植物幼苗原发性超弱光子成像的研究

采用光子计数成像系统（PIAS）对植物幼苗萌发过程的超弱发光进行观察。结果表明,自发光子长时间积累可形成二维图象;光子计数和采集图象均可得到植物体的自发发光;通过实验探测到幼苗的根,叶在同一平面图象有不同的发光表现;光子成像系统可客观地比较生物自发超弱发光,为进一步研究超弱发光机理提供实验基础。     

Chemiluminescence of cereal products. II. Chemiluminescence spectra

Spectra of ultraweak chemiluminescence (CL) accompanying auto-oxidation and hydration of cereal products have been measured using single photon counting and cut-off filters. The spectra cover the 380–880 nm spectral range with maxima centred around 600 nm. Analytically pure air-dried carbohydrates like agar, cellulose and nitrocellulose give emission too weak for spectral measurements. The emission from water pure carbohydrates is on average 4–12 times higher and emission spectra are similar to those from cereal products. The effect of free radical scavengers, SOD and O*₂ (¹Δ_g)-quenchers on CL spectra indicates a contribution of radical reactions with the participation of excited carbonyls, O₂⁻ and excited molecular oxygen dimoles. Moreover, possible mechanisms of chemi-excitation due to a cooperative H-bond formation during the hydration of carbohydrates and/or recombination of trapped radicals and electron-holes are discussed. It is also postulated that the excitation energy transfer to natural sensitizers occuring in cereal products may account for non-specific broad spectra and differences in the intensity of CL. © 1998 John Wiley & Sons, Ltd.     

Imaging of Ultraweak Spontaneous Photon Emission from Human Body Displaying Diurnal Rhythm

The human body literally glimmers. The intensity of the light emitted by the body is 1000 times lower than the sensitivity of our naked eyes. Ultraweak photon emission is known as the energy released as light through the changes in energy metabolism. We successfully imaged the diurnal change of this ultraweak photon emission with an improved highly sensitive imaging system using cryogenic charge-coupled device (CCD) camera. We found that the human body directly and rhythmically emits light. The diurnal changes in photon emission might be linked to changes in energy metabolism.     

Light emission from tryptophan oxidation by hypobromous acid

The emission of ultraweak light from cells is a phenomenon associated with the oxidation of biomolecules by reactive oxygen species. The indole moiety present in tryptophan, serotonin and melatonin is frequently associated with the emission of light during the oxidation of these metabolites. This study presents results for hypobromous acid (HOBr) oxidation of tryptophan as a putative endogenous source of ultraweak light emission. We found that chemiluminescence elicited by the oxidation of tryptophan by HOBr was significantly higher than by hypochlorous acid (HOCl). This difference was related to secondary oxidation reactions, which were more intense using HOBr. The products identified during oxidation by HOCl, but depleted by using HOBr, were N‐formylkynurenine, kynurenine, 1,2,3,3a,8,8a‐hexahydro‐3a‐hydroxypyrrolo[2,3‐b]‐indole‐2‐carboxylic acid, oxindolylalanine and dioxindolylalanine. The emission of light is dependent on the free α‐amino group of tryptophan, and hence, the indole of serotonin and melatonin, although efficiently oxidized, did not produce chemiluminescence. The emission of light was even greater using taurine monobromamine and dibromamine as the oxidant compared to HOBr. A mechanism based on bromine radical intermediates is suggested for the higher efficiency in light emission. Altogether, the experimental evidence described in the present study indicates that the oxidation of free tryptophan or tryptophan residues in proteins is an important source of ultraweak cellular emission of light. This light emission is increased in the presence of taurine, an amino acid present in large amounts in leukocytes, where this putative source of ultraweak light emission is even more relevant. Copyright © 2012 John Wiley & Sons, Ltd.     

Biophoton emission of human body

For the first time systematic measurements of the "ultraweak" photon emission of the human body (biophotons) have been performed by means of a photon detector device set up in darkness. About 200 persons have been investigated. In a particular case one person has been examined daily over several months. It turned out that this biophoton emission reflects, (i) the left-right symmetry of the human body; (ii) biological rhythms such as 14 days, 1 month, 3 months and 9 months; (iii) disease in terms of broken symmetry between left and right side; and (iv) light channels in the body, which regulate energy and information transfer between different parts. The results show that besides a deeper understanding of health, disease and body field, this method provides a new powerful tool of non-invasive medical diagnosis in terms of basic regulatory functions of the body.     

淡水鱼超微弱光子辐射的实验研究

采用一台高灵敏的生物活体单光计数系统对淡水鱼的主要超微弱发光特性进行了研究.淡水鱼的各种不同脏器的光子辐射强度数值为3.6—97.3CPS.发射光谱的最大值接近640nm处和460nm附近.实验结果表明,超微弱光子辐射的总强度随所测的时间而变化,加入UOUO_2~( )离子之后,其发光强度下降.     

Delayed Fluorescence and Ultraweak Light Emission from Isolated Chloroplasts (Comparison of Emission Spectra and Concentration Dependence)

The ultraweak light emission of isolated chloroplasts (Hidegand Inaba (1991) Photochem. Photobiol. 52: 137) was investigatedin comparison to delayed light emission. We compared the concentrationdependence and the spectral distribution of the light emittedfrom isolated chloroplasts stored in the dark for 10 s, 2 min(delayed light emission), 4 and 10 h (ultraweak light emission),respectively. In samples with low chlorophyll concentration, spectra of allemission phenomena were maximal at 685–695 nm, but spectraof ultraweak light, especially that of long term (10 h) emission,were broader in the 700–800 nm region than spectra ofdelayed light, indicating emission from a bigger variety ofchlorophyll molecules. The intensity of delayed light and short term (4 h) ultraweaklight exhibited a simple, saturating exponential dependenceon chlorophyll concentration, while long term (10 h) ultraweaklight emission was best described as a saturating exponentialcontaining a quadratic function of the concentration. This differencesuggests that long term ultraweak light emission is broughtabout by reactions distinct from the earlier described mechanismof electron transport related dark photoemission. (Received November 15, 1991; Accepted May 18, 1992)     

Lightsheet fluorescence lifetime imaging microscopy with wide‐field time‐correlated single photon counting

We report on wide‐field time‐correlated single photon counting (TCSPC)‐based fluorescence lifetime imaging microscopy (FLIM) with lightsheet illumination. A pulsed diode laser is used for excitation, and a crossed delay line anode image intensifier, effectively a single‐photon sensitive camera, is used to record the position and arrival time of the photons with picosecond time resolution, combining low illumination intensity of microwatts with wide‐field data collection. We pair this detector with the lightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradients in human cancer cell spheroids, and C. elegans.     

Superweak irradiation of marine invertebrates

The inhibitory analysis of the spontaneous ("ultraweak") and the luminole-induced chemiluminescence of marine Sycon sponges and Aiptasia actinias supports the idea that ultraweak photon emission of marine invertebrates is a consequence of Ca(2+)-dependent processes related to the interaction of reactive oxygen species with some endogenous fluorophore substrates.     

Activity-dependent neural tissue oxidation emits intrinsic ultraweak photons

Living organisms have been known to spontaneously emit ultraweak photons in vivo and in vitro. Origin of the photon emission remains unclear, especially in the nervous system. The spontaneous ultraweak photon emission was detected here from cultured rat cerebellar granule neurons using a photomultiplier tube which was highly sensitive to visible light. The photon emission was facilitated by the membrane depolarization of neurons by a high concentration of K+ and was attenuated by application of tetrodotoxin or removal of extracellular Ca2+, indicating the photon emission depending on the neuronal activity and likely on the cellular metabolism. Furthermore, almost all the photon emission was arrested by 2,4-dinitrophenylhydrazine, indicating that the photon emission would be derived from oxidized molecules. Detection of the spontaneous ultraweak photon emission will realize noninvasive and real-time monitoring of the redox state of neural tissue corresponding to the neuronal activity and metabolism.     

Imaging of ultra‐weak bio‐chemiluminescence and singlet oxygen generation in germinating soybean in response to wounding

Ultra‐weak bio‐chemiluminescence (UBC) from germinating soybean (Glycine max L. Merr) cotyledon under mechanical wounding was observed using a high‐sensitivity imaging system based on an ICCD detector and a highly sensitive single photon counter (SPC) device. The UBC imaging showed that the intensity at the injury location on a wounded cotyledon was obviously enhanced as compared with that at the non‐injured point. The UBC intensity of wounded cotyledons was initially very high and reached a stationary state after about 5 min. Wounding‐induced emission could be suppressed by wounding in the presence of sodium azide. Deuterium oxide amplified the emission intensity. It was concluded that singlet oxygen (¹O₂) was the main cause of the emission during the wounding phase. Copyright © 2002 John Wiley & Sons, Ltd.     

Highly sensitive determination of transient generation of biophotons during hypersensitive response to cucumber mosaic virus in cowpea

The hypersensitive response (HR) is one mechanism of the resistance of plants to pathogen infection. It involves the generation of reactive oxygen species (ROS) which have crucial roles in signal transduction or as toxic agents leading to cell death. Often, ROS generation is accompanied by an ultraweak photon emission resulting from radical reactions that are initiated by ROS through the oxidation of living materials such as lipids, proteins, and DNA. This photon emission, referred to as 'biophotons', is extremely weak, but, based on the technique of photon counting imaging, a system has been developed to analyse the spatiotemporal properties of photon emission. Using this system, the dynamics of photon emission which might be associated with the oxidative burst, which promotes the HR, have been determined. Here, the transient generation of biophotons is demonstrated during the HR process in cowpea elicited by cucumber mosaic virus. The distinctive dynamics in spatiotemporal properties of biophoton emission during the HR expression on macroscopic and microscopic levels are also described. This study reveals the involvement of ROS generation in biophoton emission in the process of HR through the determination of the inhibitory effect of an antioxidant (Tiron) on biophoton emission.     

Biophoton emission

The phenomenon of ultraweak photon emission from living systems was further investigated in order to elucidate the physical properties of this radiation and its possible source. We obtained evidence that the light has a high degree of coherence because of (1) its photon count statistics, (2) its spectral distribution, (3) its decay behavior after exposure to light illumination, and (4) its transparency through optically thick materials. Moroever, DNA is apparently at least an important source, since conformational changes induced with ethidium bromide in vivo are clearly reflected by changes of the photon emission of cells. The physical properties of the radiation are described, taking DNA as an exciplex laser system, where a stable state can be reached far from thermal equilibrium at threshold.     

Correlation between hydroperoxide-induced chemiluminescence of the heart and its function

The isolated perfused rat heart emits a spontaneous ultraweak chemiluminescence. When the perfusion is stopped, light emission decreases, indicating the dependency of this phenomenon on aerobic metabolism. Emitted chemiluminescence was markedly enhanced following perfusion with 0.05 mM H₂O₂ or cumene hydroperoxide or tert-butyl hydroperoxide; substitution of O₂ for N₂ in the gassing mixture of the perfusion media significantly lowered photon emission. Lipid peroxidation, which is known to be associated with chemiluminescence, was evaluated by HPLC analysis of peroxidized and unperoxidized heart phosphatidylcholines. During hydroperoxide perfusion, coronary flow and heart rate progressively decreased, while lactic dehydrogenase was released after complete cardiac arrest. The resultant morphology of this damage corresponds to the so-called ‘stone heart’, a pattern already described in both human and experimental pathology.     

Ultraweak photoemission from dark-adapted leaves and isolated chloroplasts

Dark-adapted isolated spinach chloroplasts and leaves, unlike sub-chloroplast fractions, are capable of emitting ultraweak light spontaneously (50-125 counts/s per cm2). The emission of leaves is due to two processes with activation energies of 97 and 25 kJ/mol while in isolated chloroplasts, it is the result of a single process (98 kJ/mol), as indicated by the Arrhenius plots of the intensity. Emission spectra demonstrate that the terminal step of these reactions is the excitation of chlorophyll in both samples. We suggest that the additional component in the ultraweak light emission of leaves may be related to mitochondria.     

Biophoton emission. New evidence for coherence and DNA as source

The phenomenon of ultraweak photon emission from living systems was further investigated in order to elucidate the physical properties of this radiation and its possible source. We obtained evidence that the light has a high degree of coherence because of (1) its photon count statistics, (2) its spectral distribution, (3) its decay behavior after exposure to light illumination, and (4) its transparency through optically thick materials. Moreover, DNA is apparently at least an important source, since conformational changes induced with ethidium bromide in vivo are clearly reflected by changes of the photon emission of cells. The physical properties of the radiation are described, taking DNA as an exciplex laser system, where a stable state can be reached far from thermal equilibrium at threshold.     

A photon counting device for the measurement of nanosecond and microsecond kinetics of light emission from biological systems

This paper describes the instrumentation and operation of a photon counting system which was constructed to measure the delayed light emission from the algae Scenedesmus. The functioning of the system was illustrated by a plot which compared data of the recorded delayed light emission with data of the system noise over the first 20-μsec interval after extinction of a modulated laser beam. We believe that this paper shows the utility of photon counting as a satisfactory method of measuring biological light emission, especially a very rapid process such as the delayed light emission from photosynthetic organisms.     

水稻籽粒灌浆过程中超弱发光特性的研究

本实验对水稻籽粒灌浆过程中不同时期籽粒进行了超弱发光测定。实验结果表明：在整个灌浆过程中单粒发光量呈单峰曲线,在籽粒全部充实并且颜色由青转黄时,发光量最大;发光强度呈递减趋势;运用该技术有可能进行水稻不同品种间的比较和品种灌浆力强弱的筛选。     

Comparison of seven bio- and chemiluminescent reagents for in situ detection of antigens and nucleic acids

Bio- and chemiluminescence have proved sensitive enough to compete with chromogenic and radioisotopic tracers for in situ detection. However, they must also provide a discriminant morphological analysis of the specific signal. We have tested seven bio-or chemiluminescent reagents for tissue antigen and nucleic acid detection by immunocytochemistry (ICC) or in situ hybridization (ISH). They were based on luminescent detection of peroxidase, aikaline phosphatase, β-galactosidase or xanthine oxidase. We also explored whether high molecular weight polymers could increase the spatial definition of the photon emission. An ICCD camera was used to collect the light signal provided by immunolabelling of endothelial cells and by ISH of human papilloma virus on cell smears. Among the enzyme-luminescent substrate combinations tested, the enhanced luminol chemiluminescence (ECL) gave the best resolution of the specific signal. The other systems were mainly hampered by a high diffusion of the reaction product over the tissue section. Unfortunately, in this case, the high molecular weight polymers tested were inefficient. However, the addition of polyvinylalcohol (PVA) or polyvinylpyrrolidone (PVP) significantly improved respectively the definition and intensity of ECL photon emission. We demonstrate that chemiluminescence gives a morphological resolution allowing histological examination. The extension of this new application, now depends on physicochemical adaptation of chemiluminescent reagents to the constraints of tissue detection.     

Light from maillard reaction: Photon counting,emission spectrum,photography and visual perception

Several authors have reported on high-sensitivity measurement of oxygen-dependent low-level chemiluminescence (CL) from Maillard reactions (MR), i.e. nonenzymatic amino-carbonyl reactions between reducing sugars and amino acids (also referred to as nonenzymatic browning). Here we report for the first time, that light from Maillard reactions can be seen by the human eye and also can be photographed. In parallel with visual perception and photography CL was monitored by means of a CL-detection programme of a liquid scintillation counter (LSC, single photon rate counting). CL emission spectrum was recorded by a monochromator-microchannel plate photomultiplier arrangement. CL intensity from reaction of 6-aminocaproic acid with D-ribose (200 mg each) in 5 mL H₂O at pH 11 at 95°C was high enough for visual perception after adaptation to absolute darkness. Reaction in dimethylsulphoxide (DMSO) exhibited strongly enhanced CL (10 mg each in 5 mL were sufficient for visual detection) and could be photographed (15 minutes' exposure, ASA 6400); all characteristics of Maillard specific CL (O₂-dependence, no CL from nonreducing sugars, inhibition by sulphur compounds) remained. Visual detection of CL and measurement by LSC were in full concordance. The CL emission spectrum showed two broad peaks at around 500 nm and 695 nm. Fluorescence emission of the brown reaction mixture matched the bluegreen part of the CL emission spectrum. Emission of visible light during Maillard reactions may partly originate from oxygen-dependent generation of excited states and energy transfer to simultaneously formed fluorescent products of the browning reaction.