Surface- and bulk-micromachined two-dimensional scanner driven by angular vertical comb actuators

In this paper, we present the design, fabrication, and measurements of a two-dimensional (2-D) optical scanner with electrostatic angular vertical comb (AVC) actuators. The scanner is realized by combining a foundry-based surface-micromachining process (Multi-User MEMS Processes-MUMPs) with a three-mask deep-reactive ion-etching (DRIE) postfabrication process. The surface-micromachining provides versatile mechanical design and electrical interconnect while the bulk micromachining offers high-aspect ratio structures leading to flat mirrors and high-force, large-displacement actuators. The scanner achieves dc mechanical scanning ranges of +/- 6.2 degrees (at 55 Vdc) and +/- 4.1 degrees (at 50 Vdc) for the inner and outer gimbals, respectively. The resonant frequencies are 315 and 144 Hz for the inner and the outer axes, respectively. The 1-mm-diameter mirror has a radius of curvature of over 50 cm

Compact and flexible raster scanning multiphoton endoscope capable of imaging unstained tissue

We present a compact and flexible endoscope (3-mm outer diameter, 4-cm rigid length) that utilizes a miniaturized resonant/nonresonant fiber raster scanner and a multielement gradient-index lens assembly for two-photon excited intrinsic fluorescence and second-harmonic generation imaging of biological tissues. The miniaturized raster scanner is fabricated by mounting a commercial double-clad optical fiber (DCF) onto two piezo bimorphs that are aligned such that their bending axes are perpendicular to each other. Fast lateral scanning of the laser illumination at 4.1 frames/s (512 lines per frame) is achieved by simultaneously driving the DCF cantilever at its resonant frequency in one dimension and nonresonantly in the orthogonal axis. The implementation of a DCF into the scanner enables simultaneous delivery of the femtosecond pulsed 800-nm excitation source and epi-collection of the signal. Our device is able to achieve a field-of-view (FOVxy) of 110 μm by 110 μm with a highly uniform pixel dwell time. The lateral and axial resolutions for two-photon imaging are 0.8 and 10 μm, respectively. The endoscope’s imaging capabilities were demonstrated by imaging ex vivo mouse tissue through the collection of intrinsic fluorescence and second-harmonic signal without the need for staining. The results presented here indicate that our device can be applied in the future to perform minimally invasive in vivo optical biopsies for medical diagnostics

Visually evoked activity in cortical cells imaged in freely moving animals

We describe a miniaturized head-mounted multiphoton microscope and its use for recording Ca2+ transients from the somata of layer 2/3 neurons in the visual cortex of awake, freely moving rats. Images contained up to 20 neurons and were stable enough to record continuously for >5 min per trial and 20 trials per imaging session, even as the animal was running at velocities of up to 0.6 m/s. Neuronal Ca2+ transients were readily detected, and responses to various static visual stimuli were observed during free movement on a running track. Neuronal activity was sparse and increased when the animal swept its gaze across a visual stimulus. Neurons showing preferential activation by specific stimuli were observed in freely moving animals. These results demonstrate that the multiphoton fiberscope is suitable for functional imaging in awake and freely moving animals