High-throughput multiphoton-induced three-dimensional ablation and imaging for biotissues

Abstract

Abstract:In this study, a temporal focusing-based high-throughput multiphoton-induced ablation system with axially-resolved widefield multiphoton excitation has been successfully applied to rapidly disrupt biotissues. Experimental results demonstrate that this technique features high efficiency for achieving large-area laser ablation without causing serious photothermal damage in non-ablated regions. Furthermore, the rate of tissue processing can reach around 1.6 × 10 6 μm 3 /s in chicken tendon. Moreover, the temporal focusing-based multiphoton system can be efficiently utilized in optical imaging through iterating high-throughput multiphoton-induced ablation machining followed by widefield optical sectioning; hence, it has the potential to obtain molecular images for a whole bio-specimen. References and links1. W. R. Zipfel, R. M. Williams, and W. W. Webb, "Nonlinear magic: multiphoton microscopy in the biosciences," Nat. Biotechnol. 21(11), 1369-1377 (2003). 2. I. Freund and M. Deutsch, "Second-harmonic microscopy of biological tissue," Opt. Lett. 11(2), 94-96 (1986) "Nonlinear structured-illumination enhanced temporal focusing multiphoton excitation microscopy with a digital micromirror device," Biomed.

Temporal focusing‐based widefield multiphoton microscopy with spatially modulated illumination for biotissue imaging

“…The TFMPEM with the breakthrough approach of high‐throughput illumination and detection capability has been applied for fast Brownian motion tracking , widefield fluorescence lifetime imaging , and 3D neuronal activity observation . Further, the fast‐imaging ability of the TFMPEM has been integrated with holographic optical tweezers and a large‐area multiphoton‐induced ablation technique , thereby evolving into a multifunctional system in which both capabilities are realized with the use of only a single objective.…”

Oxygen-Permeable Films for Continuous Additive, Subtractive, and Hybrid Additive/Subtractive Manufacturing

“…[1][2][3] Current fabrication methods based on additive and/or subtractive approaches are limited in their ability to print complex multiscale three-dimensional (3D) constructs and devices using soft materials. [4][5][6][7] Subtractive manufacturing methods such as soft lithography, nanoimprint lithography, electron-beam etching, and ion beam milling lithography can provide highresolution multiscale features, however, the fabrication is mostly limited for developing two-dimensional devices. [8][9][10] Advanced techniques such as multiphoton ablation (MPA) can sculpt 3D features within soft materials, however, limited processing depth and low scalability/throughput make this method unfeasible for making functional devices.…”

Live imaging of contracting muscles with wide-field second harmonic generation microscopy using a high power laser

“…SHG image contrast relies on the frequency-doubled response of molecules to high intensity laser pulses. For optimal SHG imaging, a laser beam from a femtosecond oscillator is focused with a microscope objective lens onto a sample, and the laser beam is raster-scanned to acquire an image either via pixel by pixel signal detection with a single element photodetector (usually a photomultiplier tube or an avalanche photodiode), or a sample is imaged onto an array detector (a charge-coupled device, CCD, or a complementary metal-oxide-semiconductor, CMOS) [6][7][8][9][10]. A large field of view image acquisition rate is limited by the laser scanning speed and SHG efficiency of the sample, restricting the investigations to static or slowly changing (on the order of seconds to minutes) biological structures.…”