Chronic label-free volumetric photoacoustic microscopy of melanoma cells in three-dimensional porous Scaffolds

Abstract

Visualizing cells in three-dimensional (3D) scaffolds has been one of the major challenges in tissue engineering. Most current imaging modalities either suffer from poor penetration depth or require exogenous contrast agents. Here, we demonstrate photoacoustic microscopy (PAM) of the spatial distribution and temporal proliferation of cells inside three-dimensional porous scaffolds with thicknesses over 1 mm. Specifically, we evaluated the effects of seeding and culture methods on the spatial distribution of melanoma cells. Spatial distribution of the cells in the scaffold was wellresolved in PAM images. Moreover, the number of cells in the scaffold was quantitatively measured from the as-obtained volumetric information. The cell proliferation profile obtained from PAM correlated well with what was obtained using the traditional 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

Imaging Strategies for Tissue Engineering Applications

“…8A). [112][113][114] In vivo animal studies using multiple PA imaging systems have been demonstrated, including a single element transducer-based microscopy, linear array-based imaging, and tomography. With its high spatial resolution and sensitivity, PA microscopy was useful to visualize vascularization and scaffold degradation in the nude mouse ear model as shown in Figure 8B.…”

“…Reprinted with permission from Zhang et al 112 (B) Optical-resolution photoacoustic microscopy (top), OCT (middle), and combined (bottom) images of neovascularization within inverse opal scaffolds implanted in a mouse ear model. Reprinted with permission from Cai et al 115 (C) Combined ultrasound and spectroscopic photoacoustic imaging of gold nanoparticle-labeled mesenchymal stem cells intramuscularly injected within a PEGylated fibrin hydrogel.…”

The Laser Technology: New Trends in Biology and Medicine

“…Photo-acoustic imaging uses the intrinsic chromophore, which has an optical absorption signature, as long as appropriate irradiation wavelengths are applied. It is used for example for haemoglobin [66], melanin [67], water [68] or lipid [69].…”

Penerapan Sistem Kendali XY-Stage dan Modulasi Laser Pada Tomografi Fotoakustik Menggunakan Arduino

“…Kombinasi sistem pencitraan fotoakustik dengan melibatkan pencitraan optik dan ultrasonic mampu menghasilkan gambar resolusi tinggi dengan kedalaman penetrasi yang baik [5]. Visualisasi sel dalam tiga dimensi (3D) telah menjadi salah satu tantangan utama dalam rekayasa jaringan, telah diterapkan mikroskopi fotoakustik dari distribusi spasial dan proliferasi temporal sel dalam perancah porous tiga dimensi dengan ketebalan lebih dari 1 mm [6].…”