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White, B. P.

Publications and source records attributed to White, B. P..

4 recordsLinked to original sources

IN4MER Bioink: A Phosphorescent Biosensing Bio-ink for Multiple Analytes (Glucose, Lactate, Oxygen) Measurements and Temperature Sensing Applications

3D bioprinting has revolutionized tissue engineering by enabling researchers to create much more complex structures than was practical with earlier techniques. Bioprinting uses computer-controlled layer-by-layer deposition of a mixture of hydrogels and living cells and the resulting structures can mimic the complex geometries of many living tissues by incorporating multiple bioinks with varied material properties and cell populations, allowing researchers to design structures that vary not only in shape, but also in mechanical, chemical, and biological properties throughout the bioprinted construct. However, techniques for evaluating these living constructs and monitoring them over time have not yet caught up to these innovations. Here we describe a novel approach to reporting nutrient values in real-time throughout the scaffold itself, accomplished by dispersing oxygen, glucose, and lactate sensitive microspheres within bioinks. These can be noninvasively interrogated using low-cost phosphorescence lifetime readers to determine and track nutrient concentrations across our bioprinted constructs in real time. The wealth of information this technique produces suggests this may provide a powerful new tool for evaluating and designing future bioprinted constructs.

bioengineering↗

Polarization light-sheet microscopy and tomography (PLμTo) for flow-based imaging of 3D microcarrier mesenchymal stem cell culture

Light-sheet microscopy is an increasingly popular imaging tool for studying biological processes, however, it generally requires the use of exogenous fluorescent contrast agents which can be toxic to live cells. Light-sheet tomography is the label-free analog of light-sheet microscopy, utilizing elastic scattering contrast to visualize the structure of the biological sample. Here, we present a multi-modal polarization light-sheet microscopy and tomography (PL{micro}To) system for on-line, non-destructive, and non-invasive volumetric imaging of human mesenchymal stem cells, also referred to as mesenchymal stromal cells, attached to spherical gelatin methacryloyl microcarriers for image analysis-based quantitative monitoring of adherent stem cell culture expansion. The PL{micro}To system, which is built on an inverted selective plane illumination configuration, is compatible with standard horizontal sample mounting and microfluidics for multimodal volumetric data acquisition. This work is the first demonstration of scattering-based volume optical imaging flow cytometry.

biophysics↗

Comparison of polystyrene and hydrogel microcarriers for optical imaging of adherent cells

The ability to observe and monitor cell density and morphology has been imperative for assessing the health of a cell culture and for producing high quality, high yield cell cultures for decades. Microcarrier-based cultures, used for large-scale cellular expansion processes, are not compatible with traditional visualization-based methods such as widefield microscopy due to their thickness and material composition. Light-sheet tomography using label-free elastic scattering contrast from planar side illumination can achieve optical sectioning of thick samples, permitting non-invasive and non-destructive, in toto, three-dimensional, high-resolution visualization of cells cultured on microcarriers. Here, we assess the optical imaging capabilities of commercial polystyrene microcarriers versus custom-fabricated gelatin methacrylate (gelMA) microcarriers for non-destructive and non-invasive visualization of the entire microcarrier surface, direct cell enumeration, and sub-cellular visualization of mesenchymal stem/stromal cells (MSCs). The polystyrene microcarrier prevents visualization of cells on the distal half of the microcarrier using either fluorescence or elastic scattering contrast, whereas the gelMA microcarrier allows high fidelity visualization of cell morphology and quantification of cell density using light sheet fluorescence microscopy and tomography. The combination of optical-quality gelMA microcarriers and label-free light-sheet tomography will facilitate enhanced control of bioreactor-microcarrier cell culture processes.

biophysics↗

Volumetric imaging of human mesenchymal stem cells (hMSCs) for non-destructive quantification of 3D cell culture growth

The adoption of cell-based therapies into the clinic will require tremendous large-scale expansion to satisfy future demand, and bioreactor-microcarrier cultures are best suited to meet this challenge. The use of spherical microcarriers, however, precludes in-process visualization and monitoring of cell number, morphology, and culture health. The development of novel expansion methods also motivates the advancement of analytical methods used to characterize these microcarrier cultures. A robust optical imaging and image-analysis assay to non-destructively quantify cell number and cell volume was developed. This method preserves 3D cell morphology and does not require membrane lysing, cellular detachment, or exogenous labeling. Complex cellular networks formed in microcarrier aggregates were imaged and analyzed in toto. Direct cell enumeration of large aggregates was performed in toto for the first time. This assay was successfully applied to monitor cellular growth of mesenchymal stem cells attached to spherical hydrogel microcarriers over time. Elastic scattering and fluorescence lightsheet microscopy were used to quantify cell volume and cell number at varying spatial scales. The presented study motivates the development of on-line optical imaging and image analysis systems for robust, automated, and non-destructive monitoring of bioreactor-microcarrier cell cultures.

biophysics↗