Transient photothermal inactivation of Escherichia coli stained with visible dyes by using a nanosecond pulsed laser

Kohmura, Yuji; Igami, Natsuho; Tatsuno, Ichiro; Hasegawa, Tadao; Matsumoto, Takahiro

doi:10.1038/s41598-020-74714-5

Cited by 7 publications

(11 citation statements)

References 51 publications

(69 reference statements)

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“…Here, GNPs of various diameters were photothermally melted by instantaneous pulse laser irradiation, and the transient photothermal behaviour of GNP colloids versus the diameter was elucidated based on both the optical absorption spectrum and electron microscopy images. We consider that the results of our previous study 40 , in which the transient temperature behaviour had been theoretically estimated, are now experimentally confirmed in this work.…”

Section: Introductionsupporting

confidence: 80%

“…We have previously evaluated the maximum temperatures of GNPs as a function of diameter and absorbed laser fluence (mJ/cm 2 ) 40 . The transient response of the temperature (local temperature) as a function of time, t , can be given as: …”

Section: Resultsmentioning

confidence: 99%

“…To overcome the drawbacks of the fs laser inactivation methods described above, we have demonstrated the efficient inactivation of dye-infused, micrometre-sized bacteria by using a low-power and easily available nanosecond (ns) visible pulse laser (532 nm) and obtained 3-log inactivation of Escherichia coli ( E. coli ) bacteria over a short period of treatment time, i.e., within 20 min 40 . In that study, the inactivation mechanism was analysed based on a simple optical absorption effect, which induces transient photothermal evaporation in E. coli bacteria.…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we found that the scattering effect 41 , 42 , which was not previously discussed, plays an important role in inactivation because the input irradiation is inherently scattered by the bacteria; therefore, the dose that bacteria feel is reduced by scattering. To investigate both the scattering and absorption effects for various wavelength regions, we fused visible light absorbing dye with E. coli 40 , because the refractive index as well as the extinction coefficient can be modified by this staining treatment. The complex inactivation behaviours can be clearly shown by comparative experiments between unstained and stained E. coli with site-selective (various wavelength) irradiation obtained by a wavelength tunable pulsed laser.…”

Section: Introductionmentioning

confidence: 99%

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Mechanism of transient photothermal inactivation of bacteria using a wavelength-tunable nanosecond pulsed laser

Tatsuno

Niimi

Tomita

et al. 2021

Sci Rep

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There is a great demand for novel disinfection technologies to inactivate various pathogenic viruses and bacteria. In this situation, ultraviolet (UVC) disinfection technologies seem to be promising because biocontaminated air and surfaces are the major media for disease transmission. However, UVC is strongly absorbed by human cells and protein components; therefore, there are concerns about damaging plasma components and causing dermatitis and skin cancer. To avoid these concerns, in this study, we demonstrate that the efficient inactivation of bacteria is achieved by visible pulsed light irradiation. The principle of inactivation is based on transient photothermal heating. First, we provide experimental confirmation that extremely high temperatures above 1000 K can be achieved by pulsed laser irradiation. Evidence of this high temperature is directly confirmed by melting gold nanoparticles (GNPs). Inorganic GNPs are used because of their well-established thermophysical properties. Second, we show inactivation behaviour by pulsed laser irradiation. This inactivation behaviour cannot be explained by a simple optical absorption effect. We experimentally and theoretically clarify this inactivation mechanism based on both optical absorption and scattering effects. We find that scattering and absorption play an important role in inactivation because the input irradiation is inherently scattered by the bacteria; therefore, the dose that bacteria feel is reduced. This scattering effect can be clearly shown by a technique that combines stained Escherichia coli and site selective irradiation obtained by a wavelength tunable pulsed laser. By measuring Live/Dead fluorescence microscopy images, we show that the inactivation attained by the transient photothermal heating is possible to instantaneously and selectively kill microorganisms such as Escherichia coli bacteria. Thus, this method is promising for the site selective inactivation of various pathogenic viruses and bacteria in a safe and simple manner.

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Section: Introductionsupporting

confidence: 80%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mechanism of transient photothermal inactivation of bacteria using a wavelength-tunable nanosecond pulsed laser

Tatsuno

Niimi

Tomita

et al. 2021

Sci Rep

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“…First, to confirm the characteristics of the effects of the 222-UVC radiation used in this study on biological organisms, the effects on the viability of E. coli and P1 phages, which have been widely reported thus far [ 16 – 19 ], were investigated. E. coli and P1 phages were subjected to a different dose (0–150 J m −2 ) of 254-UVC or 222-UVC radiation, and the survival rate of E. coli (Fig.…”

Section: Resultsmentioning

confidence: 99%

222 nm ultraviolet radiation C causes more severe damage to guard cells and epidermal cells of Arabidopsis plants than does 254 nm ultraviolet radiation

Otake

Yoshiyama

Yamaguchi

et al. 2021

Photochem Photobiol Sci

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Lamps that emit 222 nm short-wavelength ultraviolet (UV) radiation can be safely used for sterilization without harming human health. However, there are few studies on the effects of 222 nm UVC (222-UVC) radiation exposure on plants compared with the effects of germicidal lamps emitting primarily 254 nm UVC (254-UVC) radiation. We investigated the growth inhibition and cell damage caused by 222-UVC exposure to Arabidopsis plants, especially mitochondrial dynamics, which is an index of damage caused by UVB radiation. Growth inhibition resulted from 254-UVC or 222-UVC exposure depending on the dose of UVC radiation. However, with respect to the phenotype of 222-UVC-irradiated plants, the leaves curled under 1 kJ m−2 and were markedly bleached under 10 kJ m−2 compared with those of plants irradiated with 254-UVC. The cellular state, especially the mitochondrial dynamics, of epidermal and mesophyll cells of Arabidopsis leaves exposed to 254-UVC or 222-UVC radiation was investigated using Arabidopsis plants expressing mitochondrial matrix-targeted yellow fluorescent protein (MT-YFP) under the control of Pro35S to visualize the mitochondria. 222-UVC (1 or 5 kJ m−2) severely damaged the guard cells within the epidermis, and YFP signals and chloroplast autofluorescence in guard cells within the epidermis exposed to 222-UVC (1 or 5 kJ m−2) were not detected compared with those in cells exposed to 254-UVC radiation. In addition, 222-UVC irradiation led to mitochondrial fragmentation in mesophyll cells, similar to the effects of 254-UVC exposure. These results suggest that 222-UVC severely damages guard cells and epidermal cells and that such damage might have resulted in growth inhibition.

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Time-dose reciprocity mechanism for the inactivation of Escherichia coli explained by a stochastic process with two inactivation effects

Matsumoto

Tatsuno

Yoshida

et al. 2022

Sci Rep

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There is a great demand for developing and demonstrating novel disinfection technologies for protection against various pathogenic viruses and bacteria. In this context, ultraviolet (UV) irradiation offers an effective and convenient method for the inactivation of pathogenic microorganisms. The quantitative evaluation of the efficacy of UV sterilization relies on the simple time-dose reciprocity law proposed by Bunsen-Roscoe. However, the inactivation rate constants reported in the literature vary widely, even at the same dose and wavelength of irradiation. Thus, it is likely that the physical mechanism of UV inactivation cannot be described by the simple time-dose reciprocity law but requires a secondary inactivation process, which must be identified to clarify the scientific basis. In this paper, we conducted a UV inactivation experiment with Escherichia coli at the same dose but with different irradiances and irradiation durations, varying the irradiance by two to three orders of magnitude. We showed that the efficacy of inactivation obtained by UV-light emitting diode irradiation differs significantly by one order of magnitude at the same dose but different irradiances at a fixed wavelength. To explain this, we constructed a stochastic model introducing a second inactivation rate, such as that due to reactive oxygen species (ROS) that contribute to DNA and/or protein damage, together with the fluence-based UV inactivation rate. By solving the differential equations based on this model, the efficacy of inactivation as a function of the irradiance and irradiation duration under the same UV dose conditions was clearly elucidated. The proposed model clearly shows that at least two inactivation rates are involved in UV inactivation, where the generally used UV inactivation rate does not depend on the irradiance, but the inactivation rate due to ROS does depend on the irradiance. We conclude that the UV inactivation results obtained to date were simply fitted by one inactivation rate that superimposed these two inactivation rates. The effectiveness of long-term UV irradiation at a low irradiance but the same dose provides useful information for future disinfection technologies such as the disinfection of large spaces, for example, hospital rooms using UV light, because it can reduce the radiation dose and its risk to the human body.

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Transient photothermal inactivation of Escherichia coli stained with visible dyes by using a nanosecond pulsed laser

Cited by 7 publications

References 51 publications

Mechanism of transient photothermal inactivation of bacteria using a wavelength-tunable nanosecond pulsed laser

Mechanism of transient photothermal inactivation of bacteria using a wavelength-tunable nanosecond pulsed laser

222 nm ultraviolet radiation C causes more severe damage to guard cells and epidermal cells of Arabidopsis plants than does 254 nm ultraviolet radiation

Time-dose reciprocity mechanism for the inactivation of Escherichia coli explained by a stochastic process with two inactivation effects

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