Exosomal membrane proteins analysis using a silicon nanowire field effect transistor biosensor.

Exosomes are of great significance in clinical diagnosis, due to their high homology with parental generation, which can reflect the pathophysiological status. However, the quantitative and classification detection of exosomes is still faced with the challenges of low sensitivity and complex operation. In this study, we develop an electrical and label-free method to directly detect exosomes with high sensitivity based on a Silicon nanowire field effect transistor biosensor (Si-NW Bio-FET). First, the impact of Debye length on Si-NW Bio-FET detection was investigated through simulation. The simulation results demonstrated that as the Debye length increased, the electrical response to Si-NW produced by charged particle at a certain distance from the surface of Si-NW was greater. A Si-NW Bio-FET modified with specific antibody CD81 on the nanowire was fabricated then used for detection of cell line-derived exosomes, which achieved a low limit of detection (LOD) of 1078 particles/mL in 0.01 × PBS. Furthermore, the Si-NW Bio-FETs modified with specific antibody CD9, CD81 and CD63 respectively, were employed to distinguish exosomes derived from human promyelocytic leukemia (HL-60) cell line in three different states (control group, lipopolysaccharide (LPS) inflammation group, and LPS + Romidepsin (FK228) drug treatment group), which was consistent with nano-flow cytometry. This study provides a highly sensitive method of directly quantifying exosomes without labeling, indicating its potential as a tool for disease surveillance and medication instruction.

Volatile carbonyl metabolites analysis of nanoparticle exposed lung cells in an organ-on-a-chip system.

The evaluation of nanoparticles (NPs) cytotoxicity is crucial for advancing nanotechnology and assessing environmental pollution. However, existing methods for NPs cytotoxicity evaluation suffer from limited accuracy and inadequate information content. In the study, we developed a novel detection platform that enables the identification of cellular carbonyl metabolites at the organ level. The platform is integrated with a cell co-culture lung organ chip (LOC) and a micropillar concentrator. Notably, our work represents the successful measurement of the amounts of cellular metabolites on LOC system. The volatile carbonyl metabolites (VCMs) generated by cells exposure to various types of NPs with different concentrations were captured and detected by high-resolution mass spectrometry (MS). Compared with conventional cell viability and reactive oxygen species (ROS) analysis, our method discerns the toxicological impact of NPs at low concentrations by analyzed VCM at levels as low as ppb level. The LOC system based metabolic gas detection confirmed that low concentrations of NPs have a toxic effect on the cell model, which was not reflected in the fluorescence detection, and the effect of NP material is more significant than the size effect. Furthermore, this method can distinguish different NPs acting on cell models through cluster analysis of multiple VCMs.

Real-time measurement of the trans-epithelial electrical resistance in an organ-on-a-chip during cell proliferation.

The trans-epithelial electrical resistance (TEER) is widely used to quantitatively evaluate cellular barrier function at the organ level in vitro. The measurement of the TEER in organ-on-chips (organ chips) plays a significant role in medical and pharmacological research. However, due to the limitation of the electrical equivalent model for organ chips, the existing TEER measurements usually neglect the changes of the TEER during cell proliferation, resulting in the low accuracy of the measurements. Here, we proposed a new whole-region model of the TEER and developed a real-time TEER measurement system that contains an organ chip with a plate electrode. A whole region circuit model considering the impedance of the non-cell covered region was also established, which enables TEER measurements to be independent of the changes in the cell covered region. The impedance of the non-cell covered region is here attributed to the resistance of the porous membrane. By combining the real-time measurement system and the whole region model, subtle changes in cellular activity during the proliferation stage were measured continuously every 6 minutes and a more sensitive TEER response was obtained. Furthermore, the TEER measurement accuracy was also verified by the real-time measurement of the TEER with stimulation using the permeability enhancer ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). The obtained results indicated that the new proposed whole region model and the real-time measurement system have higher accuracy and greater sensitivity than the traditional model.

Probing electron-atom collision dynamics in gas plasma by high-order harmonic spectroscopy.

Plasma is a complex system involving diverse collisional processes and interactions, such as electron-impact excitation, ionization, recombination, etc. One of the most important methods for studying the properties and dynamics of plasma is to analyze the radiations from plasma. Here, we demonstrate the high-order harmonic (HH) spectroscopy for probing the complex electron-atom collision (EAC) dynamics in a laser-induced gas plasma. These measurements were carried out by using an elliptically polarized pump and a time-delayed linearly polarized probe. The HH spectra from argon and krypton plasmas were recorded by scanning the time delay up to hundreds of picoseconds. We found that the delay-dependent HH yield contains three distinct regions, i.e., the first enhancement, the subsequent suppression, and the final restoration regions. A qualitative analysis shows that these features are clear signatures of the EAC processes and interactions involved in the delay-dependent HH spectroscopy.

Collisional dynamics in laser-induced plasmas: evidence for electron-impact excitation.

We have experimentally investigated the collisional dynamics in femtosecond-laser-induced plasmas and presented the evidence for electron-impact excitation through enhanced high-order harmonic (HH) generation. The measurements were carried out by using an elliptically polarized pump pulse to induce the underdense plasmas and by using a time-delayed linearly polarized probe pulse to drive the HH generation from the plasmas. We found that the rise time of this enhanced HH generation was insensitive to the ellipticity degree (ED) of pump pulse but sensitive to its laser intensity (LI). With further comparison between physical scenarios and qualitative analysis, we demonstrated that the atomic excitation causing the HH enhancement should be attributed to the electron-impact excitation, i.e., the excitation from the collision between neutral atoms and electrons during the lifetime of the underdense plasma.

Ultrafast molecular orbital imaging based on attosecond photoelectron diffraction.

We present ab initio numerical study of ultrafast ionization dynamics of H(2)(+) as well as CO(2) and N(2) exposed to linearly polarized attosecond extreme ultraviolet pulses. When the molecules are aligned perpendicular to laser polarization direction, photonionization of these molecules show clear and distinguishing diffraction patterns in molecular attosecond photoelectron momentum distributions. The internuclear distances of the molecules are related to the position of the associated diffraction patterns, which can be determined with high accuracy. Moreover, the relative heights of the diffraction fringes contain fruitful information of the molecular orbital structures. We show that the diffraction spectra can be well produced using the two-center interference model. By adopting a simple inversion algorithm which takes into account the symmetry of the initial molecular orbital, we can retrieve the molecular orbital from which the electron is ionized. Our results offer possibility for imaging of molecular structure and orbitals by performing molecular attosecond photoelectron diffraction.

Probing rotational wave-packet dynamics with the structural minimum in high-order harmonic spectra.

We investigate the alignment-dependent high-order harmonic spectrum generated from nonadiabatically aligned molecules around the first half rotational revival. It is found that the evolution of the molecular alignment is encoded in the structural minima. To reveal the relation between the molecular alignment and the structural minimum in the high-order harmonic spectrum, we perform an analysis based on the two-center interference model. Our analysis shows that the structural minimum position depends linearly on the inverse of the alignment parameter . This linear relation indicates the possibility of probing the rotational wave-packet dynamics by measuring the spectral minima.

Quantum-orbit analysis for yield and ellipticity of high order harmonic generation with elliptically polarized laser field.

We perform a quantum-orbit analysis for the dependence of high-order-harmonic yield on the driving field ellipticity and the polarization properties of the generated high harmonics. The electron trajectories responsible for the emission of particular harmonics are identified. It is found that, in elliptically polarized driving field, the electrons have ellipticity-dependent initial velocities, which lead to the decrease of the ionization rate. Thus the harmonic yield steeply decreases with laser ellipticity. Besides, we show that the polarization properties of the harmonics are related to the complex momenta of the electron. The physical origin of the harmonic ellipticity is interpreted as the consequence of quantum-mechanical uncertainty of the electron momentum. Our results are verified with the experimental results as well as the numerical solutions of the time dependent Schrödinger equation from the literature.

Influence of large permanent dipoles on molecular orbital tomography.

The influence of large permanent dipoles on molecular orbital tomography via high-order harmonic generation (HHG) is investigated in this work. It is found that, owing to the modification of the angle-dependent ionization rate resulting from the Stark shift, the one-side-recollision condition for the tomographic imaging can not be satisfied even with the few-cycle driving pulses. To overcome this problem, we employ a tailored driving pulse by adding a weak low-frequency pulse to the few-cycle laser pulse to control the HHG process and the recollision of the continuum electrons are effectively restricted to only one side of the core. Then we carried out the orbital reconstruction in both the length and velocity forms. The results show that, the orbital structure can only be successfully reproduced by using the dipole matrix elements projected perpendicular to the permanent dipole in both forms.

Tomographic imaging of asymmetric molecular orbitals with a two-color multicycle laser field.

We theoretically demonstrate a scheme for tomographic reconstruction of asymmetric molecular orbitals based on high-order harmonic generation with a two-color multicycle laser field. It is shown that by adjusting the relative phase of the two fields, the returning electrons can be forced to recollide from one direction for all the orientations of molecules. Thus, the reconstruction of the asymmetric orbitals can be carried out with multicycle laser field. This releases the stringent requirement of a single-cycle pulse with a stabilized and controllable carrier-envelope phase for the tomographic imaging of asymmetric molecular orbitals.

Imprints of the molecular-orbital geometry on the high-harmonic ellipticity.

The influence of the orbital symmetry on the ellipticity of the high-order harmonics is investigated. It is found that the ellipticity maps have distinct shapes for the molecular orbitals with different symmetry. Our analysis shows that the feature of the harmonic ellipticity map is essentially determined by the nodal structure of the nonsymmetric orbital. The results indicate that the molecular-orbital geometry is imprinted on the ellipticity of the high-order harmonics, which invites the use of ellipticity measurements as a probe of the orbital structure for polar molecules.

Broadband large-ellipticity harmonic generation with polar molecules.

We investigate the polarization properties of high harmonic generation from polar molecules with a linearly polarized field. It is found that elliptically polarized harmonics are observed in a wide spectral range from the plateau to the cutoff. Further analyses show that the nonsymmetric structure of the highest occupied molecular orbital is the origin of ellipticity of the harmonics. The results provide a method for generation of large-ellipticity XUV pulses, which will benefit the application of HHG as a tool of detection in materials and biology science.