Abstract. Super-resolution microscopy techniques-capable of overcoming the diffraction limit of light-have opened new opportunities to explore subcellular structures and dynamics not resolvable in conventional far-field microscopy. However, relying on staining with exogenous fluorescent markers, these techniques can sometimes introduce undesired artifacts to the image, mainly due to large tagging agent sizes and insufficient or variable labeling densities. By contrast, the use of endogenous pigments allows imaging of the intrinsic structures of biological samples with unaltered molecular constituents. Here, we report label-free photoacoustic (PA) nanoscopy, which is exquisitely sensitive to optical absorption, with an 88 nm resolution. At each scanning position, multiple PA signals are successively excited with increasing laser pulse energy. Because of optical saturation or nonlinear thermal expansion, the PA amplitude depends on the nonlinear incident optical fluence. The high-order dependence, quantified by polynomial fitting, provides super-resolution imaging with optical sectioning. PA nanoscopy is capable of super-resolution imaging of either fluorescent or nonfluorescent molecules. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
Optical-resolution photoacoustic microscopy (OR-PAM) has become a major experimental tool of photoacoustic tomography, with unique imaging capabilities for various biological applications. However, conventional imaging systems are all table-top embodiments, which preclude their use in internal organs. In this study, by applying the OR-PAM concept to our recently developed endoscopic technique, called photoacoustic endoscopy (PAE), we created an optical-resolution photoacoustic endomicroscopy (OR-PAEM) system, which enables internal organ imaging with a much finer resolution than conventional acousticresolution PAE systems. OR-PAEM has potential preclinical and clinical applications using either endogenous or exogenous contrast agents.
Picosecond absorption relaxation-central to many disciplines-is typically measured by ultrafast ͑femtosecond or picosecond͒ pump-probe techniques, which however are restricted to optically thin and weakly scattering materials or require artificial sample preparation. Here, we developed a reflection-mode relaxation photoacoustic microscope based on a nanosecond laser and measured picosecond absorption relaxation times. The relaxation times of oxygenated and deoxygenated hemoglobin molecules, both possessing extremely low fluorescence quantum yields, were measured at 576 nm. The added advantages in dispersion susceptibility, laser-wavelength availability, reflection sensing, and expense foster the study of natural-including strongly scattering and nonfluorescent-materials. © 2010 American Institute of Physics. ͓doi:10.1063/1.3500820͔Picosecond absorption relaxation is central to many optical phenomena in physics, chemistry, biology, and medicine-such as absorption saturation, 1-3 photosynthesis, 4 and photolysis. [5][6][7] Typically, relaxation times are studied via an ultrafast ͑femtosecond or picosecond͒ pump-probe technique by measuring either fluorescence in a reflection configuration or absorption in a transmission configuration. 8,9 In the reflection-based approach, the molecules are excited by a laser pulse through the transition ͉0͘ → ͉i͘. A probe pulse with a variable delay time relative to the pump pulse monitors the time evolution of the population density N i ͑t͒. 8 When applied to molecules with low fluorescence quantum yield, this approach is noisy. In the transmission-based approach, the relaxation time is extracted by measuring pump-induced changes in probe-beam transmittivity ⌬T = T͑I p ͒ − T͑0͒, where T͑I p ͒ and T͑0͒ are the probe beam transmittivities with and without a pump of intensity I p . 9 In both approaches, the time resolution is limited by the pulse width of both the pump and the probe pulses. Hence, not only is an expensive picosecond or femtosecond laser required, but also the detection is susceptible to pulse broadening in dispersive media. Alternatively, intensity correlation of nanosecond laser pulses can be used. 10,11 Working in transmission mode, this technique is not suitable for optically thick media. All of the current techniques are limited to optically thin and weakly scattering materials or require artificial sample preparation.Photoacoustic microscopy and photoacoustic computed tomography, usually based on nanosecond laser excitation, are effective functional and molecular imaging tools in vivo. In the photoacoustic phenomenon, light is absorbed by a material and converted to heat. The subsequent thermoelastic expansion generates a detectable acoustic wave. 12 Photoacoustic sensing presents an exquisite sensitivity due to its inherent background-free nature. Its relative sensitivity to absorption reaches the theoretical limit of 100%, by far the highest among all optical imaging modalities. Most quantitative photoacoustic studies have assumed a linear dependence between ...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.