Effects of substrate patterning on cellular spheroid growth and dynamics measured by gradient light interference microscopy (GLIM)

Abstract

The development of three‐dimensional (3D) cellular architectures during development and pathological processes involves intricate migratory patterns that are modulated by genetics and the surrounding microenvironment. The substrate composition of cell cultures has been demonstrated to influence growth, proliferation and migration in 2D. Here, we study the growth and dynamics of mouse embryonic fibroblast cultures patterned in a tissue sheet which then exhibits 3D growth. Using gradient light interference microscopy (GLIM), a label‐free quantitative phase imaging approach, we explored the influence of geometry on cell growth patterns and rotational dynamics. We apply, for the first time to our knowledge, dispersion‐relation phase spectroscopy (DPS) in polar coordinates to generate the radial and rotational cell mass‐transport. Our data show that cells cultured on engineered substrates undergo rotational transport in a radially independent manner and exhibit faster vertical growth than the control, unpatterned cells. The use of GLIM and polar DPS provides a novel quantitative approach to studying the effects of spatially patterned substrates on cell motility and growth.

Multiscale Assay of Unlabeled Neurite Dynamics Using Phase Imaging with Computational Specificity

“…Our results highlight that mass transport and growth, two potentially independent phenomena, are anti-correlated in the neurites -where high growth rates appear accompanied by slower transport. This observation is in-line with the understanding that there exists a metabolic trade-off between cellular motion and vegetative growth [65][66][67][68] The complexity of imaging neuronal clusters is primarily due to the photochemical sensitivity of the cells and the resolving power needed to detect fine structures such as dendrites within a developing arbor. To meet these challenges, our approach relies on machine learning, quantitative phase imaging, and transport analysis with few conventional analogs.…”

“…This hypothesis is further supported by the interplay of intracellular cellular transport and neurite growth. By limiting our imaging to slow time scales, we focused primarily on anteretrograde transport motion 65 . In Fig.…”

3D printing of tissue engineering scaffolds: a focus on vascular regeneration

“…Usually, there are three methods for the formation, including microfluidics, spheroids on matrices, and hanging drop techniques [ 57 ]. Cell spheroids can be directly formed on patterned matrices [ 58 ]. In microfluidic methods, cell spheroids are formed by fabricating channels with different structures to manipulate fluids [ 59 ].…”

Computational interference microscopy enabled by deep learning

“…[2] As the specimen refractive index reports on the dry mass density, QPI has been employed to studying cell growth [3][4][5][6][7]. Analyzing the spatio-temporal fluctuations of dry mass provided a new way of monitoring intracellular transport and differentiating between diffusive and active processes [8][9][10]. Due to its sensitivity to nanometer scale optical pathlength changes, QPI is capable of measuring cell membrane fluctuations [11][12][13][14] and imaging unlabeled single microtubules [15].…”