. Significance: Deep tissue noninvasive high-resolution imaging with light is challenging due to the high degree of light absorption and scattering in biological tissue. Photoacoustic imaging (PAI) can overcome some of the challenges of pure optical or ultrasound imaging to provide high-resolution deep tissue imaging. However, label-free PAI signals from light absorbing chromophores within the tissue are nonspecific. The use of exogeneous contrast agents (probes) not only enhances the imaging contrast (and imaging depth) but also increases the specificity of PAI by binding only to targeted molecules and often providing signals distinct from the background. Aim: We aim to review the current development and future progression of photoacoustic molecular probes/contrast agents. Approach: First, PAI and the need for using contrast agents are briefly introduced. Then, the recent development of contrast agents in terms of materials used to construct them is discussed. Then, various probes are discussed based on targeting mechanisms, in vivo molecular imaging applications, multimodal uses, and use in theranostic applications. Results: Material combinations are being used to develop highly specific contrast agents. In addition to passive accumulation, probes utilizing activation mechanisms show promise for greater controllability. Several probes also enable concurrent multimodal use with fluorescence, ultrasound, Raman, magnetic resonance imaging, and computed tomography. Finally, targeted probes are also shown to aid localized and molecularly specific photo-induced therapy. Conclusions: The development of contrast agents provides a promising prospect for increased contrast, higher imaging depth, and molecularly specific information. Of note are agents that allow for controlled activation, explore other optical windows, and enable multimodal use to overcome some of the shortcomings of label-free PAI.
Feedback-based wavefront shaping is a promising and versatile technique for enhancing the contrast of a target signal for both coherent and incoherent light through a highly scattering medium. However, this technique can fail for a dynamic sample with a short correlation time. So far, most proposed methods for high-speed wavefront shaping can only directly enhance the intensity of coherent light but not incoherent light. Here we try to fill this gap to directly enhance incoherent light with high speed, such as fluorescence, which is essential in extending wavefront shaping to biomedical applications. For this purpose, we develop a technique based on a single acousto-optic deflector (AOD) with field-programmable gate array (FPGA) acceleration for spatiotemporal focusing within milliseconds. With the digital time gating of the feedback signal, spatiotemporal focusing of laser light with high contrast can be formed behind dynamic scattering media in milliseconds resulting in fluorescence enhancement. Furthermore, FPGA-based wavefront shaping is shown to effectively enhance fluorescence directly behind dynamic samples with short correlation times.
Single pixel spectroscopy based on Hadamard transform (SPS-HT) has been applied widely because of its capability of wavelength multiplexing and associated advantage in signal-to-noise ratio. In this paper, we propose a sequency encoding single pixel spectroscopy (SESPS) based on two-dimensional (2D) masks for concurrent coding of all Hadamard coefficients instead of one-dimensional (1D) Hadamard masks (only coding one coefficient at a time) widely used in the traditional SPS-HT. Moreover, each Hadamard coefficient is coded along the time dimension with a different sequency value such that the alternating current (AC) measurements of the time-domain signal can be used to reconstruct all Hadamard coefficients simultaneously, which reduces the influence of noise and dramatically speeds up data acquisition. We demonstrate that the SESPS with 32 spectral channels can accelerate spectral measurements from white light sources and fluorescence particles by around 14 times and 70 times, respectively, compared to measurements using a commercial spectrometer when the relative root mean square error (RMSE) is around 3% or smaller. The acceleration factors can be boosted by an extra 4 times when only eight spectral channels are used to achieve a compression ratio of 4:1, in which the relative RMSEs change only marginally. Compared to our previous SPS-HT, this new scheme can increase the speed by three orders of magnitude. This technique is expected to be useful in applications requiring high-speed spectral measurements such as the spectral flow cytometry and on-site medical diagnosis using fluorescence or Raman spectroscopy.
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