Single-molecule localization microscopy (SMLM) provides unmatched high resolution but relies on accurate drift correction due to the long acquisition time for each field of view. A popular drift correction is implemented via referencing to fiducial markers that are assumed to be firmly immobilized and remain stationary relative to the imaged sample. However, there is so far lack of efficient approaches for evaluating other motions except sample drifting of immobilized markers and for addressing their potential impacts on images. Here, we developed a new approach for quantitatively assessing the motions of fiducial markers relative to the sample via mean squared displacement (MSD) analysis. Our findings revealed that over 90% of immobilized fluorescent beads in the SMLM imaging buffer exhibited higher MSDs compared to stationary beads in dry samples and displayed varying degrees of wobbling relative to the imaged field. By excluding extremely high-MSD beads in each field from drift correction, we optimized drift correction and experimentally measured localization precision. In SMLM experiments of cellular microtubules, we also found that including only relatively low-MSD beads for drift correction significantly improved the image resolution and quality. Our study presents a simple and effective approach to assess the potential relative motions of fiducial markers and emphasizes the importance of pre-screening fiducial markers for improved image quality and resolution in SMLM imaging.
We report on an experimental measurement of the pulse front tilt (PFT) of spatiotemporally focused femtosecond laser pulses in the focal plane in both air and bulk transparent materials, which is achieved by examination of the interference pattern between the spatiotemporally focused pulse and a conventional focused reference pulse as a function of time delay between the two pulses. Our simulation results agree well with the experimental observations.
Interferometry/methods , Lasers , Plasma Gases/chemistry , Equipment Design , Materials Testing
We experimentally demonstrate ultrafast dynamic of generation of the 337-nm nitrogen laser by injecting an external seed pulse into a femtosecond laser filament pumped by a circularly polarized laser pulse. In the pump-probe scheme, it is revealed that the population inversion between the C(3)Π(u) and B(3)Π(g) states of N(2) for the free-space 337-nm laser is firstly built up on the timescale of several picoseconds, followed by a relatively slow decay on the timescale of tens of picoseconds, depending on the nitrogen gas pressure. By measuring the intensities of 337-nm signal from nitrogen gas mixed with different concentrations of oxygen gas, it is also found that oxygen molecules have a significant quenching effect on the nitrogen laser signal. Our experimental observations agree with the picture of electron-impact excitation.
We report on experimental realization of impulsive rotational Raman scattering from neutral nitrogen molecules in a femtosecond laser filament using an intense self-induced white-light seeding "air laser" generated during the filamentation of an 800 nm Ti:sapphire laser in nitrogen gas. The impulsive rotational Raman fingerprint signals are observed with a maximum conversion efficiency of â¼0.8%. Our observation provides a promising way of remote identification and location of chemical species in the atmosphere by a rotational Raman scattering of molecules.
We report on experimental examination of two-photon fluorescence excitation (TPFE) at the focus of a spatially chirped femtosecond laser beam, which reveals an unexpected tilted peak intensity distribution in the focal spot. Our theoretical calculation shows that the tilting of the peak intensity distribution originates from the fact that along the optical axis of objective lens, the spatiotemporally focused pulse reaches its shortest duration exactly at the focal plane. However, when moving away from the optical axis along the direction of spatial chirp of the incident pulse, the pulse reaches its shortest duration either before or after the focal plane, depending on whether the pulse duration is measured above or below the optical axis as well as the sign of the spatial chirp. The tilting of the peak intensity distribution in the focal spot of the spatiotemporally focused femtosecond laser beam can play important roles in applications such as femtosecond laser micromachining and bio-imaging.
We present the generation of self-induced-white-light-seeded lasing action of nitrogen molecules in air by a Ti:sapphire femtosecond laser (800 nm, 5.5 mJ) and demonstrate that such lasing action is strongly influenced by external focusing conditions. It is found that the self-seeded lasing signal of N(2)(+) at ~391 nm decreases dramatically by orders of magnitude and ultimately disappears when the focal length of an external lens increases from 0.5 m to 1 m. By using a telescope, it is shown that such limitation can be overcome and the 391 nm lasing can be controlled to occur at remotely designated distance, providing a possibility for practical applications in standoff spectroscopy.
We demonstrate unexpectedly strong second harmonic generation (SHG) in argon gas by use of spatio-temporally focused (SF) femtosecond laser pulses. The resulting SHG by the SF scheme at a 75 cm distance shows a significantly enhanced efficiency than that achieved with conventional focusing schemes, which offers a new promising possibility for standoff applications. Our theoretical calculations reasonably reproduce the experimental observations, which indicate that the observed SHG mainly originates from the gradient of non-uniform plasma dynamically controlled by the SF laser field.
The study of the ionization process of molecules in an intense infrared laser field is of paramount interest in strong-field physics and constitutes the foundation of imaging of molecular valence orbitals and attosecond science. We show measurement of alignment-dependent ionization probabilities of the lower-lying orbitals of the molecules by experimentally detecting the alignment dependence of fluorescence emission from tunnel ionized carbon dioxide molecules. The experimental measurements are compared with the theoretical calculations of the strong field approximation and molecular Ammosov-Delone-Krainov models. Our results demonstrate the feasibility of an all-optical approach for probing the ionization dynamics of lower-lying orbitals of molecules, which is until now still difficult to achieve by other techniques. Moreover, the deviation between the experimental and theoretical results indicates the incompleteness of current theoretical models for describing strong field ionization of molecules.
Recently, amplification of harmonic-seeded radiation generated through femtosecond laser filamentation in air has been observed, giving rise to coherent emissions at wavelengths corresponding to transitions between different vibrational levels of the electronic B(2)Σ(u)(+) and X(2)Σ(g)(+) states of molecular nitrogen ions [Phys. Rev. A. 84, 051802(R) (2011)]. Here, we carry out systematic investigations on its physical mechanism. Our experimental results do not support the speculation that such excellent coherent emissions could originate from nonlinear optical processes such as four-wave mixing or stimulated Raman scattering, leaving stimulated amplification of harmonic seed due to the population inversion generated in molecular nitrogen ions the most likely mechanism.
Air Pollutants/analysis , Air/analysis , Environmental Monitoring/instrumentation , Lasers , Nephelometry and Turbidimetry/instrumentation , Nitrogen/analysis , Equipment Design , Equipment Failure Analysis
We theoretically investigate the selective enhancement of high-harmonic generation (HHG) in a small spectral range when an orthogonal-polarized two-color laser field interacts with aligned O(2) molecules. It is shown clearly that the enhanced narrow-bandwidth emission near the cutoff of the HHG spectrum can be effectively controlled by the molecular alignment angle, laser intensity and the relative phase of two-color laser fields. Furthermore, the strong dependence of narrow-bandwidth HHG on molecular alignment angle indicates that it encodes information about O(2) molecular orbitals, so it may be an alternative method for reconstruction of O(2) molecular orbitals.
Models, Chemical , Oxygen/chemistry , Oxygen/radiation effects , Computer Simulation , Lasers , Molecular Conformation , Scattering, Radiation
We theoretically investigate the X-ray supercontinuum generated by interaction of multi-cycle, chirped polarization gating pulses with the helium gas. It is shown that with this scheme, an isolated sub-50-attosecond pulse can be obtained straightforwardly without any phase compensation. Interestingly, if one selects an extremely broad spectral range near the high-order harmonic cutoff, an isolated and intense sub-24-attosecond pulse can be generated after phase compensation, which could be used to detect and control the electronic dynamics inside the atoms. Furthermore, it is found that the generation of such a broad and smooth X-ray supercontinuum is not so stringent on the selection of the simulated parameters, allowing for the experimental demonstration of this technique in the future.
We investigate theoretically the enhancement of the low-order harmonic emission from a polar molecular medium. The results show that, by using a control laser field, the intensity of the spectral signals near fourth-order harmonics will increase over 25 times as a result of the four-wave mixing process. Moreover, the enhancement effects depend strongly on the carrier-envelope phase of the initial laser fields, which cannot be found in a symmetric system.
Computer Simulation , Lasers , Light , Models, Theoretical , Semiconductors , Signal Processing, Computer-Assisted/instrumentation , Spectrum Analysis/methods , Humans , Quantum Theory
We report on the investigation on harmonic-seeded remote laser emissions at 391 nm wavelength from strong-field ionized nitrogen molecules in three different gas mixtures, i.e., N2-Ar, N2-Xe and N2-Ne. We observed a decrease in the remote laser intensity in the N2-Xe mixture because of the decreased clamped intensity in the filament; whereas in the N2-Ne mixture, the remote laser intensity slightly increases because of the increased clamped intensity within the filament. Remarkably, although the clamped intensity in the filament remains nearly unchanged in the N2-Ar mixture because of the similar ionization potentials of N2 and Ar, a significant enhancement of the lasing emission is realized in the N2-Ar mixture. The enhancement is attributed to the stronger third harmonic seed, and longer gain medium due to the extended filament.
We reported a novel femtosecond-laser-activated fluorescence emission from indole solutions upon excitation by the second harmonic wavelength of a femtosecond oscillator. A new absorption band around 400 nm and corresponding fluorescent band in the green domain were produced after the irradiation of femtosecond laser. This femtosecond-laser-activated luminescence process that allows the use of visible wavelength as a substitute for UV light to excite fluorescence from indole would extend applications based on indole chromophore. Furthermore, the photoactived emission can act as a fluorescence lifetime probe to measure the polarity in complex biological systems since it is polarity-sensitive. High performance liquid chromatography with fluorescence detector (HPLC-FLU) and high performance liquid chromatography with mass spectrometer (HPLC-MS) analysis demonstrate that the origin of the photoactivated fluorescence is new molecular species that generated in indole solution upon femtosecond laser irradiation.
Indoles/chemistry , Oscillometry , Chromatography, High Pressure Liquid , Fluorescence , Lasers , Mass Spectrometry , Photochemical Processes , Solutions , Spectrometry, Fluorescence , Time Factors , Ultraviolet Rays
We report on the fabrication of hollow microfluidic channels with a circular cross-sectional shape embedded in fused silica by spatiotemporally focusing the femtosecond laser beam. We demonstrate both theoretically and experimentally that the spatiotemporal focusing of femtosecond laser beam allows for the creation of a three-dimensionally symmetric spherical intensity distribution at the focal spot.
