The goal of this study is to explore the potential of pulsed and continuous wave light optogenetic stimulation on mouse embryonic cardiodynamics. Toward this goal, we engineered mouse embryos expressing the light-sensitive protein Channelrhodopsin-2 (ChR2) ubiquitously through the embryo. The embryos were dissected live and the optogenetic light stimulation of ChR2 at 473-nm was performed under imaging guidance. The pulsed stimulation allowed for a range of cardiodynamic behaviors and was overall found to have a milder effect on embryo viability, while the continuous wave stimulation provided an advantage in the faster mapping of optogenetic cardiac responses.
In the paper, the double-Debye and double-overdamped-oscillator models are considered for parametrization of the terahertz (THz) dielectric response of human brain tissues. Experimental data can be accurately reproduced for the intact tissues and gliomas using both models. While the double-Debye dielectric model is widely used in THz biophotonics, the double-overdamped-oscillator model appears to be more physically rigorous, since it satisfies the sum rule. In our opinion, the described double-overdamped-oscillator model is important for further research and development in THz biophotonics and neurodiagnosis of tumors.
In terahertz (THz) spectroscopy water content correlates with the tissue pathological changes. At the same time, water reduces the depth of THz-wave penetration in tissues. In order to unmask cells’ and tissues’ biophysical properties and to increase the tissues probing depth, immersion optical clearing (IOC) was recently introduced in the THz range. For studying common IOC agent’s optical properties in the frequency range of 0.1–2.5 THz, pulsed spectroscopy was used. Diffusion coefficients of IOC agents in ex vivo rat brain tissue were studied using the collimated transmission spectroscopy in the visible range. Two-dimensional nomogram was used to objectively compare IOC agents, based on their THz-wave absorption coefficients and diffusion rates.
Terahertz (THz) spectroscopy and imaging allow to differentiate between healthy tissues and tumors of different types, due to keen sensitivity to the water content. However, a high absorption of THz waves by water molecules limits application of THz technology for medical diagnosis. Among the existing methods for tissue clearing only immersive optical clearing (IOC) is acceptable for in vivo study. In order to choose optimal agents for IOC, we reconstructed THz dielectric properties of common agents using methods of THz pulsed spectroscopy and we measured diffusion coefficients of agents in rat brain analyzing collimated transmittance in visible range.
Application of a fiber Fabry–Perot interferometer for studying the sound response of cellular structures and aqueous solutions is discussed. Aqueous suspensions of baker’s yeast and commercial natural drinking water, treated by electrolysis, were applied as models in order to mimic processes in biological tissues and liquids. The distribution of frequency intensities in the acoustic spectrograms yields evaluation of a biological system response to the external exposure. In a yeast suspension, few minutes after the sound irradiation with the frequency of 3 kHz and the sound pressure of 50 to 60 dB, we observed regular fluctuations in the output acoustic signal, with the maximal period of about 100 s. Furthermore, a sound response in the frequency range of 400 to 600 Hz maintained in signals for few minutes after the exposure. The observed results demonstrate that the proposed interferometric sensor has strong potential in biology and medicine since it is quite simple, portable, and highly sensitive device for analyses the sound response of a living system.
We applied reflection-mode terahertz (THz) pulsed spectroscopy to study ex vivo the optical properties of human brain tumors with the different World Health Organization grades, as well as of perifocal regions comprised of intact (healthy) and edematous tissues. We applied gelatin-embedding in order to fix freshly-excised tissues, thus, preserving them from hydration/dehydration and sustaining their THz response unaltered for a couple of hours after resection. We observed a contrast between the THz optical properties of intact tissues and tumors, including gliomas and meningiomas of the brain, in turn, the response of edematous tissues is close to that of a tumor. The observed contrast between intact tissues and tumors has an endogenous character and originates reportedly from increased water content in a tumor due to edema, abnormal vascularity and, in some cases, necrotic debris. The observed results justify a prospect of THz technology in the intraoperative label-free diagnosis of human brain tumors.
An intraoperative diagnosis of brain tumors is one of the most urgent and challenging problem of the modern neurosurgery. The most important measure of the effectiveness of treatment is the complete tumor resection. The existing methods of the intraoperative neurodiagnosis of tumors are plagued with limited sensitivity, especially for low-grade gliomas, and, furthermore, can remain rather expensive. The recently reported results of optical coherence tomography (OCT) application for finding differences between healthy and malignant tissues make it become one of the promising label-free diagnostic instruments. Nevertheless, the wide use of OCT in clinical practice is limited by the lack of complete study of its opportunities in neurosurgery, which leads to a huge scientific interest. Our research, aimed at the study of the ability of OCT for the intraoperative diagnosis of brain gliomas of different grades, has the goal to observe the differences between OCT signals obtained for ex vivo samples of various types of human brain glioma and intact brain tissue. We propose a 3D-feature based data analysis that demonstrates promising results in differentiation of tissue classes.
We developed a method for reconstructing the THz dielectric response of a thin liquid sample. A self-made sample cuvette was designed for the transmission-mode THz pulsed spectroscopy of liquids. Numerical simulations and theoretical studies of the proposed reconstruction procedure were performed in order to optimize the sample geometry and predict uncertainties in reconstructed dielectrical properties. A number of agents for immersion optical clearing of tissues was studied using the proposed method in the THz range. The developed method can be applied for all types of sufficiently transparent liquid samples.
We performed the transmission-mode terahertz (THz) pulsed spectroscopy of several THz-wave penetration- enhancing agents (PEAs): glycerol, propylene glycol, ethylene glycol, and polyethylene glycol, featuring the molecular weight of 200, 300 and 400. We vacuumized the THz beam path in order to reduce an impact of water vapor on measured data. We reconstructed optical properties and dielectric constants of the abovementioned PEAs in the spectral range of 0.1 to 2.5 THz. We analyzed measured THz optical properties along with the literature data for coefficients of PEAs' diffusion into tissues in order to objectively uncover strength and weaknesses of their use in the immersion optical clearing of tissues at THz frequencies.
Intraoperative diagnosis of brain tumors remains a challenging problem of modern neurosurgery. A complete resection of tumor is the most important factor, determining an efficiency of its treatment, while an incomplete resection, caused by inaccurate detection of tumor margins, increases a probability of the tumor recurrence. The existing methods of the intraoperative neurodiagnosis of tumors are plagued with limited sensitivity and specificity; they remain laborious, time-consuming and/or rather expensive. Therefore, the development of novel methods for the intraoperative diagnosis of gliomas relying on modern instruments of medical imaging is a topical problem of medicine, physics, and engineering. In our research, we studied the ability of dual-modality imaging that combines such methods as optical coherence tomography (OCT) and terahertz (THz) pulsed spectroscopy, for intraoperative diagnosis of brain tumors with a strong emphasize on a human brain gliomas. We performed experimental studies of the frequency-dependent THz dielectric properties and OCT imaging of healthy (intact) and pathological brain tissues ex vivo in order to analyze the prospect for differentiation between tissue classes. The observed results highlight a potential of the considered instruments in the label-free intraoperative neurodiagnostics.
We applied terahertz (THz)-pulsed spectroscopy to study ex vivo the refractive index and absorption coefficient of human brain gliomas featuring different grades, as well as perifocal regions containing both intact and edematous tissues. Glioma samples from 26 patients were considered and analyzed according to further histological examination. In order to fix tissues for the THz measurements, we applied gelatin embedding, which allows for sustaining their THz response unaltered, as compared to that of the freshly excised tissues. We observed a statistical difference between the THz optical constants of intact tissues and gliomas of grades I to IV, while the response of edema was similar to that of tumor. The results of this paper justify a potential of THz technology in the intraoperative label-free diagnosis of human brain gliomas for ensuring the gross-total resection.
Application of terahertz (THz) spectroscopy for biological tissues is strongly limited by the extremely low penetration depth due to THz absorption by tissue water. One of the possible solution of such problem is the usage of THz wave penetration-enhancing agents (PEA) for optical clearing of tissues. In the present paper, the transmission-mode THz spectroscopy of a set of PEAs (polyethylene glycol with different molecular weight, propylene glycol, ethylene glycol, and dimethyl sulfoxide) was performed in order to reconstruct their dielectric properties and compare them with that of water. The obtained results emphasize the feasibility of using PEG to enhance the depth of THz wave penetration into tissues.
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