We present a new type of surface-enhanced Raman scattering (SERS) substrate that exhibits extremely large and uniform cross-section enhancements over a macroscopic (greater than 25 mm2) area. The substrates are fabricated using a femtosecond laser nanostructuring process, followed by thermal deposition of silver. SERS signals from adsorbed molecules show a spatially uniform enhancement factor of approximately 10(7). Spectroscopic characterization of these substrates suggests their potential for use in few or single-molecule Raman spectroscopy.

Fast and selective isolation of single cells with unique spatial and morphological traits remains a technical challenge. Here, we address this by establishing high-speed image-enabled cell sorting (ICS), which records multicolor fluorescence images and sorts cells based on measurements from image data at speeds up to 15,000 events per second. We show that ICS quantifies cell morphology and localization of labeled proteins and increases the resolution of cell cycle analyses by separating mitotic stages. We combine ICS with CRISPR-pooled screens to identify regulators of the nuclear factor κB (NF-κB) pathway, enabling the completion of genome-wide image-based screens in about 9 hours of run time. By assessing complex cellular phenotypes, ICS substantially expands the phenotypic space accessible to cell-sorting applications and pooled genetic screening.

Fluorescence imaging is the most widely used method for unveiling the molecular composition of biological specimens. However, the weak optical emission of fluorescent probes and the tradeoff between imaging speed and sensitivity 1 is problematic for acquiring blur-free images of fast phenomena, such as sub-millisecond biochemical dynamics in live cells and tissues 2 , and cells flowing at high speed 3 . We report a solution that achieves real-time pixel readout rates one order of magnitude faster than a modern electron multiplier charge coupled device (EMCCD) -the gold standard in high-speed fluorescence imaging technology 4 . Deemed fluorescence imaging using radiofrequency-multiplexed excitation (FIRE), this approach maps the image into the radiofrequency spectrum using the beating of digitally synthesized optical fields. We demonstrate diffraction-limited confocal fluorescence imaging of stationary cells at a frame rate of 4.4 kHz, as well as fluorescence microscopy in flow at a throughput of approximately 50,000 cells per second.

The absolute Raman scattering cross section (σ RS ) for the 1584-cm −1 band of benzenethiol at 897 nm (1.383 eV) has been measured to be 8.9 ± 1.8 × 10 −30 cm 2 using a 785-nm pump laser. A temperature-controlled, small-cavity blackbody source was used to calibrate the signal output of the Raman spectrometer. We also measured the absolute surface-enhanced Raman scattering cross section (σ SERS ) of benzenethiol adsorbed onto a silver-coated, femtosecond laser-nanostructured substrate. Using the measured values of 8.9 ± 1.8 × 10 −30 and 6.6 ± 1.3 × 10 −24 cm 2 for σ RS and σ SERS respectively, we calculate an average cross-section enhancement factor (EF) of 0.8 ± 0.3 × 10 6 .

We present a method for improving femtomole-level trace detection (10(9) molecules) using large-area surface-enhanced Raman scattering (SERS) substrates. Using multiphoton-induced exposure of a commercial photoresist, we physically limit the available molecular adsorption sites to only the electromagnetic "hot spots" on the substrate. This process prevents molecules from adsorbing to sites of weak SERS enhancement, while permitting adsorption to sites of extraordinary SERS enhancement. For a randomly adsorbed submonolayer of benzenethiol molecules the average Raman scattering cross section of the processed sample is 27 times larger than that of an unprocessed SERS substrate.

The ability to control chromatic dispersion is paramount in applications where the optical pulsewidth is critical, such as chirped pulse amplification and fiber optic communications. Typically, devices used to generate large amounts (>100 ps/nm) of chromatic dispersion are based on diffraction gratings, chirped fiber Bragg gratings, or dispersion compensating fiber. Unfortunately, these dispersive elements suffer from one or more of the following restrictions: (i) limited operational bandwidth, (ii) limited total dispersion, (iii) low peak power handling, or (iv) large spatial footprint. Here, we introduce a new type of tunable dispersive device, which overcomes these limitations by leveraging the large modal dispersion of a multimode waveguide in combination with the angular dispersion of diffraction gratings to create chromatic dispersion. We characterize the device's dispersion, and demonstrate its ability to stretch a sub-picosecond optical pulse to nearly 2 nanoseconds in 20 meters of multimode optical fiber. Using this device, we also demonstrate single-shot, time-wavelength atomic absorption spectroscopy at a repetition rate of 90.8 MHz.

Porous silicon nanoparticles (PSiNPs) are attractive carriers for targeted drug delivery in nanomedicine. For in vivo applications, the biodegradation property of PSiNPs provides a pathway for their safe clearance from the body. Particles sizes of 80-120 nm are of particular interest as they are important for cellular applications, such as drug delivery for cancer therapy, because these nanoparticles can take advantage of the enhanced permeability and retention effect to deliver drug preferentially to tumors with leaky vasculature, yet large enough to avoid renal clearance. However, the biodegradability rate of such particles is often too fast, which limits particle half-life and potentially reduces their in vivo delivery efficiency. In this work, we focus on the degradation of nanoscale particles and study the effect of both thermal oxidation and silica coating on the stability of PSiNPs in phosphate buffered saline solution (a close mimic of a basic biological fluid). Using thermal oxidation, the half-life of PSiNPs can be varied from 10 min up to 3 h. Using silica coating, the half-life can be extended further to 8 h. The particles produced using both these techniques can be functionalized using standard silica surface chemistries developed for applications in drug delivery.

Improving the efficiency, cell survival, and throughput of methods to modify and control the genetic expression of cells is of great benefit to biology and medicine. We investigate, both computationally and experimentally, a nanostructured substrate made of tipless pyramids for plasmonic-induced transfection. By optimizing the geometrical parameters for an excitation wavelength of 800 nm, we demonstrate a 100-fold intensity enhancement of the electric near field at the cell-substrate contact area, while the low absorption typical for gold is maintained. We demonstrate that such a substrate can induce transient poration of cells by a purely optically induced process.