MULTIPURPOSE BIOPHOTON MEASURING DEVICE

Abstract

A multipurpose biophoton measuring device characterized in that it comprises: a. A UPE spectrograph. b. Two parallel chambers with charge-coupled devices (CCDs). c. Two large-aperture lenses. d. A multi-component cooling system for cooling the chambers to -100 °C. e. A computer for control, data acquisition, and processing, located in an adjacent room.

Description

MULTIPURPOSE BIOPHOTON MEASURING DEVICE TECHNICAL SECTOR The present invention pertains to the technical field of instrumental design of a biophoton emission (UPE) measuring device, primarily related to reactive oxygen species produced at the cellular level. The device combines the simultaneous acquisition of an image and the spectrum of the object being measured. More specifically, it combines spectroscopy and imaging, enabling the non-invasive analysis of fluids, tissues, microorganisms, animals, plants, and humans (focusing on a part or the entire body), which greatly expands its field of application in areas such as food, health, and defense and security, among others. BACKGROUND OF THE INVENTION Currently, the study of biophotonic emission is a largely unknown field in terms of applications because it needs technological developments and simple, robust, and versatile methodologies that allow its application and commercialization to companies / institutions in very diverse sectors. Considering a recent literature review analyzing the analytical aspects, results, and progress achieved in biophotonic emission studies in humans [1], it is noteworthy that two distinct instrumental designs have been employed to date. On the one hand, systems based on photomultiplier tubes (PMTs) have been used as a detection system, which has been the most widespread technology due to its high sensitivity in detecting minimal light emission. On the other hand, detection systems based on charge-coupled device (CCD) cameras have been used, which have been employed very sparingly for the two-dimensional visualization of spontaneous photoelectric emission (PEE) from different parts of the human body. With both detection systems, darkened rooms were used, as this is an extremely weak visible emission compared to external light.In fact, many authors used dark rooms painted black to avoid luminescence within the measurement area. One of the first instrumental designs developed, a 1954 prototype, consisted of an EMI 6260 PMT placed in a dark room to measure the biophotonic emission of various seeds (such as beans and lentils), but not of humans. The authors cooled the PMT to reduce the thermoelectric background and performed the experiments in darkness to avoid luminescence. With this setup, the authors were able to detect the light emitted by the seeds in the 450–600 nm range, and the signals were enhanced by adding a hydrogen peroxide solution to the pulps [2]. It wasn't until 1989 that Edwards and colleagues assembled one of the first prototypes for measuring spontaneous emission in humans. They used a By-Alkali 9814QB PMT in a darkened room with the ceiling, floor, and benches covered in matte black acrylic paint. Biophotonic emission was successfully measured from the abdomen, lower back, chest, forehead, and hands of two individuals. To avoid delayed luminescence (produced after excitation with external light), the subjects were protected from ambient light for one hour. The authors concluded that the intensity of biophotonic emission differed depending on the individual, obtaining large differences when their fronts were measured (23.47 ± 0.12 and 7.71 ± 0.15 counts per second (cps) for each individual). In addition, in this case they also used four bandpass filters to measure biophotonic emission at four wavelengths, observing that the emission was greater in the red region (600 nm) for both subjects [3]. Most subsequent designs have been based on Edward's prototype, considering the instrumental limitations of measuring biophotons due to their extremely weak signals, which require the use of external light protection procedures and measurements in darkened rooms. The prototype proposed by Kim et al. in 2002 showed an improvement by introducing a dark chamber within the darkened room to measure a finger with a PMT H6180-01. Despite requiring a darkened room, the use of smaller, easier-to-assemble dark chambers opens an opportunity for more practical biophoton measurements [4]. The instrumental design proposed by Yang et al. in 2015 consists of a dark chamber where the biophotonic emission of both hands was measured simultaneously with two PMT model 9235QA in a range of 290 to 630 nm. With this prototype, the authors measured the emission of two groups of individuals: healthy and diagnosed with a cold, finding significant differences with respect to the fingers whose signal was related in pairs [5]. However, in recent years, intensity measurements have been complemented by biophoton emission imaging of different parts of the human body. To this end, two different designs, from 2009 and 2016 respectively, used CCD cameras to measure the spontaneous biophoton emission of different parts of the human body. This technology was first used by Kobayashi et al. in the mid-2000s. The authors used a CCD camera (model SI 600) to record the spontaneous UPE emission of a subject throughout the day. By allowing CCD cameras to obtain two-dimensional information, the authors were able to display biophoton emission images of the individual throughout the day and were able to demonstrate once again the diurnal periodicity of this emission [6]. In 2016, Kobayashi et al. used the same CCD camera inside a dark chamber to measure, for the first time, the spectrum of a finger [7]. More recently, an electronically multiplied CCD (EMCCD) camera has been tested to measure the emission from a subject's hand [8]. This instrumental design was based on an iXon Ultra 888 EMCCD camera and a TEC-M55 mm tunable liquid crystal filter. Two different operating modes were tested: imaging and spectral distribution. Since the signal-to-noise ratio was very poor in imaging mode, full vertical clustering (FVB) mode was used to record the biophoton emission from a subject's hand, confirming the potential use of this technology to obtain spectral distributions based on the recorded wavelengths [8]. The potential of biophoton emission for measuring internal states has also been reviewed [9], observing that biophotons emitted increase in emotional states such as anger

[10] . Three prior inventions protecting Biophoton detector systems have also been located (CN203688443U, EP2728324A1 and EP0430150A2), but all differ in the technology employed from the present patent application. Thus, the utility model "Biological ultraweak photon radiation spectroscopic detector" (CN203688443U) is intended for the general measurement of ultraweak photon radiation (UPE) in biological samples. Regarding the instrument's construction, it differs in that it uses a light source (LED) as the radiation emitter, does not employ CCD cameras to obtain results, and uses a rotating optical filter plate and an LED driver module. In the present invention, the light source is the sample itself (the subjects of study), which is collected after a preliminary process that eliminates its delayed luminescence, which could interfere. The patent "Multi-spectral imaging method for ultraweak photon emission and system thereof" (EP2728324A1) describes a linear array of single-photon detectors for all wavelengths of response, combined with light-splitting technology. The two-dimensional multispectral ultraweak light imaging system achieves high-resolution optical modulation by adopting compression detection (CS) theory, digital light processing (DLP) technology, and using a linear array single-photon detector as the sensing element. The two-dimensional multispectral ultraweak light imaging system comprises a light filter, a first lens, a DMD control system, a second lens, a spectrophotometer, a linear array single-photon detector consisting of a plurality of detectable single photons with different wavelengths of response, and a central processing unit.The system's sensitivity can reach the level of a single photon. Finally, the patent "Method for testing quality and quality changes of biological systems and organochemical compositions interacting with these systems using measurements of ultraweak photon emission" (EP0430150A2) aims to measure the UPE in biological systems related to the food industry to determine their quality and changes in quality throughout the storage period. EXPLANATION OF THE INVENTION The present invention solves current problems regarding the need to improve the state of the art and its applications since, until now, versatile instrumental devices have not been developed in terms of the object of measurement and the measured signal (image and spectrum) nor have they been applicable to fields as diverse as food, health, defense and security, among other possibilities. This device is useful for measuring biophotons in fluids and biological samples, microorganisms, animals, plants or humans, considering it important that the design of the measurement module is versatile, simple, innocuous in appearance and convenient. This measuring device comprises the following components: a versatile measuring device located in a temperature-controlled room, within which there is a light-tight, darkened enclosure where the benchtop measuring system is placed. This system consists of two charge-coupled device (CCD) cameras with a cooling capacity down to -100 °C. One camera is connected to a spectrograph, which in turn is connected to a large-aperture lens

[11] to record spectra. Simultaneously, the other camera is connected directly to another large-aperture lens to record images at the same time as the first camera. The measurement room also contains elements necessary for the ultrasensitive detection of the device, such as a cooling system to ensure the optimal temperature of the CCD cameras. In an adjoining room is located the computer for controlling the device (the camera and spectrograph controller) and acquiring images and spectra. For the processing of image data, ad hoc algorithms have been developed that include the design of a mask (to extract only the region of interest from each image, ROI), the filtering and cleaning of the ROI, and the extraction of information from each image. For processing spectral data, ad hoc algorithms have been developed to extract useful information. These algorithms apply data preprocessing (baseline correction, normalization, smoothing, etc.) to the measured spectral profiles. In the case of human measurements, they also process other information (heart rate, dermal conductance, etc.) to simply and quickly provide a reliable discrimination, based on multivariate statistical analysis tools, of the spectral profiles and correlate the changes in the spectral profiles with other measured variables. In summary, the main advantages are as follows: - The use of CCD cameras allows for increased detection sensitivity since relatively long recording times are required (approximately between 10 and 30 min). - Unlike the vast majority of systems designed to date, the proposed device also makes it possible to simultaneously measure the image and biophotonic emission spectra. - The design of ad hoc algorithms allows processing the data obtained (image + spectrum) and obtaining relevant information from the selected image portion or the spectrum considered. - This device can be applied for the non-invasive analysis of fluids and biological samples, microorganisms, animals, plants and humans (a part or the whole body) facts that greatly expand its field of application. BRIEF DESCRIPTION OF THE FIGURES To complement the description being made and to help in a better understanding of the characteristics of the invention, the following figures are included, which, for illustrative and non-limiting purposes, represent the following: Figure 1: Measuring device consisting of the following parts: a UPE spectrograph (1.1); two CCD cameras (1.2 and 1.3); large aperture lenses (1.4 and 1.5); a computer for device control and image and spectral acquisition (1.6); and a cooling system (1.7). Figure 2: Experimental setup of the device in a dark room, with the following parts: a UPE spectrograph (1.1); two CCD cameras (1.2 and 1.3); and a large aperture lens (1.4). Figure 3: Schematic of the elements that make up the experimental triple refrigeration system mounted in the two circuits between the CCD chambers. The circuit consists of: two CCD chambers (1.2 and 1.3); a Peltier thermoelectric device (1.8) with a fan (below) and an extra water pump (above); a hermetically sealed water bath-pump with frozen polymers (1.9); and an open water bath with frozen polymers that can be quickly replaced (1.10). Figure 4: Elements that make up the experimental triple cooling system mounted in the two circuits between the CCD chambers. The cooling circuit itself (1.7) consists of: a Peltier thermoelectric device (1.8) with a fan at the top; a hermetically sealed water bath-pump with frozen polymers (1.9); and an open water bath with frozen polymers that can be quickly replaced (1.10). PREFERRED EMBODIMENT OF THE INVENTION A preferred embodiment of the present invention, according to Figures 1-4, comprises a multipurpose and highly versatile measuring device that allows measuring from people to animals, plants, microorganisms, or fluids and biological samples. The biophoton measuring device consists of a combination of elements that, when coupled, record the UPE (Ultra-Electron Potential) of the object being measured and allow simultaneous acquisition of its image and emission spectrum in the visible region. This requires: (i) a UPE spectrograph (1.1) that is completely isolated from outside light and that records the signal by means of a lens; (ii) two charge-coupled device (CCD) cameras (1.2 and 1.3) with cooling capacity down to -100 °C. One of them (1.2) is connected to the spectrograph (1.1), and the spectrograph is connected to a large-aperture lens for capturing spectra (1.4). In parallel, the other camera (1.3) is connected directly to another large-aperture lens (1.5) to capture images at the same time as the spectra are captured. (iii) A computer for device control and image and spectrum acquisition (1.6); (iv) and elements necessary for ultrasensitive detection of the device, such as the cooling system (1.7) to maintain a low temperature for the CCD cameras (Figures 3 and 4), the camera controller and its processor, and the spectrograph controller. The cooling system works by cooling water through two circuits connected in series with the cameras. One circuit recirculates the water through a Peltier thermoelectric device (1.8) and a sealed water bath with frozen polymers (1.9), and the other circuit uses a pump to recirculate the water through a heat exchanger immersed in an open water bath with quickly replaceable frozen polymers (1.10).The two independent water pumps ensure a continuous and adequate flow of water from the cooling systems to the CCD cameras and vice versa, carrying out repeated cycles to maintain the cameras at a temperature close to -100°C, where minimal thermal noise is produced. The entire device is isolated from light by means of a rigid, dark, wooden enclosure, capable of housing both the complete system in dark conditions and the object to be measured in front of the two lenses. The object to be measured is placed in an area that acts as a sampler for objects or even subjects. In the case of subjects, a chair / armchair has been chosen for the person's comfort during the measurement. A parabolic reflector can be placed behind the seat, which, depending on the subject, can be used to prevent the loss of biophotons, leading to the collection of a signal with a better signal-to-noise ratio. INDUSTRIAL APPLICATION This invention is susceptible to industrial application in very diverse fields such as health (measuring diseases in humans in a non-invasive way), food (differentiation between biological and non-biological foods or their state of preservation), laboratories (control and monitoring of microorganisms and biological fluids / samples), security and defense centers (monitoring false emotions during testimonies, dating of death by monitoring fluids or biological samples, etc.), among other possible applications. LITERATURE [1] Calcerrada, M.; García-Ruiz, C; Human Ultraweak Photon Emission: Key Analytical Aspects, Results and Future Progress, Critical Reviews in Analytical Chemistry, 2019;49, 368-381. [2] Colli, L.; Facchini, U.; Light emission by germinating plants, Il Nuovo Cimento (1943-1954). 1954, 12, 150-153. [3] Edwards, R.; Ibison, M. C.; Jessel-Kenyon, J.; Taylor, R. B. Measurements of human bioluminescence, Acupunct. Electrother. Res.1990, 15, 85-94. [4] Kim, T.; Nam, K.; Shin, H.; Lee, S.; Yang, J.; Soh, K. Biophoton emission from fingernails and fingerprints of living human subjects, Acupunct. Electrother. Res.2002, 27, 85-94. [5] Yang, M.; Pang, J.; Liu, J.; Liu, Y.; Fan, H.; Han, J. Spectral discrimination between healthy people and cold patients using spontaneous photon emission, Biom. Opt. Express. 2015, 6, 1331-1339. [6] Kobayashi, M.; Kikuchi, D.; Okamura, H. Imaging of Ultraweak Spontaneous Photon Emission from Human Body Displaying Diurnal Rhythm, Plos One.2009, 4, e6256. [7] Kobayashi, M.; Iwasa, T.; Tada, M. Polychromatic spectral pattern analysis of ultra- weak photon emissions from a human body, J. Photochem. Photobiol. B: Biology.2016, 159, 186-190. [8] Ortega-Ojeda, F.; Calcerrada, M.; Ferrero, A.; Campos, J.; Garcia-Ruiz, C. Measuring the Human Ultra-Weak Photon Emission Distribution Using an Electron- Multiplying, Charge-Coupled Device as a Sensor, Sensors.2018, 18, 1152. [9] Zapata Arráez, F.; Pastor Ruiz, V.; Ortega Ojeda, F.; Montalvo, G.; Ruiz Zolle, A.V.; García Ruiz, C. Human ultra-weak photon emission as non-invasive spectroscopic tool for diagnosis of internal states - A review. Journal of Photochemistr y and Photobiology B: Biology.2021, 216 / 112141.

[10] Zapata Arráez, F.; Pastor Ruiz, V.; Ortega Ojeda, F.; Montalvo, G; García Ruiz, C. Increment of spontaneous human biophoton emission caused by anger emotional states. Proof of concept. Microchemical Journal, 2021, 169, 106558.

[11] Smith, Warren Modern Lens Design 2005 McGraw-Hill.

Claims

1. A multipurpose biophoton measuring device characterized in that it comprises: a. A UPE spectrograph. b. Two parallel chambers with charge-coupled devices (CCDs). c. Two large-aperture lenses. d. A multi-component cooling system for cooling the chambers to -100 °C. e. A computer for control, data acquisition, and processing, located in an adjacent room.