Two-photon live imaging of direct glia-to-neuron conversion in the mouse cortex

Over the past decade,a growing number of studies have reported transcription factor-based in situ reprogramming that can directly conve rt endogenous glial cells into functional neurons as an alternative approach for n euro regeneration in the adult mammalian central ne rvous system.Howeve r,many questions remain regarding how a terminally differentiated glial cell can transform into a delicate neuron that forms part of the intricate brain circuitry.In addition,concerns have recently been raised around the absence of astrocyte-to-neuron conversion in astrocytic lineage-tra cing mice.In this study,we employed repetitive two-photon imaging to continuously capture the in situ astrocyte-to-neuron conversion process following ecto pic expression of the neural transcription factor NeuroD1 in both prolife rating reactive astrocytes and lineage-tra ced astrocytes in the mouse cortex.Time-lapse imaging over several wee ks revealed the ste p-by-step transition from a typical astrocyte with numero us short,tapered branches to a typical neuro n with a few long neurites and dynamic growth cones that actively explored the local environment.In addition,these lineage-converting cells were able to migrate ra dially or to ngentially to relocate to suitable positions.Furthermore,two-photon Ca2+imaging and patch-clamp recordings confirmed that the newly generated neuro ns exhibited synchronous calcium signals,repetitive action potentials,and spontaneous synaptic responses,suggesting that they had made functional synaptic connections within local neural circuits.In conclusion,we directly visualized the step-by-step lineage conversion process from astrocytes to functional neurons in vivo and unambiguously demonstrated that adult mammalian brains are highly plastic with respect to their potential for neuro regeneration and neural circuit reconstruction.

Two-photon polymerization lithography for imaging optics

Optical imaging systems have greatly extended human visual capabilities,enabling the observation and understanding of diverse phenomena.Imaging technologies span a broad spectrum of wavelengths from x-ray to radio frequencies and impact research activities and our daily lives.Traditional glass lenses are fabricated through a series of complex processes,while polymers offer versatility and ease of production.However,modern applications often require complex lens assemblies,driving the need for miniaturization and advanced designs with micro-and nanoscale features to surpass the capabilities of traditional fabrication methods.Three-dimensional(3D)printing,or additive manufacturing,presents a solution to these challenges with benefits of rapid prototyping,customized geometries,and efficient production,particularly suited for miniaturized optical imaging devices.Various 3D printing methods have demonstrated advantages over traditional counterparts,yet challenges remain in achieving nanoscale resolutions.Two-photon polymerization lithography(TPL),a nanoscale 3D printing technique,enables the fabrication of intricate structures beyond the optical diffraction limit via the nonlinear process of two-photon absorption within liquid resin.It offers unprecedented abilities,e.g.alignment-free fabrication,micro-and nanoscale capabilities,and rapid prototyping of almost arbitrary complex 3D nanostructures.In this review,we emphasize the importance of the criteria for optical performance evaluation of imaging devices,discuss material properties relevant to TPL,fabrication techniques,and highlight the application of TPL in optical imaging.As the first panoramic review on this topic,it will equip researchers with foundational knowledge and recent advancements of TPL for imaging optics,promoting a deeper understanding of the field.By leveraging on its high-resolution capability,extensive material range,and true 3D processing,alongside advances in materials,fabrication,and design,we envisage disruptive solutions to current challenges and a promising incorporation of TPL in future optical imaging applications.

Temperature-feedback two-photon-responsive metal-organic frameworks for efficient photothermal therapy

The realization of real-time thermal feedback for monitoring photothermal therapy(PTT)under near-infrared(NIR)light irradiation is of great interest and challenge for antitumor therapy.Herein,by assembling highly efficient photothermal conversion gold nanorods and a temperature-responsive probe((E)-4-(4-(diethylamino)styryl)-1-methylpyridin-1-ium,PyS)within MOF-199,an intelligent nanoplatform(AMPP)was fabricated for simultaneous chemodynamic therapy and NIR light-induced temperature-feedback PTT.The fluorescence intensity and temperature of the PyS probe are linearly related due to the restriction of the rotation of the characteristic monomethine bridge.Moreover,the copper ions resulting from the degradation of MOF-199 in an acidic microenvironment can convert H_(2)O_(2)into•OH,resulting in tumor ablation through a Fenton-like reaction,and this process can be accelerated by increasing the temperature.This study establishes a feasible platform for fabricating highly sensitive temperature sensors for efficient temperature-feedback PTT.

Two-photon absorption of FAPbBr_(3) perovskite nanocrystals

Perovskite nanocrystals(NCs) with high two-photon absorption(TPA) cross-section are of great interest due to their potential applications in three-dimensional optical data storage and multiphoton fluorescence microscopy. Among various perovskite materials, FAPbBr_(3) NCs show a better development prospect due to their excellent stability. However, there are few reports on their nonlinear optical properties. In this work, the nonlinear optical behavior of FAPbBr_(3) NCs is studied.The methods of multiphoton absorption photoluminescence saturation and open aperture Z-scan technique were applied to determine the TPA cross-section of FAPbBr_(3)NCs, which was around 2.76 × 10^(-45)cm^(4)·s·photon^(-1) at 800 nm. In addition,temperature-dependent photoluminescence induced by TPA was investigated, and the small longitudinal optical phonon energy and electron–phonon coupling strength was obtained, which confirm the weak Pb–Br interaction. Meanwhile, it is found that the exciton binding energy in FAPbBr_(3) NCs was 69.668 me V, which may be ascribed to the strong hydrogen bond interaction. It is expected that our findings will promote the application of FAPbBr_(3) NCs in optoelectronic devices.

A novelα-ketoamide reactivity-based two-photon fluorogenic probe for visualizing peroxynitrite in Parkinson's disease models

Peroxynitrite(ONOO^(-))contributes to oxidative stress and neurodegeneration in Parkinson's disease(PD).Developing a peroxynitrite probe would enable in situ visualization of the overwhelming ONOO^(-)flux and understanding of the ONOO^(-)stress-induced neuropathology of PD.Herein,a novelα-ketoamide-based fluorogenic probe(DFlu)was designed for ONOO^(-)
monitoring in multiple PD models.The results demonstrated that DFlu exhibits a fluorescence turn-on response to ONOO^(-)with high specificity and sensitivity.The efficacy of DFlu for intracellular ONOO^(-)
imaging was demonstrated systematically.The results showed that DFlu can successfully visualize endogenous and exogenous ONOO^(-)in cells derived from chemical and biochemical routes.More importantly,the two-photon excitation ability of DFlu has been well demonstrated by monitoring exogenous/endogenous ONOO^(-)production and scavenging in live zebraflsh PD models.This work provides a reliable and promising
α-ketoamide-based optical tool for identifying variations of ONOO^(-)in PD models.

A simple Bessel module with tunable focal depth and constant resolution for commercial two-photon microscope

The volumetric imaging of two-photon microscopy expands the focal depth and improves the throughput,which has unparalleled superiority for three-dimension samples,especially in neuroscience.However,emerging in volumetric imaging is still largely customized,which limits the integration with commercial two-photon systems.Here,we analyzed the key parameters that modulate the focal depth and lateral resolution of polarized annular imaging and proposed a volumetric imaging module that can be directly integrated into commercial two-photon systems using conventional optical elements.This design incorporates the beam diameter adjustment settings of commercial two-photon systems,allowing flexibility to adjust the depth of focus while maintaining the same lateral resolution.Further,the depth range and lateral resolution of the design were verified,and the imaging throughput was demonstrated by an increase in the number of imaging neurons in the awake mouse cerebral cortex.

Production of χc and ηc in Ultra-Peripheral Collisions with Two-Photon Processes

We calculate the production of χc and ηc by the two-photon process in ultra-peripheral heavy ion collisions at Relativistic Heavy Ion Collider (RHIC) and Large Hadron Collider (LHC) energies. The differential cross section of transverse momentum distribution and rapidity distribution for (H = χc and ηc), are estimated by using the equivalent photon flux in the impact parameter space. The numerical results indicate that the study of χc and ηc in ultra-peripheral heavy ion collisions are feasible at RHIC and LHC energies.

Multi-branched carbazole derivatives for two-photon absorption and two-photon excited fluorescence

Carbazole-core multi-branched chromophores 9-ethyl- 3, 6-bis （ 2- { 4- [ 5- （4-tert-butyl-phenyl） - [ 1, 3, 4 ] oxadiazol-2-yl ] - phenyl }-vinyl） -carbazole（3） and 9-ethyl-3-（ 2- {4-[ 5-（4-tert-butyl- phenyl） -[ 1, 3, 4 ] oxadiazol-2-yl ] -phenyl }-vinyl ） -carbazole （ 2 ） are synthesized through Wittig reaction and characterized by nuclear magnetic resonance（NMR）and infrared（IR）. The two- photon absorption properties of chromophores are investigated. These chromophores exhibit large two-photon absorption crosssections and strong blue two-photon excited fluorescence. The cooperative enhancement of two-photon absorption（TPA） in the multi-branched structures is observed. This enhancement is partly attributed to the electronic coupling between the branches. The electronic push-pull structures in the arm and their cooperative effects help the extended charge transfer for TPA.

Two-photon Excited Fluorescence of Bithiophene Derivatives

Two new bithiophene derivatives named as 5, 5-bis(p-N,N-dimethylaminostyryl)-2, 2 -bithiophene (BMSBT), and 5, 5-bis(p-N,N-diethylaminostyryl)-2, 2-bithiophene (BESBT) have been synthesized. Both compounds can emit strong single-photon excited fluorescence (SPEF) and two-photon excited fluorescence (TPEF) with the emission peaks around ~560 nm and with the lifetime of ~1ns.

Synthesis, Structure of a Symmetrically Substituted Stilbene with Strong Two-photon Absorption and Up-converted Blue Fluorescence

Efficient Ti-catalyzed reductive coupling methodology was first employed to synthesize the symmetrical bis-donor stilbene, trans-4, 4'-bis[diphenyl amino] stilbene (BDPAS). X-ray diffraction analyses reveal that this new crystal belongs to the triclinic crystal system of centro-symmetric P-1 space group. The DBPAS solution, with the linear transmission at wavelength of greater than or equal to 450 nm, possesses large two-photon absorption cross section as high as 39.4x10(-48) cm(4).s/photon resulting in strong two-photon induced blue fluorescence of 460 nm, pumped by 740 nm laser irradiation.

Femtosecond Two-Photon Detachment of Cu^- Studied By Photoelectron Imaging

The wavelength dependence of photoelectron angular distributions （PADs） of two-photon detachment of Cu^- has been directly studied by using the photoelectron map imaging. Results show that for the laser field intensity of 6.0×10^10W/cm^2, PADs exhibit dramatic change with the external field wavelength. Comparison between the experimental observation and the lowest-order perturbation theory prediction indicates that the pattern of PADs can be explained by the interference of the s and d partial waves in the final state. Relative contri- butions of s and d partial waves in the two-photon detachment at different laser wavelengths are obtained.

Synthesis and Two-photon Absorption Properties of s-Triazine Derivatives

Two new s-triazine derivatives, which belong to linear dipolar type and triangle octupolar type respectively, have been synthesized. The structure of the dipolar compound has been determined by X-ray diffraction. The two-photon absorption cross-section σ, and the two-photon excited fluorescence (TPEF) intensities are increased significantly from dipolar compound to octupolar compound.

New Thiophene Derivatives with Two-photon Excited Fluorescence

Two new compounds involving a thiophene moiety named as 2,5-bis[4-(N,N- diphenyl- amino)styryl]thiophene (BPST) and 2,5-bis[4-(N,N-diethylamino)styryl]thiophene (BEST) have been synthesized. The two-photon absorption cross section of BPST was measured as large as 256 × 10-50 cm4·s/photon, when it was excited by 800 nm femtosecond laser.

Optical Rectification Induced by Al-Si Schottky Barrier Potential and Mechanism of Two-Photon Response

By observing two-photon response and anisotropy of the light-induced voltage in Al-Si Schottky barrier potential,it is certified from the experimental and theoretical analysis that the built-in electric field generated by the Schottky barrier potential will induce the phenomena of optical rectification in Si photodiode.Thus,it is deduced that there must be double-frequency absorption caused by phase-mismatch in the mechanism of two-photon response of Si photodiode.If the intensity of the built-in electric field is strong enough,the double-frequency absorption will be the main factor of the two-photon response,which is different from the conventional opinion that the two-photon response is just the two-photon absorption.

A Surface Femtosecond Two-Photon Photoemission Spectrometer for Excited Electron Dynamics and Time-Dependent Photochemical Kinetics

A surface femtosecond two-photon photoemission （2PPE） spectrometer devoted to the study of ultrafast excited electron dynamics and photochemical kinetics on metal and metal oxide surfaces has been constructed. Low energy photoelectrons are measured using a hemispherical electron energy analyzer with an imaging detector that allows us to detect the energy and the angular distributions of the photoelectrons simultaneously. A Mach-Zehnder interferom- eter was built for the time-resolved 2PPE （TR-2PPE） measurement to study ultrafast surface excited electron dynamics, which was demonstrated on the Cu（111） surface. A scheme for measuring time-dependent 2PPE （TD-2PPE） spectra has also been developed for studies of surface photochemistry. This technique has been applied to a preliminary study on the photochemical kinetics on ethanol/TiO2（110）. We have also shown that the ultrafast dynamics of photoinduced surface excited resonances can be investigated in a reliable way by combining the TR-2PPE and TD-2PPE techniques.

Isotope Effects on Two-Photon Population Transfer Processes of HF and DF

The isotope effects of XF （X=H, D） on the population transfer process via two-photon resonance excitation are investigated by solving the time-dependent SchrSdinger equation. The vibrational levels v=0 and 2 of the ground electronic state are taken to be the initial and target states, respectively, for the two molecular systems. The influences of the field peak amplitude and pulse duration on the population transfer process are discussed in detail. The pulse duration is required to be longer than 860 fs for the DF molecule to achieve a relatively high transfer probability （more than 80%）, while the one for the HF molecule is just required to be longer than 460 fs. Moreover, the intermediate level v=1 and the higher level v=3 may play more important roles in the two-photon resonance process for the DF molecule, compared to the roles in the process for the HF molecule.

Theoretical Studies on One-photon and Two-photon Absorption Properties of Pyrene-core Derivatives

The analytic response theory at density functional theory level is applied to investigate onephoton and two-photon absorption properties of a series of recently synthesized pyrene-core derivatives. The theoretical results show that there are a few charge-transfer states for each compound in the lower energy region. The one-photon absorption properties of the five investigated compounds are highly consistent with those given by experimental measurements. The two-photon absorption intensities of the compounds are greatly enhanced with the increments of the molecular sizes, in which the two-photon absorption cross section of the four-branched compound is about 5.6 times of that of the mono-branched molecule. Fhrthermore, it is shown that the two-photon absorption properties are sensitive to the geometrical arrangements.

Measurement Induced Enhancement of Squeezing in Nondegenerate Two-Photon Jaynes-Cummings Model

Squeezing properties in the nondegenerate two-photon Jaynes-Cummings model are investigated. The effects of direct selective atomic measurement and the application of the classical field followed by atomic measurement are analyzed. Different values of the parameters of the classical field are taken into account. It is found that the field squeezing can be enhanced by measurement.

Level crossing in a two-photon Jaynes-Cummings model

In this paper,the energy spectrum of the two-photon Jaynes-Cummings model（TPJCM） is calculated exactly in the non-rotating wave approximation（non-RWA）,and we study the level-crossing problem by means of fidelity.A narrow peak of the fidelity is observed at the level-crossing point,which does not appear at the avoided-crossing point.Therefore fidelity is perfectly suited for detecting the level-crossing point in the energy spectrum.

Populating ^(229m)Th via two-photon electronic bridge mechanism

The isomer ^(229m)Th is the most promising candidate for clocks based on the nuclear transition because it has the lowest excitation energy of only 8.10±0.17 eV.Various experiments and theories have focused on methods of triggering the transition between the ground state and isomeric state,among which the electronic bridge(EB)is one of the most efficient.In this paper,we propose a new electronic bridge mechanism via two-photon excitation based on quantum optics for a two-level nuclear quantum system.The long-lived 7 s1/2 electronic shell state of^(229m)Th^(3+),with a lifetime of approximately 0.6 s,is chosen as the initial state and the atomic shells(7 s-10 s)could be achieved as virtual states in a two-photon process.When the virtual states return to the initial state 7 s1/2,there is a chance of triggering the nucleus 229Th^(3+),to its isomeric state ^(229m)Th ^(3+),via EB.Two lasers at moderate intensity((10^(10)-10^(14))W/m^(2)),with photon energies near the optical range,are expected to populate the isomer at a saturated rate of approximately 10^(9) s^(-1),which is much higher than that due to other mechanisms.We believe that this twophoton EB scheme can help in the development of nuclear clocks and deserves verification via a series of experiments with ordinary lasers in laboratories.