Apparatus and method for detecting fluorescence

Abstract

Disclosed is an apparatus (100) for detecting fluorescence of a sample (118, 119) obtained from a subject, the sample (118, 119) containing one or more types of fluorophores and indicative of a disease or condition, the sample (118, 119) being conveyed from the subject in a transparent conduit (131). The apparatus includes a housing (10) with one or more light sources (110) and one or more light receivers (120), and a computing device (180). Also disclosed is a method for detecting fluorescence of a sample (118, 119) obtained from a subject, the sample (118, 119) containing one or more types of fluorophores and indicative of a disease or condition, the sample (118, 119) being conveyed from the subject in a transparent conduit (131). Additionally disclosed are uses of the apparatus (100) and a kit of parts including the apparatus (100).

Description

Technical Field

[0001] The present disclosure (the present invention) generally relates to the detection of fluorescence of a sample. In particular, the present invention relates to an apparatus for detecting the fluorescence of a sample containing one or more fluorophores (also referred to as fluorescent generating substances, fluorescent dye molecules, or phosphors), and a method for detecting the fluorescence of a sample containing one or more fluorophores.

Background Art

[0002] Fluorescence-guided photodynamic substances, such as 5-aminolevulinic acid (5-ALA), are increasingly used in surgical procedures or operations for tumors, such as gliomas of grade (malignancy) 3 to 4. Fluorescent markers and fluorophores emit fluorescence when excited at a specific wavelength, and emit characteristic fluorescence that helps surgeons, for example, when identifying the infiltration zone between tumor cells and healthy tissue. In current methods, detection is based on fluorescence visible to the surgeon, and the average tumor removal rate is 80%. It has been found that there is an optimal benefit for patients with at least 98% of the tumor removed and no damage to functionally important brain tissue. Tumors are mechanically removed mainly by using various surgical methods, such as removal of tissue by an ultrasonic aspirator or a surgical suction instrument after manual dissection of the tumor. Using currently available surgical removal techniques, the resection rate is not sufficient, and the evaluation of the boundary of healthy tissue is unreliable.

[0003] Neurosurgeons use, for example, the agent 5-ALA, which is 5-aminolevulinic acid or a salt thereof, such as the hydrochloride salt, in fluorescence-guided surgery (FGS) of certain types of glioma brain tumors. Prior to surgery, the patient is orally administered the 5-ALA agent. 5-ALA is a compound in the porphyrin synthesis pathway, which in the human body is converted to protoporphyrin IX (PpIX), a fluorescent substance (fluorophore), and further to heme by ferrochelatase. Thus, 5-ALA may be considered a prodrug of PpIX. Cancer cells have little or reduced ferrochelatase activity, and as a result, among other potential reasons, there is an accumulation of PpIX in cancer cells. During FGS, the surgical cavity is exposed to PpIX, which exhibits blue light, causing the cancer cells of the tumor to fluoresce at a peak wavelength of approximately 635 nm (occurring with PpIX), and this fluorescence helps to delineate the tumor for its removal. The fluorescence of PpIX (and the 635 nm peak wavelength) is not visible under white light, but under blue excitation light, the tumor containing PpIX stands out compared to others. Thus, in FGS, to see the fluorescence, the surgeon changes the light emitted by the microscope from white to blue. However, when the surgical cavity is irradiated with the blue light that excites PpIX, the contrast with other tissues (excluding the tumor) is significantly reduced. Thus, while the cavity emits blue light, it is difficult for the surgeon to distinguish living or important tissues that should not be removed. Therefore, tumor removal is mainly performed using white light, but its identification is carried out using blue excitation light.

[0004] Thus, fluorescence detection is restricted by the vision of the surgeon. Modern FGS technology mainly depends on the subjective evaluation of perceptible fluorescent traces, which is considered likely to cause both type 1 and type 2 errors. Parallel to the required manual configuration of the light source, modern FGS delays the performance during the operation. Furthermore, during tumor removal, the visual function is restricted, especially by the limited field of view of the surgical microscope, invisible residues, and angles. Due to technical constraints, the removal method is determined by the memory of the observed fluorescence deposits and, in some cases, the optically filtered video recording. Also, the human visual system lacks the sensitivity to detect clinically relevant concentrations of fluorophores, such as PpIX, and there are limits to visual specificity. By exposing a sample containing a fluorophore to excitation light, the fluorescence fades over time due to photochromism, which is determined by both the excitation time and the intensity of the excitation light. Photobleaching occurs, for example, due to the reaction between the fluorophore and surrounding molecules, thus restricting the use of fluorophores.

[0005] Several other experimental optical imaging techniques, such as Raman spectroscopy, optical coherence tomography (OCT), and diffuse reflectance spectroscopy (DRS), have been developed for neurosurgical applications. Most of the above-mentioned optical imaging methods are expected to be suitable for the boundary delineation and functional measurement of neurosurgical tumors in current scientific and technical literature. However, their current intraoperative applications mainly require separate bulky sensors and complex visualization methods, which impair the skills of the surgeon.

Summary of the Invention

Problems to be Solved by the Invention

[0006] Therefore, in light of the above description, there is a need to solve the above-mentioned drawbacks related to the detection of fluorescence of a sample containing one or more fluorophores and representing a disease or pathological condition.

Means for Solving the Problems

[0007] According to one aspect, a subject matter of an independent claim is provided. Embodiments are described in dependent claims. The protection scope of various embodiments of the present invention is defined by the independent claim. Embodiments, examples, and features (if any) described herein that do not fall within the scope of the present invention described in the independent claim should be construed as useful examples for understanding various embodiments of the present invention.

[0008] The present invention aims to provide an apparatus for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores, representing a disease or pathological condition, and being delivered from the subject through a transparent conduit. The present invention also aims to provide a method for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores, representing a disease or pathological condition, and being delivered from the subject through a transparent conduit. The present invention aims to provide a solution to existing problems in detecting fluorescence of a sample containing one or more fluorophores, especially during fluorescence-guided surgery. The object of the present invention is to provide a solution that at least partially solves the problems found in the prior art and improves the technology.

[0009] In one aspect, an embodiment of the present invention is an apparatus for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores, representing a disease or pathological condition, and being delivered from the subject through a transparent conduit, the apparatus comprising - a housing configured to surround at least a part of the transparent conduit, the housing comprising - one or more light sources operable to emit light towards the sample carried through the transparent conduit, - one or more light receivers operable to detect fluorescence generated by one or more fluorophores in the sample carried through the transparent conduit, and - a computing device, the computing device comprising -Receiving a first signal from at least one of the one or more light receivers, the first signal being proportional to the fluorescence detected by at least one of the one or more light receivers, -Comparing the first signal with a predetermined threshold value for the first signal, and -Providing an apparatus characterized by having computing means for outputting first information representing the comparison result.

[0010] In another aspect, one embodiment of the present invention is a method for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores and representing a disease or pathological condition, the sample being transported from the subject by a transparent conduit, the method comprising: i) transporting a sample containing one or more fluorophores and representing a disease or pathological condition from the subject by a transparent conduit; ii) emitting light towards the sample being transported through the conduit; iii) detecting the fluorescence generated by one or more fluorophores in the sample being transported through the conduit; iv) generating a first signal proportional to the detected fluorescence; v) comparing the first signal with a predetermined threshold value for the first signal; vi) outputting first information representing the comparison result. A method is provided that includes the steps.

[0011] In another aspect, one embodiment of the present invention provides the use of the apparatus disclosed herein for performing a method selected from the group consisting of invasive medical methods, methods for detecting diseased tissue or diseased body fluids containing one or more fluorophores and representing a disease or pathological condition, methods for detecting tumors, and methods for diagnosing cancer. Preferably, the diagnosis of cancer is an in vitro or in vitro diagnosis of cancer.

[0012] In another aspect, one embodiment of the present invention provides a component kit that combines the apparatus disclosed herein with a transparent conduit.

[0013] In another aspect, one embodiment of the present invention provides a surgical method, a method for treating a disease or a medical condition, or an in vivo diagnosis of a disease or a medical condition, using the apparatus disclosed herein.

[0014] In another aspect, one embodiment of the present invention provides a method for detecting fluorescence of a sample containing one or more fluorophores and representing a disease or a medical condition, using the apparatus disclosed herein.

[0015] In another aspect, one embodiment of the present invention provides the apparatus disclosed herein for carrying out the method disclosed herein.

[0016] Embodiments of the present invention substantially eliminate or at least partially solve the above problems in the prior art of conventional fluorescence detection devices and methods, and embodiments of the present invention enable the detection of low-concentration traces that are not visible by fluorescence (invisible to the human eye).

[0017] Additional aspects, additional advantages, additional features, and additional objects of the present invention will become apparent from the following drawings and detailed description of exemplary embodiments, which are to be construed in connection with the claims appended hereto.

[0018] As will be appreciated, the features of the present invention are combinable with each other in various combinations without departing from the scope of the present invention as set forth in the claims.

[0019] The above summary of the invention and the following detailed description of exemplary embodiments are well understood when read in conjunction with the accompanying drawings. For the purpose of explaining the invention, an exemplary configuration of the present invention is shown in the drawings. However, the present invention is not limited to the specific methods and apparatuses disclosed herein. Further, as will be understood by those skilled in the art, the drawings are not drawn to scale unless otherwise specified. Whenever possible, the same elements are denoted by the same reference numerals.

[0020] Next, embodiments of the present invention will be described below with reference to the accompanying drawings, which are merely illustrative.

Brief Description of the Drawings

[0021]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Modes for Carrying Out the Invention

[0022] In the accompanying drawings, underlined reference numerals are used to represent the item indicated by the underlined reference numeral or an item located adjacent to the underlined reference numeral. Non-underlined reference numerals relate to items specified by lines connecting the non-underlined reference numerals to the items. When a reference numeral is not underlined and is accompanied by a relevant arrow, the non-underlined reference numeral is used to identify a general item indicated by the arrow.

[0023] The following detailed description shows embodiments of the present invention and ways to implement these embodiments. Although several embodiments of practicing the present invention are disclosed, as will be recognized by those skilled in the art, other embodiments of practicing or embodying the present invention are also possible.

[0024] In one aspect, an embodiment of the present invention is a device for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores and representing a disease or pathological condition, the sample being delivered from the subject through a transparent conduit, and the device comprising: - a housing configured to surround at least a portion of the transparent conduit, the housing comprising: - one or more light sources operable to emit light towards the sample carried through the transparent conduit; - one or more light receivers operable to detect fluorescence generated by one or more fluorophores in the sample carried through the transparent conduit; - a computing device, the computing device comprising: - receiving a first signal from at least one of the one or more light receivers, the first signal being proportional to the fluorescence detected by at least one of the one or more light receivers; - comparing the first signal with a predetermined threshold value for the first signal, and - providing a device characterized by having computing means for outputting first information representing the result of the comparison.

[0025] Accordingly, the device of the present invention enables the detection of fluorescence of a sample obtained from and carried out of a subject, and in particular enables the detection of the sample almost in real time. The expression "almost real-time detection" used herein and hereinafter means the time between the time when a sample is obtained from a subject and the time when first information representing a comparison result obtained by comparing a first signal (this signal is proportional to the fluorescence detected by at least one of one or more light receivers) with a predetermined threshold value for this signal is output. Almost real time is less than 10 s, preferably less than 5 s, more preferably less than 2 s, even more preferably less than 1 s, more preferably less than 0.1 s, and most preferably less than 0.05 s. One advantage of the device of the present invention for detecting the fluorescence of a sample is that it can provide the user with real-time feedback on the fluorescence of the fluorophore in the sample obtained from the subject. Another advantage of the device of the present invention is that it can detect fluorophores that cannot be detected by the eye if not configured as described above, that is, fluorophore concentrations that are too low for visual detection (clinically significant amounts of fluorophores that are too low for visual detection) or wavelengths of fluorescence of fluorophores that cannot be detected by the human eye. In particular, the device of the present invention can provide guidance to a surgeon during FGS, whereby tissues that are not so life-threatening or important to the subject (human or animal) can be removed, because after emitting light (e.g., blue light when exciting PpIX) that excites the fluorophore into the surgical cavity of the subject and applying it, in order to visually detect the portion to be removed in the surgical cavity due to the generated fluorescence, the surgeon can switch the wavelength of the light source to white light and remove a sample containing one or more fluorophores and representing a disease or pathological condition. When the surgeon removes a sample (e.g., tissue not representing a disease or pathological condition) that generates fluorescence below a predetermined threshold value of fluorescence, the surgeon is provided with feedback from the device based on the output information.Next, the surgeon can stop removing any further samples from the surgical cavity, thus preserving tissue that is critical to the patient's life. Therefore, the device of the present invention can identify non-fluorescent regions of the surgical cavity and minimize damage to healthy tissue. Additionally, the device of the present invention enables near real-time detection of fluorescence during surgery and can speed up the surgeon's intraoperative actions.

[0026] Furthermore, the device of the present invention emits light that excites one or more fluorophores of the sample being carried out from the subject through a transparent conduit, so that the excitation light emitted towards the surgical cavity can be reduced, thus reducing photobleaching of the fluorophore and subsequently enabling a longer surgical time.

[0027] The advantages and technical effects when using fluorescence-based detection according to the present invention can be summarized as an increase in sensitivity, accuracy, and detection speed compared to other detection methods. The increase in sensitivity means that fluorescence-based detection is highly sensitive and can detect even a tiny amount of fluorescence in the sample. The increase in accuracy also means that in this case, the detection is extremely specific in detecting only the sample that induces the characteristic fluorescence. Fluorescence in the form of fluorophores is induced in the sample, for example, cancer tissue, before detection. Therefore, only the tissue or sample that is fluorescent and thus detectable is detected, and thus the detection is extremely specific to the sample or tissue of interest. The detection speed is also important in this case because it enables a rapid response due to the detected changes in fluorescence.

[0028] Specifically, the accuracy and speed of detection of the present invention enable the analysis of not only aerosols or vapors but also flowing tissues, i.e., laminar and turbulent tissues. Due to the sensitivity of fluorescence, the targeted fluorophore can be detected deep inside the solution. The detection is not affected by the composition of the sample to be analyzed (detected), and the device is also suitable for analyzing samples such as, for example, mixtures of liquids, tissues, and aerosols. In addition, all samples, including macroscopic and microscopic sizes carried in transparent conduits, are analyzed and detected for fluorescence, not just for some specific samples, such as aerosols.

[0029] It is appropriate to understand that fluorescence is not the spectral reflection of light and that light is not only a wave phenomenon but also a particle. By using fluorescence instead of spectral reflection or spectral absorption and photons instead of wave patterns, the high sensitivity and high flexibility of the present invention can be obtained when analyzing tissues in the conduit. The high sensitivity is brought about by the above-mentioned insight that fluorescence is essentially a light source in the conduit and can be detected from a direction without any interference, rather than being reflected or absorbed. Furthermore, fluorescence produces secondary effects, such as heat and vibration, which can be detected, for example, by a wave detector. By combining the information from the photon detector and the wave detector, an increase in sensitivity can be obtained for specific conditions, and it can be said that one does not exclude the other. In this technical concept, for example, in wave optoacoustic imaging or Raman spectroscopy, by using a sensor that is not sensitive to photons at a specific wavelength, it is appropriate to understand that the analysis can be complemented by detecting secondary effects called "fluorescence interference", particularly heat generation and vibration. Also, fluorescence is characterized by its half-life called "fluorescence decay", and this fluorescence decay can be altered by the characteristics of the mixture, such as temperature, pH, and minor tissue components, in addition to the fluorescence peak wavelength.

[0030] As an additional advantage, altering the components of the mixture according to the present invention can increase the sensitivity of the detector, and such alteration can be carried out, for example, by adjusting the tissue perfusion fluid. These slight features related to fluorescence result in much higher flexibility with respect to medical applications than applications using non-fluorescence.

[0031] In the present invention, the difference between fluorescence lifetimes can be detected not by a stand-alone probe placed on the tissue, but due to the laminar flow model in the conduit and the flow of the sample. In contrast to a probe from one angle, analyzing the fluorescence characteristics of the sample in the conduit enables the achievement of very high sensitivity, because it can be achieved up to a level that may be harmful when applied to living tissue for the amount of photoexcitation and emission. Since the placement of both the light source and the detector is free at 360°, and can be placed at any angle around the conduit, the obstruction of the excitation light to the target and the emission light from the target can be minimized. In addition, due to the black box design that can be implemented by the post-excision design, a good noise-to-signal ratio can be obtained in contrast to the probe used in non-excised living tissue contaminated by other light sources.

[0032] The present invention is based on the detection of fluorescence, i.e., fluorescence emission, fluorescence interference, or fluorescence decay, derived from a target tissue induced with a specific fluorophore. Thereby, extremely specific and accurate tissue-specific detection is possible. In addition, there is no need to "teach the system" because fluorescence is based on a predetermined fluorophore with known fluorescence characteristics. A database of signals and outputs for various tissues is unnecessary. Thereby, the detection depends not on the tissue itself but on a predetermined fluorophore induced for the target tissue. Since the detection is fluorophore-specific detection rather than tissue-specific detection, interference from the environment or other tissues other than the target tissue does not occur. The detection is not interfered with by contaminants, disturbances, or other tissues because these are not specified as monitored characteristics. However, even when the detection does not necessarily require "learning" or comparison with a database, since the fluorophore is predetermined, learning based on the database can further improve performance.

[0033] The device of the present invention can be integrated into existing surgical instruments, such as ultrasonic aspirators or surgical suction instruments, without interfering with their conventional use. In addition, the device can be integrated into a surgical system that monitors the inflow and outflow of a fluorescence probe. Such information can be used to optimize the timing, effectiveness, or detection of adverse events associated with a fluorescent target molecule that may represent a disease or health condition.

[0034] The device does not necessarily require additional extra equipment for detection, such as a suction tip, specific forceps, or in vivo probe, substance, or solution. Instead, the device itself is a stand-alone device that complements the surgical workflow.

[0035] The housing of the device of the present invention is adapted to surround at least a part of the transparent conduit through which the sample is carried away from the subject, so that the drawbacks in the case of using a conventional separate bulky or hand-held device for detecting fluorescence are solved especially during FGS, because the present device can be attached to the conduit. The device of the present invention can be arranged extremely close to the subject, for example, at a distance of 1 to 100 cm from the subject, or at a further distance from the subject, for example, at a distance of 1 to 10 m from the subject, as seen from the location where the sample is obtained. The distance between the housing of the device and the subject is not limited as long as the sample is being carried through the transparent conduit. When near real-time detection is required, the distance from the housing to the subject is preferably short enough to enable near real-time feedback to the user of the present device, and the housing does not significantly and negatively interfere with the user's work. Preferably, the housing surrounds the transparent conduit at a distance of 0.01 to 10 m, preferably 0.3 to 10 m, more preferably 0.3 to 3 m, and even more preferably 0.3 to 1 m from the subject from which the sample is obtained. Therefore, the device of the present invention is not restricted to only a short distance from the subject.

[0036] In particular, the device of the present invention improves the ability of the surgeon to identify cancerous tissue in a way that reduces the current technical constraints affecting FGS, by, for example, comparing the detected fluorescence with a threshold or a determined value, not only by the human eye but also by a detection method based on the use of spectral analysis. According to the results shown by the entire study, computer-based mechanical classification of healthy and diseased tissues is more sensitive and better than the subjective performance of the human eye. By using the method of the present invention, optical spectrum analysis can also recognize wavelengths of light that cannot be detected by the eye (for example, ultraviolet, near infrared, infrared), beyond the physiological limits of the human eye.

[0037] Thus, the device of the present invention for detecting the fluorescence of a sample being carried within, for example, a surgical suction instrument has the advantages of improving patient care, delaying and reducing tumor recurrence, and enhancing patient safety. With the device of the present invention, it is possible to detect the fluorescence of a sample containing one or more fluorophores and representing a disease or condition (such as a tumor) that is not visually detectable to the human eye. Thus, postoperative complications are reduced, and the cost of healthcare management is reduced by the reduction of postoperative complications and reoperations adjacent to damage to eloquent brain areas.

[0038] As used herein and hereinafter, the term "fluorescence" means the emission of light by atoms, molecules, nanostructures, fluorophores, substances, or samples that have absorbed light or other electromagnetic radiation. It should be understood that fluorescence is a form of luminescence. In fluorescence, often the emitted light has a longer wavelength than the absorbed radiation and thus has lower energy. Fluorescence occurs when a molecule, atom, nanostructure, fluorophore, substance, or sample absorbs light or other electromagnetic radiation and is excited and then transitions to a lower energy state by the emission of one or more photons. Fluorescent atoms, molecules, nanostructures, substances, and samples are fluorophores. It should be understood that different fluorescent molecules (fluorophores) and atoms can be excited by light of different wavelengths and that fluorescent molecules (fluorophores) and atoms can emit light of different wavelengths.

[0039] As used herein and hereinafter, the term "sample" means a part or piece obtained from a substance. Examples of samples include, but are not limited to, tissues such as connective tissue, muscle tissue, nerve tissue, and epithelial tissue, tumors such as brain tumors such as glioma-type tumors and diffuse cancer types such as carcinoma and sarcoma, intracellular and extracellular fluids such as cerebrospinal fluid, urine, and blood, blood cells, and extracellular vesicles.

[0040] As used herein and hereinafter, the expression "indicating a disease or pathological condition" means that a sample obtained from a subject has a disease or pathological condition, and in this case, a sample containing one or more fluorophores is an indicator of the disease or pathological condition. Thus, it should be understood that if such a sample contains one or more fluorophores, this sample may indicate a disease or pathological condition. Examples of diseases and pathological conditions include cancer, such as glioma, non-functional pituitary adenoma, carcinoma, sarcoma, benign neoplasm, non-invasive neoplasm, malignant neoplasm, bacterial infection caused by, for example, Escherichia coli, viral infection, deoxygenation, cerebrospinal fluid leakage, contamination of body fluids by harmful or toxic compounds, and drug accumulation or systemic clearance, but are not limited thereto. It should be understood that fluorescence may be caused by at least one of one or more fluorophores derived from fluorophores in the sample or autofluorescence of fluorophores (e.g., NAP(P)H, FAD, flavin, collagen, vitamins, such as vitamin A1, vitamin B2, vitamin B6, and B9, indoleamine), mitochondria, or drugs that can function as lysosomes (e.g., indocyanine green, gliolan, i.e., 5-ALA hydrochloride), prodrugs (e.g., methyl ester of 5-ALA, dipeptide derivative of 5-ALA), precursors or any other suitable compounds, and fluorescence may occur due to one or more fluorophores in a sample indicating a disease or pathological condition.

[0041] As used herein and hereinafter, the term "subject" means a human or an animal. Thus, "a sample obtained from a subject" means a sample obtained from a human or an animal, for example, by surgery or an invasive medical procedure.

[0042] As used herein and hereinafter, the term "fluorophore" means a fluorescent molecule, atom, nanostructure, fluorophore, or substance that can re-emit light upon photoexcitation, i.e., during fluorescence. The fluorophore may or may not be covalently bound to the sample by one or more chemical bonds. Examples of fluorophores include PpIX and its salts and derivatives, indocyanine green and its salts, methylene blue, fluorescein and its salts (e.g., sodium fluorescein, etc.), cyanines (e.g., Cy5.5, Cy7, Cy7.5), T700, T800, BLZ-100, GB119, IRDye800CW conjugate, IRDye700DX conjugate, EC17, LUM015, AVB-620, folic acid-fluorescein isothiocyanate (FITC), OTL38, gGlu-HMRG, green fluorophore conjugate, fluorescent dye-labeled peptide, fluorophore-conjugated antibody, fluorescent nanoparticles, activatable fluorescent probes, endogenous fluorophores, NAP(P)H, FAD, flavin, collagen, vitamins such as vitamin A1, vitamin B2, vitamin B6, and vitamin B9, indoleamine, mitochondria and lysosomes, and their derivatives, isomers and salts, or combinations thereof, but are not limited thereto. It should be understood that one or more fluorophores in the sample may function as fluorophores that may or may not be converted to fluorophores during biosynthesis or by reaction and may be derived from drugs, prodrugs, precursors, or any suitable compound. Thus, the above-mentioned drugs, prodrugs, precursors, or any other suitable compound that can function as a fluorophore may be converted to various derivatives of the fluorophore. Examples of precursors / drugs that are converted to fluorophores include the photosensitizing agent 5-ALA and its salts, preferably its hydrochloride salt, which are converted to fluorescent PpIX and its derivatives in the human body. A fluorescent marker or tag may also be a fluorophore.

[0043] As used herein and hereinafter used in combination with "sample obtained from a subject and delivered from the subject through a transparent conduit", the expression "sample obtained from a subject" means a sample obtained from a human or animal by, for example, surgery or an invasive medical treatment method, and this expression preferably means that the sample is obtained and at the same time, instantaneously or immediately after being delivered from the subject through a transparent conduit. As used herein and hereinafter used, the expression "delivered" means, for example, the movement of a sample moving away from a subject by a suction device, for example, by negative pressure, and examples of such a suction device include, but are not limited to, an ultrasonic aspirator or a surgical aspirator. Gravity can also be used to deliver the sample from the subject. Therefore, it should be understood that the device for detecting the fluorescence of a sample disclosed herein and hereinafter disclosed can detect the fluorescence of a sample in a state where the sample is delivered from a subject, that is, a moving sample, through a transparent conduit. As used herein and hereinafter used, the expression "transparent conduit" means a conduit through which light can pass through the material of the conduit. Examples of conduits include, but are not limited to, pipes, tubes, such as suction tubes, surgical suction tubes, medical tubes, hygienically sensitive tubes, catheters, cannulas, and suction tips. The outer diameter of the conduit is not limited as long as the conduit can be used in surgery to deliver a sample from a subject. For example, the outer diameter of the conduit can be 1 mm (3 Fr) to 60 mm (180 Fr), preferably 1 mm (3 Fr) to 20 mm (60 Fr), more preferably 1 mm (3 Fr) to 16 mm (48 Fr), even more preferably 2 mm (6 Fr) to 16 mm (48 Fr), and even more preferably the outer diameter of a surgical suction tube or a medical tube of 2.67 mm (8 Fr) to 16 mm (48 Fr). The inner diameter of the conduit having such an outer diameter can be an inner diameter that can carry a sample inside the conduit and can be used for a medical treatment. For example, the inner diameter of the conduit is preferably 0.75 mm to 15.75 mm.Examples of samples obtained from a subject and carried out of the subject via a transparent conduit are portions of a tumor removed from a human with an ultrasonic cavitation device (ultrasonic aspirator), carried into a medical tube from the human simultaneously with the removal, and further removed from the human.

[0044] As used herein and hereinafter, the term "housing" means a structure that includes one or more light sources and one or more light receivers (detectors), and these light sources and light receivers are arranged in horizontal, vertical or angled positions within this housing. The housing is adapted to surround at least a portion of a transparent conduit. One or more light sources are preferably arranged to emit light towards a sample being carried within the transparent conduit and are operable. One or more light receivers are preferably arranged to detect fluorescence generated by one or more fluorophores in a sample being carried within the transparent conduit and are capable of operating. Optionally, in addition to one or more light sources and one or more light receivers, the housing has at least a partially enclosed compartment having a top section, a bottom section of the housing adapted to surround at least a portion of the transparent conduit, and openings provided at opposite longitudinal ends of the housing. The housing can reduce the entry of external light to the light receivers and can prevent the light sources and light receivers from getting dirty. Additionally, the housing adapted to surround the transparent conduit allows the housing to be removably fixed to the conduit, such that the user can fix the housing to the conduit and use their hands for other things, and the user can remove the housing from the conduit when the device is not in use. The housing may further have attachment means for attaching and / or locking one or more components that can form the housing and / or for externally attaching to the conduit, such as nails, clips, vacuum suction cups, screws, hooks, combinations of bolts and nuts, brackets, locks, tapes, snap locks, hinges, seals, one or more magnets, etc. It should be understood that as used herein and hereinafter, the term "surround" means that the housing at least partially surrounds or completely surrounds the conduit, and the expression "at least a portion of the transparent conduit" as used herein and hereinafter means that the housing surrounds a part or the whole of the conduit, i.e., with respect to the longitudinal axis of the conduit.Furthermore, it should be understood that the housing may be configured to be removable so as to surround at least a portion of the transparent conduit. Further, the housing may be removably fixed to the conduit, for example, with seals located at opposite longitudinal ends of the housing, or by the housing clamping the conduit. In one embodiment, the housing has at least one of a top section, a bottom section, an opening, at least one bore, a switch, and others. Further, the switch can be operated to emit light when one or more light sources of the housing are in operation or in the ON mode. Access to the inside of the housing is possible through at least one bore, for example, by a power supply device, operation, data transfer, etc. In one example, the housing is arranged through at least one bore and includes a power supply device for supplying power to the housing. The power supply device can supply power using conventional methods, and such methods include, but are not limited to, solar energy, electrical energy, chemical energy, batteries, rechargeable batteries, fuel-based energy, hydroelectric power, and others. In another example, the housing may be arranged through at least one charger and include a power supply device for supplying power to the housing. The charger can wirelessly draw power from an electromagnetic field generated by a transmitter device driven by power from a power supply source, that is, the power is supplied to the housing by a wireless power transmission system. It should be understood that the computing device may be provided inside the housing or outside the housing.

[0045] As used herein and hereinafter, the term "light source" means a device that emits light. The housing can have one or more, for example, one, two, three, or four, or more light sources. Each light source can emit light of the same or different wavelengths, or a range of wavelengths. Specifically, the light source is operable to emit light toward a sample being carried within a transparent conduit. The light source can emit light within a predetermined wavelength range. Examples of light sources include, but are not limited to, light-emitting diodes (LEDs), laser diodes, halogen lamps, incandescent lamps, fluorescent lamps, and LEDs combined with quantum dots, or combinations thereof. In a preferred embodiment, at least one of the light sources is an LED, although this can be any other suitable light source, i.e., a light source that can emit light within a desired narrow wavelength range and at a required intensity, such as a laser diode. In one embodiment, the housing includes a plurality of light sources, preferably LEDs, for example at least two LEDs, electrically coupled to a power supply device disposed within the housing.

[0046] As used herein and hereinafter, the term "light receiver" means a photodetector that detects light. One or more light receivers are operable to detect fluorescence generated by one or more fluorophores in a sample being carried within a transparent conduit. Specifically, the light receiver is preferably operable to detect light of a predetermined wavelength or wavelength range. Examples of light receivers include, but are not limited to, image sensors, such as charge-coupled device (CCD) sensors (e.g., BT-CCD image sensors, CCD linear image sensors), and metal-oxide-semiconductor (MOS) sensors (e.g., MOS linear image sensors, complementary MOS (CMOS) sensors), photodiodes, such as avalanche photodiodes (APDs), and phototransistors, or any combination thereof.

[0047] As used herein and hereinafter, the term "computing device" means a device that includes computing means, and such computing means includes, but is not limited to, mobile stations (cell phones), smartphones, personal digital assistants (PDAs), handsets, devices using wireless modems (such as alarm or measurement devices), laptops and / or touchscreen computers, tablets, game consoles, notebooks, and multimedia devices. It should be understood that a computing device may be a nearly exclusive uplink-only device, an example of which is a laptop that loads data (corresponding to one or more signals received from at least one of one or more light receivers) onto a network. A computing device may be a device having a function of operating in a mono Internet of Things (IoT) network, which is a scenario having a function that a system can transfer data on a network without requiring interaction from person to person or from person to computer. Additionally or alternatively, a computing device may be a device having computing means for implementing at least some of the methods of explanatory symmetry disclosed herein and hereinafter. Some exemplary computing means for implementing a process may preferably include at least one of the means listed below, and such means includes a data collection unit, a processor (including dual-core processors and multi-core processors), a digital signal processor, a current-voltage converter, an analog-digital converter, an amplifier, an optical interrupter, a controller, a data receiving device, a data transmitting device, an encoder, a comparator, a decoder, a memory, a RAM, a ROM, software, firmware, a display, a user interface, acoustic means such as loudspeakers, LEDs, an augmented reality interface, a user interface, a display circuit, a user interface circuit, user interface software, display software, a circuit, an antenna, an antenna circuit, and an electrical circuit.It should be understood that the computing device may be located externally to the housing or may be provided in the housing, preferably provided within the housing. Further, it should be understood that the computing means may further include output means as means for enabling the output of information to the user, or may be operably connected to this output means. Examples of output means include, but are not limited to, a display, a user interface, acoustic means such as loudspeakers, LEDs, an augmented reality interface, a user interface, a display circuit, a user interface circuit, user interface software, display software, a circuit, an antenna, an antenna circuit, an electrical circuit, etc.

[0048] As used herein and hereinafter, the term "information" means sound, text, light, pressure changes, and data or combinations thereof for the computing means. The information is output by output means operably connected to the computing means.

[0049] In a preferred embodiment, the device is adapted to detect the fluorescence of a sample obtained from a subject substantially in real time, the sample containing one or more fluorophores, representing a disease or medical condition, and the sample being carried away from the subject within a transparent conduit.

[0050] In a preferred embodiment, the one or more light sources and the one or more light receivers are provided within the housing. This makes the device easier to use, as it is possible to avoid the use of external light sources and light receivers and avoid the device becoming complex during use. Additionally, the housing protects the one or more light sources and the one or more light receivers from becoming dirty.

[0051] In one embodiment, the computing device further includes means for measuring the fluorescence level and the output represents the detected fluorescence level.

[0052] In a preferred embodiment, the computing device is connected to, and preferably communicatively coupled to, the housing. In a preferred embodiment, the computing device is communicatively coupled to at least one of the one or more light sources and / or at least one of the one or more light receivers by means of a wired connection means or a wireless connection means. In a particular embodiment, the computing device is communicatively coupled to one or more light receivers provided within the housing.

[0053] In one embodiment, the housing - one or more light sources operable to emit light towards a sample carried in a transparent conduit, - one or more light receivers operable to detect fluorescence generated by one or more fluorophores in a sample carried in a transparent conduit, - the computing device is communicatively coupled to at least one of the one or more light sources and / or at least one of the one or more light receivers, and the computing device - receives a first signal from at least one of the one or more light receivers, the first signal being proportional to the fluorescence detected by at least one of the one or more light receivers, - compares the first signal with a predetermined threshold value for the first signal, and - has computing means for outputting first information representing the result of the comparison.

[0054] Accordingly, it should be understood that the computing device is provided within the housing.

[0055] In one embodiment, the angle formed by the longitudinal central axis of at least one of the one or more light sources and the longitudinal central axis of at least one of the one or more light receivers is selected from 0° to 180°. In one embodiment, this angle is selected from 15° to 180°. In one embodiment, this angle is selected from 90° to 180°, preferably 90°. Different angles can be used to position the one or more light sources and the one or more light receivers at different locations within the housing.

[0056] In one embodiment, the height of the housing is from about 10 mm to about 100 mm, and the width of the housing is from about 10 mm to about 150 mm. In one embodiment, the height of the housing is from about 50 mm to about 70 mm, and the width of the housing is from about 80 mm to about 130 mm. It should be understood that the dimensions of the housing are not limited as long as the housing surrounds at least a portion of the transparent conduit.

[0057] In a preferred embodiment, the computing device includes means for outputting first information regarding the first signal in response to a first signal that exceeds or is equal to a predetermined threshold, and for outputting second information regarding the first signal in response to a first signal that does not exceed or is not equal to the predetermined threshold.

[0058] In one embodiment, the housing is configured to surround the entire periphery of the transparent conduit. This can minimize the external light reaching the one or more light receivers (i.e., the light from the surrounding outside of the housing), and thus improve the detection of fluorescence.

[0059] In one embodiment, the housing is formed of one or more components that are attached to each other by attachment means.

[0060] In one embodiment, the housing is formed of two parts that are attached to each other by attachment means. In a preferred embodiment, the attachment means are each independently selected from the group consisting of nails, clips, vacuum suction cups, screws, hooks, combinations of bolts and nuts, brackets, locks, tapes, snap locks, hinges, seals, and one or more magnets. It should be understood that the attachment means are for attaching two or more parts forming the housing and / or for at least partially surrounding the periphery of the transparent conduit and / or for attaching to the outside of the conduit.

[0061] In one embodiment, the computing device is connected to and preferably communicatively coupled to at least one of one or more light sources and / or at least one of one or more light receivers. Preferably, the computing device is communicatively coupled to at least one of one or more light sources and / or at least one of one or more light receivers by wired connection means or wireless connection means. In a preferred embodiment, the computing device is communicatively coupled to one or more light receivers provided within the housing.

[0062] In a preferred embodiment, the housing - one or more light sources capable of operating to emit light towards a sample carried by a transparent conduit, - one or more light receivers capable of operating to detect fluorescence generated by one or more fluorophores in a sample carried by the transparent conduit, - the computing device is communicatively coupled to at least one of one or more light sources, and the computing device - receives a first signal from at least one of one or more light receivers, the first signal being proportional to the fluorescence detected by at least one of one or more light receivers, - compares the first signal with a predetermined threshold value for the first signal, and - It has computing means for outputting first information representing a comparison result.

[0063] Therefore, it should be understood that the computing device of the present apparatus is provided within the housing of the present apparatus.

[0064] In one embodiment, the housing further has one or more seals that adhere the housing to a transparent conduit. The seals may also reduce or prevent light from the outside from entering the housing.

[0065] In a preferred embodiment, the housing is provided at longitudinal ends of the housing on opposite sides of each other, and further has one or more seals that adhere the housing to a transparent conduit. When surrounding the conduit, the seal may be an adjustable seal that allows the housing to be adhered to transparent conduits of various sizes, for example, when the housing surrounds a conduit having an outer diameter of 1 mm (3 Fr) to 20 mm (60 Fr), preferably a surgical suction tube or a medical tube having an outer diameter of 2.67 mm (8 Fr) to 16 mm (48 Fr). The housing may have, for example, 1, 2, 3, 4, 5, 6, 7, or 8 seals.

[0066] In one embodiment, one or more light sources are operable to emit light at one or more fluorescence excitation curve wavelengths of one or more fluorophores in the sample, independently of each other. In a preferred embodiment, one or more light sources are operable to emit light at the peak wavelength of the fluorescence excitation curve of one or more fluorophores contained in the sample, or in a wavelength range including the peak wavelength of the fluorescence excitation curve, independently of each other. In a preferred embodiment, one or more light sources are operable to emit light at one or more wavelengths independently selected from the group consisting of wavelengths in the range of 350 nm to 430 nm, 600 nm to 700 nm, and / or 750 nm to 850 nm, independently of each other. In a preferred embodiment, one or more light sources are selected from the group consisting of PpIX, indocyanine green, methylene blue, fluorescein, and salts thereof, such as cyanine, T700, T800, BLZ-100, GB119, IRDye800CW conjugate, IRDye700Dx conjugate, EC17, LUM015, AVB-620, folic acid-fluorescein isothiocyanate (FITC), OTL38, gGlu-HMRG, green fluorophore conjugate, fluorescent dye-labeled peptide, fluorophore-conjugated antibody, fluorescent nanoparticles, activatable fluorescent probes, endogenous fluorophores, fluorescent bacteria, fluorescent viruses, NAP(P)H, FAD, flavin, collagen, vitamins such as vitamin A1, vitamin B2, vitamin B6, and vitamin B9, indoleamine, mitochondria and lysosomes, and derivatives, isomers and salts thereof, or combinations thereof, preferably operable to emit light at the wavelength of the fluorescence excitation curve of a fluorophore selected from PpIX and its derivatives, indocyanine green, methylene blue, fluorescein, and salts thereof. In a preferred embodiment, one or more light sources are operable to emit light at a wavelength of about 405 nm and / or about 633 nm, independently of each other. The housing has one, two, three, four or more light sources operable to emit light at one or more fluorescence excitation curve wavelengths of one or more fluorophores in the sample, independently of each other.

[0067] In one embodiment, one or more light receivers are operable to detect fluorescence of one or more wavelengths of one or more fluorescence emission curves of one or more fluorophores in a sample, independently of each other. In a preferred embodiment, one or more light receivers are operable to detect fluorescence of the peak fluorescence emission wavelength of one or more fluorophores contained in a sample, independently of each other. In a preferred embodiment, one or more light receivers are operable to detect fluorescence of one or more wavelengths independently selected from the group consisting of wavelengths from 600 nm to 655 nm, wavelengths from 650 nm to 695 nm, wavelengths from 700 nm to 790 nm, and / or wavelengths from 790 nm to 840 nm, independently of each other. In a preferred embodiment, one or more light receivers are selected from the group consisting of PpIX, indocyanine green, methylene blue, fluorescein, and salts thereof, such as cyanine, T700, T800, BLZ-100, GB119, IRDye800CW conjugate, IRDye700Dx conjugate, EC17, LUM015, AVB-620, folic acid-fluorescein isothiocyanate (FITC), OTL38, gGlu-HMRG, green fluorophore conjugate, fluorescent dye-labeled peptide, fluorophore-conjugated antibody, fluorescent nanoparticles, activatable fluorescent probe, endogenous fluorophore, fluorescent bacteria, fluorescent virus, NAP(P)H, FAD, flavin, collagen, vitamins such as vitamin A1, vitamin B2, vitamin B6, and vitamin B9, indoleamine, mitochondria and lysosomes, and derivatives, isomers and salts thereof, or combinations thereof, preferably operable to detect fluorescence resulting from one or more fluorophores selected from the group consisting of PpIX and derivatives thereof, indocyanine green, methylene blue, fluorescein, and salts thereof. The housing has one, two, three, four or more light sources operable to detect fluorescence of one or more wavelengths of one or more fluorescence emission curves of one or more fluorophores contained in a sample, independently of each other. Fluorescence generated by one or more fluorophores may vary depending on factors such as, for example, the pH and temperature of the sample.

[0068] In a preferred embodiment, one or more light sources are operable to emit light at the peak wavelength of the fluorescence excitation curve of PpIX or a derivative thereof, and one or more light receivers are operable to detect fluorescence generated by PpIX or a derivative thereof. It should be understood that a sample containing PpIX may represent cancer, and the PpIX in the sample may be derived from 5-ALA or its prodrug, or another drug orally administered to the subject, for example, 1 to 24 hours before obtaining the sample.

[0069] Additionally or alternatively, each of the one or more light sources further has one or more emission light filters and / or each of the light receivers has one or more incident light filters. In a preferred embodiment, each of the one or more emission light filters is separately and independently selected based on one or more peak wavelengths of the fluorescence excitation curves of the light source and one or more fluorophores, and each of the one or more incident light filters is separately and independently selected based on one or more peak fluorescence emission wavelengths of the light receiver and one or more fluorophores.

[0070] As used herein and hereinafter, the term "light filter" means a device that selectively transmits light of different desired wavelengths, and is embodied, for example, as a glass surface or a plastic device in the optical path. The optical properties of a light filter are described by these frequency responses, and such frequency responses specify how the amplitude and phase of each frequency component of the incident light are changed by the filter. Examples of light filters include, but are not limited to, absorption filters, dichroic filters, monochromatic filters, infrared transmission filters, infrared cut-off filters, band-stop filters, long-pass filters, band-pass filters, short-pass filters, and ultraviolet filters. It should be understood that the light filters are preferably emission light filters and incident light filters, that is, the emission light filters are used to change the wavelength and / or intensity of the light emitted by the light source, and the incident light filters are used to change the wavelength and / or intensity of the light received by the light receiver.

[0071] In one embodiment, each of one or more fluorophores in the sample is separately and independently selected from the group consisting of PpIX, indocyanine green, methylene blue, fluorescein, and salts thereof, cyanine, T700, T800, BLZ-100, GB119, IRDye800CW conjugate, IRDye700Dx conjugate, EC17, LUM015, AVB-620, folic acid-fluorescein isothiocyanate (FITC), OTL38, gGlu-HMRG, green fluorophore conjugate, fluorescent dye-labeled peptide, fluorophore-conjugated antibody, fluorescent nanoparticles, activatable fluorescent probe, endogenous fluorophore, or combinations thereof.

[0072] In one embodiment, at least one of the one or more light sources is arranged in front of the one or more light receivers with respect to the transport direction of the sample carried in the transparent conduit. By arranging at least one of the one or more light sources and the one or more light receivers in this way, more effective excitation of the one or more fluorophores is possible.

[0073] Additionally or alternatively, the housing has at least a first light source, the one or more light receivers have at least a first light receiver and a second light receiver, these light receivers are positioned such that the distance from the first light receiver to the first light source is shorter than the distance from the second light receiver to the first light source, and the computing means further - receives a first signal from the first light receiver, the first signal being proportional to the fluorescence detected by the first light receiver, - determines a verification value for a second signal in response to receiving the first signal, - receives a second signal from the second light receiver, the second signal being proportional to the fluorescence detected by the second light receiver, - compares the second signal with the verification value, - outputs first information regarding the second signal in response to the second signal exceeding or being equal to the verification value, and - In response to the second signal not exceeding or being unequal to the verification value, verifying the detection of fluorescence from two or more light receivers by a first light receiver by outputting second information regarding the second signal.

[0074] It should be understood that different fluorophores may have different fluorescence induction rates and fluorescence decays. Due to the excitation light, the fluorescence of the fluorophore can be induced after a specific time, that is, induced by the fluorescence induction rate. Fluorescence decay means the time when the fluorophore emits fluorescence after being excited by the excitation light. When receiving the first signal, the computing means takes into account the distance between the first light source, the first light receiver and the second light receiver, and one or more of the fluorescence induction rate and / or fluorescence decay of the fluorophore, and can determine what the value of the second signal should be (the verification value of the second signal). When the second signal received by the second light receiver exceeds or is equal to the verification value, the first information output of the second signal may be different from the second information output because the second signal received by the second light receiver does not exceed or is not equal to the verification value.

[0075] By using at least two light receivers, the sensitivity and / or specificity of fluorescence detection can be improved.

[0076] Additionally or alternatively, the housing includes at least the first light source, the one or more light receivers include at least the first light receiver and the second light receiver, and the distance between at least the first light receiver and the second light receiver is selected from one or more distances based on the fluorescence induction rate, fluorescence decay, and fluorescence intensity of a sample containing one or more fluorophores, or a combination thereof.

[0077] It should be understood that the locations of the one or more light sources and the one or more light receivers may be at different locations of the housing. Depending on the fluorescence induction rate, fluorescence decay, and fluorescence intensity of the fluorophore in the sample, it is advisable to select the distance between the first light receiver and the second light receiver. For example, when the fluorescence induction rate is low (short) and / or the fluorescence decay is long, this distance should be longer than when the induction rate is high (long) and / or the fluorescence decay is short. When the fluorescence intensity is high, this distance should be long because the fluorescence can be detected over a long period of time. Additionally or alternatively, the computing means for calculating the flow rate of the sample carried within the transparent conduit may be provided within a computing device. To calculate the flow rate, divide the distance from the first light receiver to the second light receiver by the time difference between the first signal received from the first light receiver and the second signal received from the second light receiver. The calculated flow rate can be used for calibration of the device.

[0078] In one embodiment, the housing surrounds the transparent conduit at a distance of 0.01 to 10 m, preferably 0.3 to 10 m from the object from which the sample is obtained, more preferably 0.3 to 3 m from the object, and even more preferably 0.3 to 1 m from the object. The distance between the housing and the object is not limited as long as the sample is being carried through the transparent conduit. This allows the device to be used, for example, during surgery. When near real-time detection is required, the distance from the housing to the object should preferably be short enough to allow for near real-time feedback to the user of the device, and the housing should not significantly and negatively interfere with the user's work. The housing may preferably be configured to be removable so as to surround at least a portion of the transparent conduit. Thus, one advantage of the present device is that it can be removed from the transparent conduit when not in use, and the distance from the housing to the object can be selected according to how much space there is around the object.

[0079] In one embodiment, the computing device is provided within the housing.

[0080] In one embodiment, at least one processor, a memory, computer program code, and a user interface constitute the computing means of a computing device.

[0081] In one embodiment, the housing further has an ON / OFF switch and means for outputting information, such as a user interface, an augmented reality interface, acoustic means, a display, and an LED. Additionally, the housing may further have at least one of an LED indicating the state of the device, the success or failure of calibration, reliable detection, and / or the battery level, as well as a charging port and / or a data port.

[0082] In one embodiment, a computing device is communicably coupled to at least one of one or more light receivers, and the computing device - has at least one processor, and - has at least one memory including computer program code, and the at least one memory and the computer program code cause the at least one processor to - receive a first signal from at least one of one or more light receivers, the first signal being proportional to fluorescence detected by at least one of one or more light receivers, - compare the first signal with a predetermined threshold value regarding the first signal, and - perform an operation of outputting first information representing a comparison result.

[0083] In another aspect, an embodiment of the present invention is a method for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores and representing a disease or a pathological condition, the sample being carried out from the subject through a transparent conduit, and the method includes i) a step of carrying out a sample containing one or more fluorophores and representing a disease or a pathological condition from the subject through a transparent conduit, and ii) emitting light towards the sample being carried by the conduit; iii) detecting fluorescence generated by one or more fluorophores in the sample being carried by the conduit; iv) generating a first signal proportional to the detected fluorescence; v) comparing the first signal with a predetermined threshold value for the first signal; vi) outputting first information representing the comparison result. A method is provided which is characterized by including the above steps.

[0084] In one embodiment, the method further includes, before step i) or ii), preferably before step i), configuring the housing to surround at least a part of the conduit that is transparent, and the housing has - one or more light sources capable of operating to emit light towards the sample being carried by the transparent conduit; - one or more light receivers capable of operating to detect fluorescence generated by one or more fluorophores in the sample being carried by the transparent conduit, and further optionally - a computing device including computing means for performing steps v) and vi).

[0085] Additionally or alternatively, the method described in the present disclosure is a method for detecting the fluorescence of a sample in substantially real time.

[0086] In one embodiment, the sample is carried away from the subject by negative pressure. It should be understood that negative pressure is a pressure lower than the ambient pressure. Negative pressure can be achieved, for example, by an ultrasonic aspirator or a surgical suction device.

[0087] In one embodiment, the method further includes obtaining a sample and emitting light towards the subject before the sample is carried away from the subject within the conduit. Preferably, the light excites one or more fluorophores in the sample.

[0088] In one embodiment, the method further includes a calibration step, and the sample is a calibration composition containing fluorophores of known fluorescence and concentration. Preferably, the calibration step is performed before step i).

[0089] Additionally or alternatively, the method further includes a step of verifying fluorescence detection, and the step of verifying fluorescence detection includes - detecting a second fluorescence generated by one or more fluorophores in the sample being carried in the conduit; - generating a second signal proportional to the detected second fluorescence; - determining a verification value for the second signal in response to the generated first signal; - comparing the second signal with the verification value; - outputting second information regarding the second signal in response to the second signal exceeding or being equal to the verification value; and - outputting third information regarding the second signal in response to the second signal not exceeding or being equal to the verification value.

[0090] Additionally or alternatively, the method further includes a step of determining the flow rate, the sample is a calibration composition containing fluorophores of known fluorescence and concentration, and the step of determining the flow rate includes - calculating a first time value based on the time difference between the step of generating the first signal and the step of generating the second signal in response to the generation of the first signal and the second signal; - calculating a flow rate value based on the result of dividing a predetermined distance value by the first time value in response to the calculation of the first time value; - outputting fifth information indicating the calculation result of the flow rate value.

[0091] In one embodiment of the present invention, a method for detecting the fluorescence of a sample obtained from a subject, the sample contains one or more fluorophores, represents a disease or pathological condition, the sample is carried out from the subject by a transparent conduit, and the method includes i) a step of taking out a sample, preferably a part of a tumor, from a subject, preferably from a human body; ii) a step of transporting a sample containing one or more fluorophores and representing a disease or a pathological condition out of the subject in a transparent conduit; iii) a step of emitting light toward the sample being transported in the conduit; iv) a step of detecting fluorescence generated by one or more fluorophores in the sample being transported in the conduit; v) a step of generating a first signal proportional to the detected fluorescence; vi) a step of comparing the first signal with a predetermined threshold value for the first signal; vii) a method is provided that includes a step of outputting first information representing the comparison result.

[0092] In another aspect, an embodiment of the present invention provides the use of the device disclosed herein for carrying out a method selected from the group consisting of an invasive medical method, a method for detecting diseased tissue or diseased body fluid containing one or more fluorophores and representing a disease or a pathological condition, a method for detecting a tumor, and a diagnosis of cancer. Preferably, the diagnosis of cancer is an in vitro or in vitro diagnosis of cancer.

[0093] From another perspective, one embodiment of the present invention provides a component kit in which the device described in the present disclosure is combined with a transparent conduit. In one embodiment, the periphery of the conduit is such that the housing can surround the entire periphery of the conduit. In one embodiment, the outer diameter of the conduit can be that of a surgical suction tube or a medical tube with an outer diameter of 1 mm (3 Fr) to 60 mm (180 Fr), preferably 1 mm (3 Fr) to 20 mm (60 Fr), more preferably 1 mm (3 Fr) to 16 mm (48 Fr), still more preferably 2 mm (6 Fr) to 16 mm (48 Fr), and even more preferably 2.67 mm (8 Fr) to 16 mm (48 Fr). The inner diameter of the conduit with such an outer diameter can be an inner diameter that can carry a sample within the conduit and that can be used for medical procedures. For example, the inner diameter of the conduit is preferably 0.75 mm to 15.75 mm. In one embodiment, the component kit further includes a leaflet with information on how to use the present device.

[0094] From another perspective, one embodiment of the present invention provides a surgical method, a method for treating a disease or a pathological condition, or an in vivo diagnosis of a disease or a pathological condition using the device disclosed in this specification. In one embodiment, the disease is cancer. In one embodiment, the surgical method includes cancer surgery.

[0095] From another perspective, one embodiment of the present invention provides a method for detecting the fluorescence of a sample containing one or more fluorophores and representing a disease or a pathological condition using the device disclosed in this specification.

[0096] From another perspective, one embodiment of the present invention provides the device disclosed in this specification for implementing the method disclosed in this specification.

[0097] Experimental section

[0098] In an exemplary embodiment, experiments were conducted at a microsurgical (ultramicroscopic surgery) research and training center. The experiments describe the characteristics of the method for detecting fluorescence of a sample as described in the present disclosure. The sample contains PpIX at a concentration representing active glioma, and the sample can be detected as well as or better than those skilled in the art either as a good sample or by the fluorescence of PpIX. The fluorescence of a sample containing a fluorophore, i.e., a sample being carried away from a subject in a transparent conduit, was detected from the surgical resection fluid flowing through a suction tube.

[0099] The homologous PpIX samples were prepared according to the in vivo PpIX concentration ranges within grade IV gliomas recorded during resection as described by Johansson et al. (Johansson, Ann, et al., “5-Aminolevulinic acid-induced protoporphyrin IX levels in tissue of human malignant brain tumor”, Photochemistry and Photobiology, 2010, 86.6, pp. 1373-1378). The PpIX samples represented a disease or pathological condition. The samples were aspirated (i.e., carried away) by a surgical suction pump system and the fluorescence was detected with the apparatus disclosed herein. Liquids mimicking placental tissue debris and blood were aspirated together with the PpIX samples and also separately between the PpIX samples to simulate the surgery during resection of active glioma cells and surrounding healthy nerve tissue.

[0100] The fluorescent PpIX solution was prepared in a histochemistry laboratory according to Taniguchi, Hiroki, et al. 「Improving convenience and reliability of 5-ALA-induced fluorescent imaging for brain tumor surgery」, International Conference on Medical Image Computing and Computer-Assisted Intervention, Springer, Cham, 2015. Physiological saline (B. Braun Melsungen AG, Germany) was heated to +100 °C, gelatin (#G2500, Sigma-Aldrich Company, St. Louis, Missouri, USA) was added, and the mixture was allowed to cool to +37 °C. The disodium salt of PpIX (#258385, Sigma-Aldrich, 2.02 mg) was dissolved in dimethyl sulfoxide (DMSO, #D4540, Sigma-Aldrich, 20 mL) to produce a PpIX solution. To obtain PpIX samples at a predetermined PpIX concentration, 1.33 mL of the PpIX solution or DMSO (1.33 mL) was mixed with 34.7 mL of the gelatin-saline mixture, and then 4.0 mL of Intralipid solution 20% (a fat emulsion containing 20% (w / v) soybean oil, egg lecithin, glycerol, sodium hydroxide, and water, Fresenius Kabi AB, Uppsala, Sweden) was added. The resulting gel-like PpIX samples were solidified and stored at a constant temperature (+4 °C).The PpIX sample was prepared according to the following equation (1), where in the following formula, D = the concentration (mol / l) of PpIX as the free acid in the PpIX sample, C ≒ 3.0×10. -6 mol / l, and k = 0 or 2.00. D = kC (1)

[0101] The PpIX samples were classified into two concentration groups. The first concentration group included three 2C PpIX samples, and the second concentration group included eight control samples (D = 0C) according to visual detection by humans (enclosed in parentheses) determined by a skilled surgeon. 1.0C PpIX (control sample, no PpIX, no fluorescence) 2.2C PpIX+++ (true positive sample, about 6.0×10 -6 mol / L PpIX, representing grade IV glioma cells with activity, and moreover, fluorescence visible to experts)

[0102] A 3D printed test platform including hexagonal slots (1 cm in diameter) and matching lids to protect the samples from light exposure was manufactured using polylactic acid (PLA) by a Zmorph 2.0SX - 3D printer. The platform was designed in Autodesk Fusion 360 (v.2.0.6037) and manufactured such that the aspiration of the sample could be performed quickly as in the case of glioma surgery. Thus, the slot represents the subject matter of the present disclosure from which the sample is obtained.

[0103] The surgical suction pump (Medela Dominant Flex) was set to an effective constant maximum vacuum of -80 kPa equivalent to a flow rate of 50 L / min. A suction handle (Mediplast 6066500400BP) (4.0 mm in diameter, 80 mm in length) made of stainless steel and equipped with a round hub was attached to a transparent sterile PVC suction tube (CH25, 5.8 / 8.3 mm in diameter, 3.5 m in length). The other end of the tube was connected to a 1-liter suction waste container. The same suction tube and suction waste container were used within the operating room environment. These samples were aspirated via a metal suction handle to mimic the size and composition of the tumor tissue from which the samples were taken. The device for detecting fluorescence surrounded the PVC suction tube at a point 1 / 3 of the length of the PCV suction tube from the suction handle, i.e., at about 1.17 m from the suction handle and about 1.25 m (i.e., about 1.25 m from the subject) from the tip of the suction handle.

[0104] A blood mimic solution was prepared by adding a red dye (Dr. Oet-ker Red coloring agent) to water at a ratio of 1:20 (e.g., 5 mL of red dye and 100 mL of water). Small pieces of biological tissue debris were added to the PpIX sample before aspiration to add noise to the fluorescence detection and to verify the reliability of the method. Since the analysis was performed in a room designed for surgery and spectral experiments, background lighting could be eliminated.

[0105] A narrow-band LED (M405l2 UV (405 nm) - mounted LED, 1000 mA, 410 mW (min), manufactured by Thorlabs) was used as a fluorescence excitation source, and light was emitted toward the sample being transported in the conduit. Fluorescence generated by the fluorophore in the sample being transported in the conduit was detected using a Hamamatsu Photonics PMA-11 spectrometer (model c7473-36, connected to a light receiver and a computer (computing device)). The light source and the light receiver were arranged perpendicular to the conduit, that is, the angle formed by the longitudinal central axis of the light source and the longitudinal central axis of the light receiver was approximately 90°. The excitation light source induced fluorescence of the PpIX sample in the surgical drainage fluid, and the fluorescence was detected by the spectrometer almost in real time. Experiments were conducted using narrow-band pass filters for excitation and emission light. Blue light from the excitation light source was filtered by using a Semrock 414 / 46-Brightline single-band filter to eliminate light other than the desired excitation light. A Semrock 632 / 22-Brightline single-band filter (incoming light filter) was attached to the spectrometer to eliminate background illumination.

[0106] Detection and analysis were carried out for both 2C PpIX samples and 0C PpIX samples using the same suction rate (50 L / min) and spectrometer exposure time (80 ms) in three different test execution variations, that is, i) a PpIX sample (2C or 0C) + water was placed in the slot, ii) a PpIX sample (2C or 0C) + blood mimic fluid was placed in the slot, or iii) a PpIX sample (2C or 0C) + blood mimic fluid + debris of placental tissue was placed in the slot. Every second slot was washed at the detection site and filled with a control fluid (water) that mimicked the physiological saline solution aspirated during glioma surgery. The spectrometer was set to perform 100 fluorescence detection measurements, and such measurements took 8 seconds with an exposure time of 80 ms. The exposure time corresponded to the length of time the detector was exposed to light.

[0107] Two PpIX sample concentrations (2C and 0C) were used, and all three test execution variations were performed without a PpIX sample representing background noise. Before the test executions, the 2C and 0C PpIX samples were cut into cube-shaped pieces 2 - 4 mm in length. Similarly, pieces of placental tissue were cut into cube-shaped pieces 2 - 4 mm in length. i) In the first test execution, three pieces of the 2C PpIX sample were placed in three slots together with 1 mL of water, and three pieces of the 0C PpIX sample were placed in three slots together with 1 mL of water, i.e., all six slots in total contained three pieces each of either the 2C PpIX sample or the 0C PpIX sample. ii) In the second test execution, three evenly prepared bits were placed in three slots together with 1 mL of blood mimicking fluid, as in the case of the first test execution. iii) Regarding the third test execution, three equally prepared bits were placed in all three together with 1 mL of blood mimicking fluid and three pieces of placental tissue per slot, as in the case of the first test execution, to test whether the randomly aspirated remaining tissue would affect fluorescence detection and analysis by the spectrometer. As a result, in each test execution variation analysis, a total of nine pieces, both 2C and 0C, were obtained.

[0108] Similar to the case of sample measurement, background noise was determined using the same aspiration rate and exposure time, and the three test execution variations were performed without the PpIX sample. The background noise was recorded as the number of photon counts recorded by the spectrometer.

[0109] During the fluorescence detection of the PpIX sample, the PpIX sample was regarded as detected when the fluorescence at 630 nm exceeded a predetermined threshold. In this method, the predetermined threshold was determined to be the level obtained by adding 40% of the average background noise level to the average background noise level. The average background noise level was equal to 560 photon counts (per 80 ms), and thus the predetermined threshold was 784 photon counts. The photon counts corresponded to the photocurrent (signal) received from the spectrometer's photoreceptor by the computing means of the computing device.

[0110] For each test execution variation, the number of fluorescence signal peaks where the photon counts (photocurrent proportional to the fluorescence detected by the photoreceptor), corresponding to the number of detected PpIX samples, exceeded the predetermined threshold was counted. Additionally, the average value of the photon counts (i.e., the average value of the maximum photocurrent) at each fluorescence signal peak (light at a wavelength of 630 nm) where the photon counts exceeded the specified threshold was calculated. Thus, signals proportional to the detected fluorescence were generated and compared with the predetermined threshold. Table 1 lists the average value of the fluorescence intensity and the number of detected fluorescence intensity peaks for each test execution variation performed. TIFF0007690040000001.tif61168

[0111] Table 1. Summary of measurement results. 1 Test execution runs 1 to 3 each contain nine 2C PpIX samples, and test execution variations 4 to 6 are each composed of nine 0C PpIX samples (2C ≈ 6.0 μM, 0C = no PpIX), placenta = placental tissue debris. 2 The number of fluorescence signal peaks where the photon counts exceeded the predetermined threshold. 3 The calculated average photon count at each fluorescence signal peak where the photon counts exceeded the predetermined threshold (at 630 nm) with an exposure time of 80 ms. The predetermined threshold for positive detected peaks was set at 784 photon counts (background noise × 1.4).

[0112] According to the verification of the experimental results, when the fluorescence of the 2C PpIX sample, that is, the photon count (fluorescence) observed in the sample mimicking grade IV glioma cells exceeded the photon count of the control sample and exceeded the preset threshold by 3 to 4 times, that is, when the information indicating the comparison result was output, it was demonstrated that it was detected with high reliability. Fluorescence and 2C samples were also detected in the presence of blood mimic fluid (red dye + water) and / or other biological samples (placental tissue debris) representing random surgical aspiration debris. Of all 27 2C PpIX samples, 18 were detected separately and 9 could not be detected. Therefore, at least 66% of the maximum number of 2C PpIX samples were detected. The samples were of various sizes (2 - 4 mm), and since the photodetector might have detected the fluorescence of multiple samples simultaneously, there might have been 2C PpIX samples that were not detected among the detected samples. One false positive count of 0C was detected, but only 28 photon counts exceeded the preset threshold. Since all photon counts of the detected 2C PpIX samples were significantly higher than the photon counts of the 0C samples and background noise, the false positive count could be safely excluded by increasing the preset threshold to, for example, 813 photon counts.

[0113] Nearly real - time detection of fluorophores in samples being carried in a transparent conduit was achieved in about 0.04 seconds and calculated according to the following equation (2). In the following formula, Q = flow rate (m 3 / s), V 1 = aspiration handle volume (m 3 ), V 2 = cylinder volume (m 3 ), A 1 = aspiration handle area (m 2 ), A 2 = cylinder area (m 2 ), d 1 = aspiration handle length (m), d 2 = cylinder length (m), r 1 = aspiration handle radius (m), r2 = the cylinder radius (m). JPEG0007690040000002.jpg15153 JPEG0007690040000003.jpg31170

[0114] The usefulness of the method described in the present disclosure was successfully demonstrated, that is, the fluorescence of one or more wavelengths of one or more fluorescence emission curves of one or more fluorophores in a sample was detected according to the method, and the sample was contained in a fast-flowing fluid carried in a transparent conduit in a novel manner not previously described. The results of the fluorescence detection method were output almost in real time together with information indicating whether, for example, the photon count (the photocurrent proportional to the detected fluorescence) exceeded, was equal to, or did not exceed or was equal to a predetermined threshold. According to the demonstration of the test results, the method of the present invention can be used in clinical practice. Regarding the sensitivity to detect clinically relevant concentrations of PpIX, the present invention was also able to perform the output of information on accurate fluorescence intensity (photon count of the signal peak) that cannot be performed by experts.

[0115] FIG. 1 is a block diagram of an apparatus 100 for detecting the fluorescence of a sample obtained from a subject according to an embodiment of the present invention. The sample contains one or more fluorophores and represents a disease or pathological condition, and the sample is carried out from the subject through a transparent conduit. The apparatus 100 has a housing 101, a computing device 180, one or more light sources 110, and one or more light receivers 120. The computing device 180 may be connected to at least one of the one or more light receivers within the housing 101. The connection can be realized by wire connection means or wireless connection means. In one embodiment, the computing device 180 may be connected to the housing 101. The computing device 180 may be provided inside the housing 101 or outside the housing 101.

[0116] FIG. 2 shows an embodiment of an apparatus 100 for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores and representing a disease or medical condition, the sample being conveyed from the subject through a transparent conduit. The apparatus 100 includes a housing 101 having one or more light sources (not shown) and one or more light receivers (not shown), and has a computing device (the computing device is not shown). The housing 101 may be configured to surround at least a portion of the transparent conduit 131, or the housing 101 may be configured to completely surround the transparent conduit 131. The housing 101 preferably surrounds a portion or the entire perimeter of a transparent conduit 131 that is preferably attached to suction unit attachment means 141 for a suction unit, the transparent conduit 131 is preferably attached to a tube connector 161, and the tube connector 161 is preferably attached to a suction tip 151. The suction unit may be operable to create a negative pressure for removing and / or transporting the sample from the subject (the suction unit and the subject are not shown). The one or more light sources and the one or more light receivers provided within the housing 101 are not shown. The computing device (not shown) may be connected to at least one of the one or more light receivers provided within the housing 101. The connection may be realized by wire connection means 190 or by wireless connection means.

[0117] Figure 3 shows an embodiment of an apparatus 100 for detecting fluorescence of a sample obtained from a subject, where the sample contains one or more fluorophores and represents a disease or medical condition, and the sample is carried out from the subject by a transparent conduit. The apparatus 100 has a housing 101 and a computing device (the computing device is not shown). The housing 101 preferably has one or more light sources 110 and one or more light receivers 120. The computing device (not shown) may be connected to the housing 101. The connection to at least one of the one or more light receivers 120 may be realized by wire connection means 190 or by wireless connection means. The computing device 180 may be provided inside the housing 101 or outside the housing 101. This figure shows a cross-section of the transparent conduit 131. The housing 101 preferably surrounds at least a part of the conduit 131. The housing 101 may surround the entire periphery of the transparent conduit 131. Light 191 may be emitted by the light source 110 from a subject (not shown) and directed towards the samples 118, 119 carried out from the subject. The sample 118 preferably contains one or more fluorophores and represents a disease or medical condition. The sample 119 may not be fluorescent or may not contain one or more fluorophores. A filter 115 for emitted light may be used in combination with the light source 110 to filter the emitted light. The wavelength of the light emitted by the one or more light sources and the wavelength of the light filtered by the emitted light filter 115 may be different from each other. The emitted light and the light filtered by the emitted light filter 115 are indicated by the dashed arrow 191 in Figure 3. The one or more light sources 110 may be operable to emit light 191 towards the conduit 131 and the samples 118, 119 within the conduit 131 while the samples 118, 119 are being carried within the conduit 131. The excited fluorophores in the sample 118 can emit fluorescence 192. The fluorescence 192 may be detected by the light receiver 120. The wavelengths of the emitted light 191 and the fluorescence 192 may be different from each other.The filter 125 for incident light may be used in a state of being coupled to the light receiver 120 to filter the wavelength of the fluorescence 192 emitted. The wavelength of the fluorescence emitted by the sample 118 and the wavelength of the fluorescence filtered by the filter 125 for incident light may be different from each other. The fluorescence and the fluorescence after filtering are indicated by the arrow 192 in FIG. 3. The transport direction (i.e., the flow direction) of the samples 118, 119 from an object (not shown) in the transparent conduit 131 is indicated by the thick arrow 132. As shown in FIG. 3, at least one of the one or more light sources 110 may be arranged in front of the one or more light receivers 120 when viewed in the transport direction of the sample being conveyed in the transparent conduit 131.

[0118] Figure 4 shows an embodiment of an apparatus 100 for detecting fluorescence of a sample obtained from a subject, where the sample contains one or more fluorophores, represents a disease or medical condition, and the sample is carried out from the subject through a transparent conduit. The apparatus 100 has a housing 101 and a computing device 180, and the computing device 180 may be provided inside the housing 101 or outside the housing 101. The housing 101 further has one or more light sources 110 and one or more light receivers 120, 121 therein. The computing device 180 may be communicatively coupled to at least one of the one or more light receivers 120, 121 and / or at least one of the one or more light sources 110. The coupling may be realized by wire connection means or by wireless connection means (not shown). The one or more light sources 110 may further have a filter 115 for emitted light, and each of the one or more light receivers 120, 121 may further have a filter 125 for incident light. The housing 101 may be formed of one or more components. The housing 101 attaches (e.g., includes a hinge as attachment means 103) and / or locks (e.g., includes magnets as attachment means 104a, 104b, 105a, 105b) one or more components that can form the housing 101 and / or may include attachment means 103, 104a, 104b, 105a, 105b for attaching to a conduit (not shown) from the outside. It should be understood that the housing 101 may be removably configured to surround at least a portion of the transparent conduit or may surround the entire perimeter of a transparent conduit (not shown). Thus, the housing 101 may be in an "open position", a "closed position", or a position between an "open position" and a "closed position". In the "closed position", the housing 101 may surround the entire perimeter of the transparent conduit, and the attachment means 104a, 104b, 105a, 105b may be arranged to lock one or more components in the "closed position".Furthermore, the housing 101 preferably has openings 106a to 106d at longitudinal ends of the housing on opposite sides of each other. The housing 101 preferably has one or more seals 107a to 107d for adhering the housing to a transparent conduit (not shown). The seal may be an adjustable seal. Furthermore, the housing 101 preferably has a power supply device 108. The power supply device preferably has one or more components 108a to 108h, and each component 108a to 108h may be the same or different from each other, and each component is preferably connected (electrically coupled) to one or more light sources 110, one or more light receivers 120, 121 and / or a computing device 180 (connection means are not shown). Furthermore, the housing 101 preferably has one or more display marks 109a to 109f for indicating the mounting direction of the housing 101 around the conduit. The one or more display marks 109a to 109f can indicate the transport direction of a sample being carried out from an object in the conduit (the sample, the object, and the conduit are not shown). At least one of the one or more light sources 110 is preferably arranged in front of the one or more light receivers 120, 121 when viewed in the transport direction of the sample being carried out in the transparent conduit.

[0119] FIG. 5 shows an embodiment of an apparatus 100 for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores and representing a disease or a medical condition, the sample being carried out from the subject by a transparent conduit. The apparatus 100 has a computing device (not shown) and a housing (not shown), and the housing preferably surrounds at least a portion of the transparent conduit 131. The housing has one or more light sources 110 and one or more light receivers 120. The computing device (not shown) may be communicatively coupled to at least one of the one or more light receivers 120. A sample 118 containing one or more fluorophores and representing a disease or a medical condition and a sample 119 that is non-fluorescent or does not contain one or more fluorophores are shown. Further, this figure shows a cross-section of the transparent conduit 131, and the housing 101 surrounds at least a portion of this transparent conduit. The angle 134 formed by the longitudinal central axis 133 of at least one of the one or more light sources 110 and the longitudinal central axis 135 of at least one of the one or more light receivers 120 may be in the range from 0° to 180°. In one embodiment, the angle 134 may be in the range from 15° to 180°. In one embodiment, this angle may be within the range from 90° to 180°, preferably 90°.

[0120] FIG. 6 shows an example of representing the photocurrent I (mA) (corresponding to one or more signals indicated by the black line) from at least one of the one or more light receivers 120 as a function of the time t (ms) received by the computing device 180. A predetermined threshold value A1 of the photocurrent (signal) is indicated by the dotted line. The fluorescence 192 generated by one or more fluorophores in the sample 118 may be detected by one or more light receivers 120. The computing device 180, which may be communicatively coupled to at least one of the one or more light receivers 120, may receive the signal from the light receiver 120. This signal may correspond to the photocurrent I and may be proportional to the fluorescence detected by at least one of the one or more light receivers 120. The computing device 180 may compare this signal (photocurrent) with a predetermined threshold value A1 for the signal (photocurrent). Also shown are two signal peaks P1 (photocurrent I1 at time t1) and P2 (photocurrent I2 at time t2) corresponding to the maximum photocurrent. It may be proportional to the maximum fluorescence generated by one or more fluorophores in two samples 118a, 118b detected by at least one of the one or more light receivers 120. The samples 118a, 118b are obtained from the subject and carried from the subject by the transparent conduit 131. The two fluorescent samples 118a, 118b passed through at least one of the one or more light receivers 120 at two different time points (t1, t2). P1 may correspond to the maximum fluorescence generated by the first sample 118a that passed through at least one of the one or more light receivers 120 at time t1, and P2 may correspond to the maximum fluorescence generated by the second sample 118b that passes through at least one of the one or more light receivers 120 at time t2. Both signal peaks (photocurrents) P1, P2 may exceed the predetermined threshold value A1. This figure also shows the photocurrent (signal) B1 (photocurrent I3 at time t3) received by the computing device 180 from at least one of the one or more light receivers 120.The photocurrent (signal) B1 is generated by one or more fluorophores in the third sample 118c and may be proportional to the fluorescence detected by at least one of the one or more light receivers 120. B1 may not exceed a predetermined threshold value A1 or may not be equal to the threshold value. The computing device 180 may output information representing, for example, the comparison results of the signals (photocurrents) P1, P2 and / or B1.

[0121] FIG. 7 shows an embodiment of a method for detecting fluorescence of a sample obtained from a subject, the sample containing one or more fluorophores, representing a disease or a pathological condition, and the sample being carried out from the subject by a transparent conduit. The illustrated method may be implemented by an apparatus having a housing adapted to surround at least a part of the transparent conduit and a computing device. It is preferable to obtain from the subject a sample containing one or more fluorophores and representing a disease or a pathological condition. A sample containing one or more fluorophores is carried out from the subject by a transparent conduit at block 501. Specifically, this transportation may be achieved by negative pressure using a surgical suction instrument, an ultrasonic aspirator, or the like. The sample may be taken out from the subject using the same suction instrument, ultrasonic aspirator, or the like, or an instrument different from these. At block 510, light from one or more light sources may be emitted towards the sample carried out from the subject by the transparent conduit. At block 520, fluorescence generated by one or more fluorophores in the sample carried by the conduit may be detected by one or more light receivers. At block 530, a first signal corresponding to the photocurrent I (mA) and proportional to the detected fluorescence may be generated. The generated first signal may be received by computing means included in a computing device, and the computing device may be connected to a fluorescence assembly from at least one of the one or more light receivers. The first signal generated at block 530 may be compared with a predetermined threshold value regarding the first signal at block 540. At block 550, first information indicating the comparison result may be output. The output of the first information may be implemented by a user interface of the computing device, for example, by acoustic means, a display, an LED, tactile feedback, such as vibration, or a combination thereof.The first information may be output as one or more detected sounds including different volumes, timbres, and / or durations, or as one or more detected lights including different colors, luminances, and / or durations, or as a combination thereof, or as a combination of one or more detected sounds and one or more detected lights. Depending on the comparison result in block 540, if the first signal exceeds or is equal to a predetermined threshold, the first information output in block 550 may be different from the information output when the first signal does not exceed and is not equal to the predetermined threshold, according to the comparison result in block 540.

[0122] Additionally or alternatively, block 540 may include first and second expected thresholds for the first signal. Depending on the comparison result in block 540, if the first signal exceeds the first predetermined threshold and exceeds or is equal to the second predetermined threshold, the information output in block 550 may be different from the information output in block 550 when the first signal does not exceed or is not equal to at least one of the first and second predetermined thresholds, according to the comparison result in block 540.

[0123] Design changes to the embodiments of the invention described above can be made without departing from the scope of the invention as set forth in the appended claims. Expressions in the original specification used to describe and claim the invention, such as "including", "comprising", "incorporating", "have", "is", are to be construed as non-exclusive, i.e., items, components or elements not explicitly described may also be present. In the original specification, the singular form should also be construed as including the plural form.

Claims

1. An apparatus (100) for detecting fluorescence of a sample obtained surgically from a human or animal during fluorescence-guided surgery, substantially in real time, wherein the sample contains one or more fluorophores, the sample represents a disease or pathological condition, and during fluorescence-guided surgery, is a part or piece removed surgically from a human or animal, the part or piece including tissue, tumor, diffuse cancer type, and intracellular and extracellular body fluids, the sample being carried out of the human or animal by a transparent conduit (131) at a pressure lower than the ambient pressure during fluorescence-guided surgery, the apparatus comprising: - A housing (101) adapted to surround at least a portion of the transparent conduit (131), the housing (101) comprising: - One or more light sources (110) operable to emit light towards the sample carried in the transparent conduit (131); - One or more light receivers (120) operable to detect fluorescence produced by one or more fluorophores in the sample carried in the transparent conduit (131); - The one or more light sources (110) and the one or more light receivers (120) are arranged in horizontal, vertical or angled positions within the housing (101); - The housing (101) includes at least a first light source (110), the one or more light receivers (120) include at least a first light receiver (120) and a second light receiver (120), and the distance between the at least first light receiver (120) and the second light receiver (120) is selected from one or more distances based on the fluorescence induction rate, fluorescence decay, and fluorescence intensity of the sample containing one or more fluorophores, or a combination thereof; - The housing (101) has a computing device (180), the computing device (180) comprising: - Receiving a first signal from at least one of the one or more light receivers (120), the first signal being proportional to the fluorescence detected by the at least one of the one or more light receivers (120); - Comparing the first signal with a predetermined threshold value for the first signal; and - Computing means for outputting first information representing the result of the comparison, the apparatus providing substantially real-time detection of the fluorescence of the sample during fluorescence-guided surgery.

2. The apparatus according to claim 1, wherein the housing (101) is configured to surround the entire periphery of the transparent conduit (131).

3. The apparatus according to claim 1 or 2, wherein the housing (101) is formed of two parts attached to each other by attachment means.

4. The apparatus according to any one of claims 1 to 3, wherein the housing (101) is provided at longitudinal end portions of the housing (101) on opposite sides of each other and has a seal for bringing the housing into close contact with the transparent conduit (131).

5. The apparatus according to any one of claims 1 to 4, wherein the one or more light sources (110) can operate independently of each other to emit light of one or more fluorescence excitation curve wavelengths of the one or more fluorophores in the sample.

6. The apparatus according to any one of claims 1 to 5, wherein the one or more light receivers (120) can operate independently of each other to detect fluorescence of one or more wavelengths of one or more fluorescence emission curves of the one or more fluorophores in the sample.

7. The apparatus according to any one of claims 1 to 6, wherein each of the one or more light sources (110) further has one or more emission light filters (115) and / or each of the light receivers (120) has one or more incident light filters (125).

8. Each of the one or more fluorophores in the sample is independently selected from the group consisting of PpIX, indocyanine green, methylene blue, fluorescein, and salts, cyanines, T700, T800, BLZ-100, GB119, IRDye800CW conjugate, IRDye700Dx conjugate, EC17, LUM015, AVB-620, folic acid-fluorescein isothiocyanate (FITC), OTL38, gGlu-HMRG, green fluorophore conjugate, fluorescent dye-labeled peptide, fluorophore-conjugated antibody, fluorescent nanoparticles, activatable fluorescent probe, endogenous fluorophore, or combinations thereof. The apparatus according to any one of claims 1 to 7.

9. The apparatus according to any one of claims 1 to 8, wherein at least one of the one or more light sources is disposed in front of the one or more light receivers as viewed in the transport direction of the sample transported in the transparent conduit.

10. The housing (101) includes at least a first light source (110), and the one or more light receivers (120) include at least a first light receiver (120) and a second light receiver (120). The first light receiver (120) and the second light receiver (120) are positioned such that the distance from the first light receiver (120) to the first light source (110) is shorter than the distance from the second light receiver (120) to the first light source (110). The computing means further - receives a first signal from the first light receiver (120), the first signal being proportional to the fluorescence detected by the first light receiver (120), - determines a verification value for a second signal in response to receiving the first signal, - receives a second signal from the second light receiver (120), the second signal being proportional to the fluorescence detected by the second light receiver (120), - compares the second signal with the verification value, - outputs first information regarding the second signal in response to the second signal exceeding or being equal to the verification value, and - outputs second information regarding the second signal in response to the second signal not exceeding or not being equal to the verification value, thereby verifying the detection of fluorescence from the one or more light receivers (120) by the first light receiver (120). The device according to any one of claims 1 to 9.

11. The computing device (180) is provided within the housing (101). The device according to any one of claims 1 to 10.

12. A component kit comprising the device according to any one of claims 1 to 11 for detecting fluorescence of a sample obtained surgically from a human or animal during fluorescence-guided surgery, one or more fluorophores for use in combination with this device, and a transparent conduit (131).

Images