Daniel Palanker, Postdoctoral Faculty Sponsor
Neurons undergo nanometer-scale deformations during action potentials, and the underlying mechanism has been actively debated for decades. Previous observations were limited to a single spot or the cell boundary, while movement across the entire neuron during the action potential remained unclear. Here we report full-field imaging of cellular deformations accompanying the action potential in mammalian neuron somas (-1.8 to 1.4 nm) and neurites (-0.7 to 0.9 nm), using high-speed quantitative phase imaging with a temporal resolution of 0.1 ms and an optical path length sensitivity of <4 pm per pixel. The spike-triggered average, synchronized to electrical recording, demonstrates that the time course of the optical phase changes closely matches the dynamics of the electrical signal. Utilizing the spatial and temporal correlations of the phase signals across the cell, we enhance the detection and segmentation of spiking cells compared to the shot-noise-limited performance of single pixels. Using three-dimensional (3D) cellular morphology extracted via confocal microscopy, we demonstrate that the voltage-dependent changes in the membrane tension induced by ionic repulsion can explain the magnitude, time course, and spatial features of the phase imaging. Our full-field observations of the spike-induced deformations shed light upon the electromechanical coupling mechanism in electrogenic cells and open the door to noninvasive label-free imaging of neural signaling.
View details for DOI 10.1073/pnas.1920039117
View details for PubMedID 32341158
Optical phase changes induced by transient perturbations provide a sensitive measure of material properties. We demonstrate the high sensitivity and speed of such methods, using two interferometric techniques: quantitative phase imaging (QPI) in transmission and phase-resolved optical coherence tomography (OCT) in reflection. Shot-noise-limited QPI can resolve energy deposition of about 3.4 mJ/cm2 in a single pulse, which corresponds to 0.8 °C temperature rise in a single cell. OCT can detect deposition of 24 mJ/cm2 energy between two scattering interfaces producing signals with about 30-dB signal-to-noise ratio (SNR), and 4.7 mJ/cm2 when SNR is 45 dB. Both techniques can image thermal changes within the thermal confinement time, which enables accurate single-shot mapping of absorption coefficients even in highly scattering samples, as well as electrical conductivity and many other material properties in biological samples at cellular scale. Integration of the phase changes along the beam path helps increase sensitivity, and the signal relaxation time reveals the size of hidden objects. These methods may enable multiple applications, ranging from temperature-controlled retinal laser therapy or gene expression to mapping electric current density and characterization of semiconductor devices with rapid pump-probe measurements.
View details for PubMedID 29483276
View details for Web of Science ID 000554495702134
View details for Web of Science ID 000554495703198
Photoreceptors initiate vision by converting photons to electrical activity. The onset of the phototransduction cascade is marked by the isomerization of photopigments upon light capture. We revealed that the onset of phototransduction is accompanied by a rapid (<5 ms), nanometer-scale electromechanical deformation in individual human cone photoreceptors. Characterizing this biophysical phenomenon associated with phototransduction in vivo was enabled by high-speed phase-resolved optical coherence tomography in a line-field configuration that allowed sufficient spatiotemporal resolution to visualize the nanometer/millisecond-scale light-induced shape change in photoreceptors. The deformation was explained as the optical manifestation of electrical activity, caused due to rapid charge displacement following isomerization, resulting in changes of electrical potential and surface tension within the photoreceptor disc membranes. These all-optical recordings of light-induced activity in the human retina constitute an optoretinogram and hold remarkable potential to reveal the biophysical correlates of neural activity in health and disease.
View details for DOI 10.1126/sciadv.abc1124
View details for PubMedID 32917686
Real-time quantitative phase imaging has tremendous potential in investigating live biological specimens in vitro. Here we report on a wideband sensitivity-enhanced interferometric microscopy for quantitative phase imaging in real time by employing two quadriwave lateral shearing interferometers based on randomly encoded hybrid gratings with different lateral shears. Theoretical framework to analyze the measurement sensitivity is firstly proposed, from which the optimal lateral shear pair for sensitivity enhancement is also derived. To accelerate the phase retrieval algorithm for real-time visualization, we develop a fully vectorized path-independent differential leveling phase unwrapping algorithm ready for parallel computing, and the framerate for retrieving the phase from each pair of two 4 mega pixel interferograms is able to reach 47.85 frames per second. Experiment results demonstrate that the wideband sensitivity-enhanced interferometric microscopy is capable of eliminating all the periodical error caused by spectral leaking problem and reducing the temporal standard deviation to the half level compared with phase directly retrieved by the interferogram. Due to its high adaptability, the wideband sensitivity-enhanced interferometric microscopy is promising in retrofitting existing microscopes to quantitative phase microscopes with high measurement precision and real-time visualization.
View details for DOI 10.1038/s41598-017-00053-7
View details for PubMedID 28148959
A general lateral shearing interferometry method to measure the wavefront aberrations with a continuously variable shear ratio by the randomly encoded hybrid grating (REHG) is proposed. The REHG consists of a randomly encoded binary amplitude grating and a phase chessboard. Its Fraunhofer diffractions contain only four orders which are the ±1 orders in two orthogonal directions due to the combined modulation of the amplitude and phase. As a result, no orders selection mask is needed for the REHG and the shear ratio is continuously variable, which is beneficial to the variation of sensitivity and testing range for different requirements. To determine the fabrication tolerance of this hybrid grating, the analysis of the effects of different errors on the diffraction intensity distributions is carried out. Experiments have shown that the testing method can achieve a continuously variable shear ratio with the same REHG, and the comparison with a ZYGO GPI interferometer exhibits that the aberration testing method by the REHG is highly precise and also has a good repeatability. This testing method by the REHG is available for general use in testing the aberrations of different optical systems in situ.
View details for DOI 10.1364/AO.54.008913
View details for Web of Science ID 000363311400015
View details for PubMedID 26560379
A compact quadriwave lateral shearing interferometer (QWLSI) with strong adaptability and high precision is proposed based on a novel randomly encoded hybrid grating (REHG). By performing the inverse Fourier transform of the desired ±1 Fraunhofer diffraction orders, the amplitude and phase distributions of the ideally calculated quadriwave grating can be obtained. Then a phase chessboard is introduced to generate the same phase distribution, while the amplitude distribution can be achieved using the randomly encoding method by quantizing the radiant flux on the ideal quadriwave grating. As the Faunhofer diffraction of the REHG only contains the ±1 orders, no order selection mask is ever needed for the REHG-LSI. The simulations and the experiments show that the REHG-LSI exhibits strong adaptability, nice repeatability, and high precision.
View details for DOI 10.1364/OL.40.002245
View details for Web of Science ID 000354708300023
View details for PubMedID 26393710
A common-path and compact wavefront diagnosis system for both continuous and transient wavefronts measurement is proposed based on cross grating lateral shearing interferometer (CGLSI). Derived from the basic CGLSI configuration, this system employs an aplanatic lens to convert the wavefront under test into a convergent beam, which makes it possible for CGLSI to test the wavefront of collimated beams. A geometrical optics model for grating pitch determination and a Fresnel diffraction model for order selection mask design are presented. Then a detailed analysis about the influence of the grating pitch, the distance from the cross grating to the order selection mask and the numerical aperture of the aplanatic lens on the system error is made, and a calibration method is proposed to eliminate the system error. In addition, the differential Zernike polynomials fitting method is introduced for wavefront retrieval. Before our experiment, we have designed several grating pitches and their corresponding order selection mask parameters. In the final comparative experiment with ZYGO interferometer, the wavefront diagnosis system exhibits both high precision and repeatability.
View details for DOI 10.1364/AO.53.007144
View details for Web of Science ID 000343918300027
View details for PubMedID 25402805
An off-axis cyclic radial shearing interferometer (OCRSI) to test a centrally blocked transient wavefront is proposed. Based on the standard cyclic radial shearing interferometer (CRSI), the OCRSI consists of a beam splitter, two folding mirrors, and a Galilean telescope. With the same but reversal tilt introduced to the two mirrors in OCRSI, the shearing interferogram can be obtained even when the central part of the test aperture is blocked. An improved wavefront retrieval method for OCRSI is employed, and a method to obtain the laterally sheared amount between the contracted and expanded beams is proposed. Numerical simulation and comparison experiments with a ZYGO GPI interferometer demonstrate that the OCRSI exhibits high precision and nice repeatability.
View details for DOI 10.1364/OL.38.002493
View details for Web of Science ID 000321770900040
View details for PubMedID 23939091