Abstract
Organ-on-a-chip technology is a 3D cell culture breakthrough of the last decade. This rapidly developing field of bioengineering intertwined with microfluidics provides new insights into disease development and preclinical drug screening. So far, optical and fluorescence microscopy are the most widely used methods to monitor and extract information from these models. Meanwhile transmission electron microscopy (TEM), despite its wide use for the characterization of nanomaterials and biological samples, remains unexplored in this area. In our work we propose a TEM sample preparation method, that allows to process a microfluidic chip without its prior deconstruction, into TEM-compatible specimens. We demonstrated preparation of tumor blood vessel-on-a-chip model and consecutive steps to preserve the endothelial cells lining microfluidic channel, for the chip’s further transformation into ultrathin sections. This approach allowed us to obtain cross-sections of the microchannel with cells cultured inside, and to observe cell adaptation to the channel geometry, as well as the characteristic for endothelial cells tight junctions. The proposed sample preparation method facilitates the electron microscopy ultrastructural characterization of biological samples cultured in organ-on-a-chip device.
Highlights
IntroductionIn the last 20 years we have witnessed an expansion of 3D cell culture models in the form of organ-on-a-chip (OoC) platforms that have revolutionized in vitro studies [1,2]
In this work we present the preparation of a PDMS-based 3D cell culture chip, suitable for post-processing and imaging with transmission electron microscopy
We have developed a method for microfluidic organ-on-a-chip cross section sample preparation compatible with transmission electron microscopy imaging to reveal ultrastructural cell characteristics
Summary
In the last 20 years we have witnessed an expansion of 3D cell culture models in the form of organ-on-a-chip (OoC) platforms that have revolutionized in vitro studies [1,2]. OoCs are an alternative tool in preclinical research to address the challenges in the development of new medicines, arising as a response to limited availability of human models, especially in the area of disease target identification, drug efficacy and toxicity studies [3,4]. OoCs are devices with one or more biocompatible microfluidic chambers, allowing the growth and maintenance of 3D cell culture under sterile and controlled conditions. The flexibility in design and operations makes microfluidic chips suitable to mimic broad range of physiological conditions in healthy organs or to induce disease pathology at a tissue level [9,10,11,12]. An important focus is placed on cancer vasculature, where its abnormal growth and morphology leads to enhanced permeability, that currently is the fundamental phenomenon used in design of drug nanocarriers to ensure their extravasation [14,15,16,17]
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