Sciatic nerve, Spinal accessory nerve, Tibial nerve, Ulnar nerve, and so on.
Summary of disclosure
1A and 1B are diagrams showing the structural characteristics of nerve fibers constituting peripheral nerves and the principle of imaging of nerve fibers through the principle of thin film interference development in one embodiment of the present disclosure.
In Fig. 1A, the nerve fibers constituting the peripheral nerve are surrounded by a superficial structure, wherein the myelin is composed of a thin membrane structure composed of a plurality of lipid-rich layers.
As shown in FIG. 1B, the light reflected at the surface of the upper layer and the light reflected at the surface of the lower layer of the thin film structure meet and cause an interference phenomenon, specifically, Constructive interference and destructive interference occur, so that the specific reflection wavelength is enhanced while the other reflection wavelength is suppressed.
As a result, when constructive interference occurs, strong reflected light is emitted, so that a signal of sufficient intensity necessary for imaging can be secured. At this time, the thickness and number of the layers constituting the thin film affect the selectively reflected wavelength, and may be further influenced by the degree of irregularity in the thickness of a few seconds, the composition of the lipid, and other local cell parameters. Therefore, constructive interference can occur in various wavelength ranges of nerve fibers (peripheral nerves) composed of a thin film structure of a plurality of layers, and spectral reflection images are obtained at specific wavelengths by measuring the characteristics obtained therefrom, Can lead to complete nerve fiber imaging.
In this embodiment, it is noted that the reflected light of the specific peripheral nerve can be obtained according to the excitation wavelength of the light to be introduced. That is, in the band above a specific excitation wavelength, as the constructive interference due to the thin film structure of the peripheral nerve becomes noticeable, it can be imaged based on the specific reflected light derived therefrom. On the other hand, I can not get it. Thus, it is possible to selectively or specifically obtain images corresponding to the peripheral nerves.
As such, one embodiment is based on the specific reflection characteristics of the peripheral nerve for excitation light, wherein the excitation light source is at least about 600 nm, specifically about 600 to 750 nm, more specifically about 620 to 700 nm wavelength band Can be used.
In addition, when the intensity of the light source is weak, distinction between peripheral nerves and other tissues (muscles, blood vessels, etc.) may not be easily distinguished. Therefore, It may be advantageous to use a light irradiation of at least about 5 mW / cm < 2 > so as to be able to measure a more pronounced signal.
In one embodiment, the subject to be examined may be part of an animal, for example a mammal, more particularly a part of a human body. In addition, the subject to be examined may be a skin region or subcutaneous tissue region (e.g., a subcutaneous tissue region to be imaged during surgery using surgery and / or an endoscope). More specifically, the surgical site may be a neurosurgical site using a surgical microscope used to observe it.
The imaging method using the above principle is distinguished from the OCT using the interference phenomenon in the prior art in the following points.
OCT is basically a technique that utilizes the interference phenomenon of white light, and utilizes the phenomenon of interference between light reflected from a living tissue and light reflected from a reference mirror. Recently developed OCT technology uses a broadband light source having a bandwidth of 100 nm or more, rather than a short-wavelength light source, although the transmittance of the tissue is increased by using the light of the near-infrared region band rather than the completely white light broad-bandwidth light source, very wide spectrum light source). In this case, the coherence length of the um region is obtained by using a low-coherence light source having a low coherence, thereby increasing the z-axis resolution. As described above, the difference between the path length and the refractive index can be known through interference between the light reflected from the living tissue and the light reflected from the reference mirror. In the case of the time domain OCT, which is the most common form, the position and position change information can be obtained by observing the change of the interference fringe while moving the position of the reference mirror. As a result, Can be obtained. In addition, by detecting only the reflected light corresponding to the path, it is possible to prevent degradation of resolution caused by various other scattered light.
On the other hand, in the case of this embodiment, the light reflected from a specific position (for example, the first layer) and the light reflected from another specific position (for example, the second layer) of the plurality of layers constituting the peripheral nerve Utilize interference and destructive interference. In this case, the difference in the optical path depends on the degree of the light moving in the layer. Particularly, at the surgical site, the surface is usually composed of a liquid (a component containing a large amount of water such as blood and body fluids). In the case of myelin, which is a surface layer of nerve tissue, the surface is composed of lipids, The axial component (axon) is located in the liquid, so that the refractive index of a few seconds is much larger. Therefore, when reflected light appears strongly in the surface layer of the nerve tissue and in the layer in contact with the axon and the axon, and mλ = 2nd cos θ (m = integer, λ = wavelength of the incident light , n = refractive index (a few seconds layer), d = thickness of a few seconds, and [theta] = incident angle of light incident from a few seconds to axon).
In this way, it is possible to acquire images specific to the nerve (especially, peripheral nerve) by selecting a wavelength band in which the degree of reflection is not strong in a main tissue such as blood vessels and muscles in a wavelength causing strong constructive interference.
2A and 2B are schematic diagrams schematically illustrating a reflective imaging system for imaging peripheral nerves during surgery and variations thereof, according to an exemplary embodiment of the present disclosure.
The
And may include two types of light sources, i.e., a
The
The second
Illustratively, the wavelength band of the second light source may range, for example, from about 200 nm to less than 600 nm, specifically from about 350 to 550 nm, more specifically from about 400 to 500 nm.
Optionally, at least one of the
According to the illustrated embodiment, the
Light emitted alternately (or emitted) from each of the
In an exemplary embodiment, for the purposes of beam expansion and uniform illumination, optical fibers, specifically multimode optical fibers, are used to form the first light source or
In the illustrated example, the
In the illustrated embodiment, in the light (reflected light) reflected from the inspection object (for example, the surgical site) alternately illuminated by the first and second light sources, the excitation light from the
The light (reflected light) reflected from the object to be inspected by the alternate irradiation of the
In certain embodiments, when using a zoom lens as the
The reflected light thus collected is filtered through a
According to an exemplary embodiment, light of a wavelength band of approximately 600 nm or more is transmitted (passed) by the
According to an exemplary embodiment, in order to receive only a beam of a wavelength band of at least 600 nm in front of the reflection image camera (first camera), and to receive light of a visible light region in front of the video camera (second camera) A multiple band pass filter such as a long pass filter (640/40 or the like) may be optionally installed (not shown).
On the other hand, it can be understood that the first light component transmitted through the
In the exemplary embodiment, the light (first light component) transmitted by the
In an exemplary embodiment, all of the optical path lengths from the
However, as shown in FIG. 2B, at least one of the
In a particular embodiment, the
In an exemplary embodiment, the first image and the second image generated by each of the
This matched overlay image can be displayed on a display device (not shown). The above-described processing apparatus may include any software and / or hardware adapted to receive an image from each of the
According to a further embodiment, the
In the illustrated image system, the concrete principle of obtaining the overlay image by combining (or matching) the two obtained images can be described as follows.
(Or a first image) and a second light component (or a second image; for example, a visible light image) in the light (reflected light) reflected from the inspection object by the alternate irradiation of the first light source and the second light source, May be obtained alternately at a frequency of, for example, about 10 to 30 Hz, specifically about 15 to 25 Hz, and the two obtained images may be combined by software (for example, a program such as MathWorks's Matlab) Or matched) to obtain an overlay image. For example, visible light is irradiated from the second light source for about 10 to 40 ms (specifically about 20 to 33 ms) to obtain a color image (second image), and then, for example, about 10 to 40 ms (First image) by irradiating excitation light of a specific wavelength band (at least 600 nm) from the first light source for a period of time (specifically about 20 to 33 ms).
Then, the CCD noise image at a predetermined exposure time measured in advance in the reflected image is subtracted, and again, a certain value (approximately 5%) of the maximum intensity value is set as a threshold value, You can remove the value. Then, a process of changing to a pseudo-color image is performed so that it can be easily detected by the naked eye through processes such as median filtering. Through this processing, the neural (peripheral nerve) image can be overlaid on the measured color image to form a new image. Accordingly, a display (e.g., software such as LabVIEW is used) mainly shows three video channels. That is, a color anatomical image, a reflex neural image, and a combined (matched) image of an anatomical image and a neural image can be displayed.
Since the alternation imaging process described above is performed in a sequential manner but is performed at a high speed, it is possible to provide a substantially simultaneous sensed effect in the naked eye, and the matching image may not be subjected to a complicated processing process. Therefore, can do.
Moreover, in the illustrated embodiment, the reflected light has a significantly higher light intensity than the fluorescence image using the conventional fluorescence characteristic, so that the exposure time can be reduced, and furthermore, a highly sensitive detector such as EMCCD can be used So that an image can be obtained quickly.
Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the present invention is not limited thereto.
Example 1
Evaluation of Optical Properties for Biological Tissue Samples
The individual apparatuses constituting the optical property evaluation system of the biological tissue sample used in this embodiment are as follows.
- Laser light source: EQ-99 (Broadband Laser-Driven Light Source from Energetiq Technology, Inc., Spectral Output: 100-1000 nm)
- Spectrometer: Monora 200 of Dongwoo optronics
- Optical chopper: Edmound Optics' Mini Optical Chopper
- current preamplifier: STR 570 from Stanford Research Systems
Lock-in amplifier: a STR 830 from Stanford Research Systems
- integrating sphere: IS236A-4 from Thorlabs
- Photodetector: Thorlabs FDS010
- Control software: Computer LabVIEW (National Instruments, Austin, Texas)
In this example, the optical characteristics of the peripheral nerve in vivo sampled while varying the wavelength of the excitation light source were evaluated, and the optical characteristics of peripheral nerves, blood vessels and muscles surrounded by connective tissues were also evaluated for comparison. FIG. 3A is a schematic diagram of a system equipped with an integrating sphere device for analyzing optical characteristics, and FIG. 3B is an optical microscope photograph of peripheral nerves, peripheral nerves, blood vessels, and muscles surrounded by the sampled connective tissue .
- Principle of Operation of Optical Characterization System
The light emitted from the laser source is selected through a spectrometer composed of a diffraction grating and a slit. The laser light of the selected wavelength is guided through the optical fiber, and the guided laser light is cut at a predetermined timing or frequency by the optical cutter to adjust the timing of the laser light output. The laser beam thus adjusted is introduced through an incident light port into an integrating sphere (a lambertian reflecting surface coated with a material having a high reflectance (white) inside it) so as to capture weak reflected light). At this time, a living body sample is placed at a plurality of points (two in the evaluation system shown) of the outer wall of the integrating sphere so that the incident light transmits and reflects the sample. Specifically, the sample and the reflectance for measuring the transmittance ) Is placed.
The photodetector is a photodiode-based detector that detects reflected light in the integrating sphere with high sensitivity up to 200 to 1100 nm, detects the optical power thereof, (The sensed current is proportional to the light amount of the incident light).
Then, the current (signal) converted by the photodetector is amplified through the signal amplifier, and at the same time, the noise is blocked to prevent the amplification to the noise in the subsequent amplification process in advance. The current signal amplified primarily through the signal amplifier is further amplified by the lock-in amplifier, acquiring and / or storing the spectral data or computer signal by Computer LabVIEW. At this time, Computer LabVIEW also controls the setup of spectroscopy.
- Measurement of transmittance and reflectivity of biotissue samples
Sciatic nerves located in the thighs of rats (Rat, 12 weeks old, male) were sampled, and two samples were prepared, which were enclosed in connective tissue and cleanly removed connective tissue. In addition, blood vessels (vena cava) were collected through surrounding muscle tissue of the thighs and ligature. After the collected sample was placed on a slide glass, water (saline) was slightly dropped to prevent drying, and then the transmittance and the reflectance were measured.
In this embodiment, the above-described optical system for evaluating the optical properties of the integrated spheres (Spectroscopy) was used. The reflected light was measured while varying the incident angle at 20 °, 40 ° and 60 ° over the 400-800 nm or 300-1000 nm wavelength band Respectively. In addition, the intensity of incident light (10 kHz and 40 Hz) and the interval between wavelengths (5 nm and 10 nm) were also measured.
(a) transmittance and reflectance of peripheral nerves surrounded by peripheral nerves and vascular connective tissues according to laser wavelength changes irradiated under the condition of incident power of 10 kHz and an interval of 5 nm, , 40 ° and 60 °), and (b) the intensity of incident light at 40 Hz and the wavelength interval of 10 nm. Measured values of transmittance and reflectance (20 DEG, 40 DEG and 60 DEG) are shown in Figs. 4A and 4B, respectively. In addition, the permeability and reflectance measured for each of the blood vessels and muscles as a control group are also shown in Figs. 4A and 4B.
As shown in the figure, the wavelength band that exhibits highly reflected light in the nerve tissue was about 600 to 700 nm (i.e., 600 nm or more) and showed a remarkably high reflection characteristic in the wavelength band as compared with the muscle tissue and blood vessels in the control group . Also, the peak wavelength was shown in a wavelength band of about 470 to 480 nm, and the actual reflected light intensity at about 550 to 580 nm was not higher than other wavelength ranges, but was somewhat higher than that of muscle tissue and blood vessels.
The reason why the reflected light intensity can be obtained at a wavelength band of at least 600 nm is considered to be that the reflected light of high intensity is emitted from the peripheral nerve by the constructive interference phenomenon in the plural layers constituting the peripheral nerve .
Example 2
Reflection image test according to wavelength of excitation light source
In this embodiment, images are obtained at a plurality of candidate wavelength ranges in order to analyze the reflection image at a specific wavelength band and to select the wavelength band of the excitation light suitable for imaging of the peripheral nerve. At this time, 640 nm, 561 nm, 488 nm and 408 nm were set as a plurality of candidate wavelength ranges (Ex), respectively.
Specifically, a confocal laser scanning microscope (Nikon, A1 Rsi) was used. The tissue to be analyzed was fat, femoral artery, muscle, femoral vein, femoral nerve (femoral nerve) and sciatic nerve (sciatic nerve) were used. For each living tissue, a specific laser was used to irradiate only a single wavelength. In addition, all the reflected light was collected (Em (emission): No filter) without using a separate filter, and the detector was increased in sensitivity using a photomultiplier tube (PMT).
The sciatic nerve and thigh femoral nerve located in the thigh of rats (male, 12 weeks old) were collected and the blood vessels collected through peripheral muscle tissue, adipose tissue, and ligature Femoral vein and femoral artery) were evaluated as controls.
Individual samples were placed on a slide glass, and water (saline) was slightly dropped to prevent drying, and then covered with a cover glass. The reflection images of the fat tissue, the femoral artery, the muscle, the femoral vein, the femoral nerve and the sciatic nerve obtained from the laser light of the excitation wavelength are shown in Figs. 5A to 5D.
According to FIG. 5A, in the case of the femoral nerve and sciatic nerve corresponding to the peripheral nerve at the excitation wavelength of 640 nm, the reflected light appears to be strongest as compared with other tissues. In particular, the magnified reflection image of the femoral nerve and the sciatic nerve shown in FIG. 6 reveals that particularly strong reflected light is emitted from the axon surrounded by aqueducts in the peripheral nerve.
On the other hand, according to Figs. 5B to 5D, there was no specific reflection light for the fat, femoral artery, muscle, femoral vein, femoral nerve and sciatic nerve under test at the excitation wavelength of less than 600 nm.
In order to derive the excitation wavelength band for emitting the specific reflected light to the peripheral nerve, the wavelength of 600 nm or more (specifically, 600 to 700 nm) which was considered as the most probable candidate excitation wavelength band from the spectroscopic data of Example 1 Strong reflection image was obtained at 640 nm belonging to the band.
It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (21)
A light source arranged to illuminate the object to be inspected, wherein the light source comprises a first light source providing a beam of at least 600 nm wavelength band and a second light source providing a beam of visible light band, Wherein the first light source and the second light source are configured to alternately illuminate an inspection target;
Each of the beams alternately irradiated from the first light source and the second light source transmits a first light component indicative of a peripheral nerve among reflected light emitted from the object to be inspected through a peripheral nerve membrane phenomenon of the peripheral nerve, A beam splitter arranged to reflect a second light component other than the light component;
A first camera for sensing the first light component to generate a first image; And
A second camera for sensing the second light component to generate a second image;
/ RTI >
Wherein the light source comprises a first light source providing a beam of at least 600 nm wavelength band and a second light source providing a beam of visible light band, , Alternately irradiating the first light source and the second light source to an object to be inspected;
Each of the beams alternately irradiated from the first light source and the second light source is transmitted to a beam splitter through the peripheral nerve through a phenomenon of interference of the peripheral nerve with the reflected light emitted from the object to be examined, Arranged to transmit a first light component while reflecting a second light component other than the first light component; And
Sensing a first light component by a first camera to generate a first image, and sensing a second light component by a second camera to generate a second image;
≪ / RTI >
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PCT/KR2017/001823 WO2017142376A1 (en) | 2016-02-18 | 2017-02-20 | Non-label imaging system for selective microscopy of peripheral nerve |
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KR102142861B1 (en) * | 2020-05-26 | 2020-08-10 | 주식회사 에프앤디파트너스 | Dermascope device capable of real-time observation and shooting at the same time |
WO2021241795A1 (en) * | 2020-05-26 | 2021-12-02 | 주식회사 에프앤디파트너스 | Dermatoscope device capable of simultaneous real-time observation and image capturing |
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