Simultaneous spatial frequency modulation imaging and micromachining with a femtosecond laser.

A Ti:Al2O3 chirped-pulse amplification system is used to simultaneously image and machine. By combining simultaneous spatial and temporal focusing (SSTF) with spatial frequency modulation for imaging (SPIFI), we are able to decouple the imaging and cutting beams to attain a resolution and a field-of-view that is independent of the cutting beam, while maintaining single-element detection. This setup allows for real-time feedback with the potential for simultaneous nonlinear imaging and imaging through scattering media. The novel SSTF machining platform uses refractive optics that, in general, are prohibitive for energetic, amplified pulses that might otherwise compromise the integrity of the focus as a result of nonlinear effects.

Submillisecond second harmonic holographic imaging of biological specimens in three dimensions.

Optical microscopy has played a critical role for discovery in biomedical sciences since Hooke's introduction of the compound microscope. Recent years have witnessed explosive growth in optical microscopy tools and techniques. Information in microscopy is garnered through contrast mechanisms, usually absorption, scattering, or phase shifts introduced by spatial structure in the sample. The emergence of nonlinear optical contrast mechanisms reveals new information from biological specimens. However, the intensity dependence of nonlinear interactions leads to weak signals, preventing the observation of high-speed dynamics in the 3D context of biological samples. Here, we show that for second harmonic generation imaging, we can increase the 3D volume imaging speed from sub-Hertz speeds to rates in excess of 1,500 volumes imaged per second. This transformational capability is possible by exploiting coherent scattering of second harmonic light from an entire specimen volume, enabling new observational capabilities in biological systems.

Two-dimensional single-pixel imaging by cascaded orthogonal line spatial modulation.

Two-dimensional (2D) images are taken using a single-pixel detector by temporally multiplexing spatial frequency projections from orthogonal, time varying spatial line modulation gratings. Unique temporal frequencies are applied to each point in 2D space, applying a continuous spread of frequencies to one dimension, and an offset frequency applied to each line in the orthogonal dimension. The object contrast information can then be recovered from the electronic spectrum of the single pixel, and through simple processing be reformed into a spatial image.

Plane wave analysis of coherent holographic image reconstruction by phase transfer (CHIRPT).

Fluorescent imaging plays a critical role in a myriad of scientific endeavors, particularly in the biological sciences. Three-dimensional imaging of fluorescent intensity often requires serial data acquisition, that is, voxel-by-voxel collection of fluorescent light emitted throughout the specimen with a nonimaging single-element detector. While nonimaging fluorescence detection offers some measure of scattering robustness, the rate at which dynamic specimens can be imaged is severely limited. Other fluorescent imaging techniques utilize imaging detection to enhance collection rates. A notable example is light-sheet fluorescence microscopy, also known as selective-plane illumination microscopy, which illuminates a large region within the specimen and collects emitted fluorescent light at an angle either perpendicular or oblique to the illumination light sheet. Unfortunately, scattering of the emitted fluorescent light can cause blurring of the collected images in highly turbid biological media. We recently introduced an imaging technique called coherent holographic image reconstruction by phase transfer (CHIRPT) that combines light-sheet-like illumination with nonimaging fluorescent light detection. By combining the speed of light-sheet illumination with the scattering robustness of nonimaging detection, CHIRPT is poised to have a dramatic impact on biological imaging, particularly for in vivo preparations. Here we present the mathematical formalism for CHIRPT imaging under spatially coherent illumination and present experimental data that verifies the theoretical model.

Two-dimensional spatial-frequency-modulated imaging through parallel acquisition of line images.

This Letter demonstrates a two-dimensional imaging technique that uses a line scan camera to resolve one spatial dimension and temporal modulation to resolve the perpendicular dimension. A temporal intensity modulation, which increases linearly in frequency along one direction is applied to an illumination beam. The modulated light distribution is imaged onto an object then onto a line scan camera oriented perpendicularly to the direction of the modulation sweep. A line diffuser is placed shortly before the line scan camera and diffuses light along the direction of modulation so that each pixel collects all modulation frequencies.

Hilbert reconstruction of phase-shifted second-harmonic holographic images.

New techniques are presented that make phase-shifting holography viable for second-harmonic generation (SHG) holography with weak object fields. We developed an intrinsic phase shift calibration of SHG holograms, an algorithm that extracts the reference and object intensity directly from a set of phase-shifted holographic data, and a more robust phase-shifting holography reconstruction algorithm based on π-shifted hologram pairs that permits self-calibration of the phase shift and recovery of the complex field through a Hilbert transform.

Theory of diffraction effects in spatial frequency-modulated imaging.

An analytic theory describing the effects of diffraction and aberrations on single-pixel imaging performed by temporally modulating illumination light is presented. This method encodes spatial information using sinusoidal temporal modulations that are chirped in frequency across the extent of an illumination line focus. With some approximations, a point spread function relationship as a function of defocus or other aberrations is found for both spatially coherent and incoherent cases. The theory is validated through experiments and simulations, including measurement of the transverse and longitudinal optical transfer function, and confirmation of insensitivity to aberrations and significant optical scattering after encoding of spatial information through temporal modulation.

Spatially-chirped modulation imaging of absorbtion and fluorescent objects on single-element optical detector.

Line imaging of fluorescent and absorptive objects with a single-pixel imaging technique that acquires one-dimensional cross-sections through a sample by imposing a spatially-varying amplitude modulation on the probing beam is demonstrated. The fluorophore concentration or absorber distribution of the sample is directly mapped to modulation frequency components of the spatially-integrated temporal signal. Time-domain signals are obtained from a single photodiode, with object spatial frequency correlation encoded in time-domain bursts in the electronic signal from the photodiode.

Subpicosecond fiber-based soliton-tuned mid-infrared source in the 9.7-14.9 microm wavelength region.

We present a fiber-based mid-IR source of ultrafast laser pulses tunable in the 9.7-14.9 microm spectral region based on difference-frequency mixing between an Er:fiber laser and a first-order frequency-shifted soliton pulse. We have measured subpicosecond pulses at a repetition rate of 37 MHz, with a maximum power of 1.5 microW.

Highly achromatic Fourier-transform spectrometer.

We present a simplified all-reflective Fourier transform spectrometer with a split-mirror configuration for use over a broad spectral range with spatially coherent sources. The device is particularly well suited for measurement of broadband laser-like light, with resolution limited by beam size and collimation. Spectra are taken in the near-UV and the mid-IR, a total span of 4.6 octaves, including an octave spanning spectrum. Potential sources of error are investigated both theoretically and experimentally.

Measurement of orientation and susceptibility ratios using a polarization-resolved second-harmonic generation holographic microscope.

Three-dimensional second-harmonic fields, sample orientation, and susceptibility ratios of biological samples are measured using polarization-resolved second-harmonic generation (SHG) microscopy. The three-dimensional (3D) polarization is gathered by measurement of a series of holograms for which excitation and analyzer polarizations are systematically varied, and the 3D SHG field is recovered through numerical back propagation. Harmonophore orientation is resolved in 3D from a sub-set of polarization-resolved SHG holograms. We further expand on previous approaches for the determination of susceptibility ratios, adding the calculation of multiple ratio values to allow intrinsic verification.

Eliminating the scattering ambiguity in multifocal, multimodal, multiphoton imaging systems.

In this work we present how to entirely remove the scattering ambiguity present in existing multiphoton multifocal systems. This is achieved through the development and implementation of single-element detection systems that incorporate high-speed photon-counting electronics. These systems can be used to image entire volumes in the time it takes to perform a single transverse scan (four depths simultaneously at a rate of 30 Hz). In addition, this capability is further exploited to accomplish single-element detection of multiple modalities (two photon excited fluorescence and second harmonic generation) and to perform efficient image deconvolution. Finally, we demonstrate a new system that promises to significantly simplify this promising technology.