Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

Abstract

The past decade has seen an increasing demand for more complex, reproducible and physiologically relevant tissue cultures that can mimic the structural and biological features of living tissues. Monitoring the viability, development and responses of such tissues in real‐time are challenging due to the complexities of cell culture physical characteristics and the environments in which these cultures need to be maintained in. Significant developments in optics, such as optical manipulation, improved detection and data analysis, have made optical imaging a preferred choice for many three‐dimensional (3D) cell culture monitoring applications. The aim of this review is to discuss the challenges associated with imaging and monitoring 3D tissues and cell culture, and highlight topical label‐free imaging tools that enable bioengineers and biophysicists to non‐invasively characterise engineered living tissues.

Review of optical methods for noninvasive imaging of skin fibroblasts—From in vitro to ex vivo and in vivo visualization

“…At the moment, there are publications about using the SRS for studying 3D cultures. Potentially, by using SRS and CARS microscopy to study 3D fibroblast cultures, researchers can gain insight into how fibroblasts interact and modify the ECM to assess tissue health based on metabolic balance [77]. In vivo applications can become the next step for this approach.…”

“…For that, fibroblasts were treated with cytochalasin D or colchicine to destroy actin filaments or microtubules, T A B L E 1 Optical techniques used for skin imaging (adapted from Refs. [49] and [77]). respectively.…”

Imagining the future of optical microscopy: everything, everywhere, all at once

“…Quantitative phase imaging 132 , 133 , stimulated Raman microscopy 134 – 136 (Fig. 2c ), as well as second harmonic generation and other orthogonal imaging techniques 25 – 28 can all be brought in to supplement and complement multiplexed fluorescence microscopy.…”

Advanced Online Monitoring of In Vitro Human 3D Full-Thickness Skin Equivalents

“…Despite the improvement of sophisticated tissue equivalents for the investigation of pathophysiological processes, the investigation of the respective 3D equivalents constitutes the current limitation to replace animal models and exploit the full potential of tissue equivalents. Especially for complex 3D tissue equivalents, it is crucial to develop minimally invasive non-destructive analytical methods to facilitate continuous or repetitive analysis and monitoring of tissue viability, development, functionality, and cellular responses reliably, reproducibly, and preferably in real time [ 20 , 21 ].…”