Biophysical Society 62nd Annual Meeting | Program Guide

2:30 PM–4:00 PM Allen Institute for Cell Science The Allen Institute for Cell Science – Resources to Empower Your Research The Allen Institute for Cell Science aims to understand and predict behavior of human cells in health and disease. We have chosen the induced pluripotent human stem cell as our model because it is diploid, proliferative, and differentiates in a number of different cell types. In this presentation, the Allen Institute for Cell Science team will intro- duce you to the publicly available cell lines, observations, imaging and computational methods and tools, and the data produced by the Insti- tute. We will discuss our legacy collection of endogenous fluorescently tagged human induced pluripotent stem cell lines highlighting key in- tracellular structures, and how we image our cells in our high-replicate microscopy pipeline, that includes automated cell culture and imaging using spinning disk microscopy. We will also discuss our workflow quality control criteria, the methods developed to ensure day-to-day consistency between data sets, and how alternate pipeline modes may offer the flexibility to evaluate new assays and imaging technologies. We have collected 3D, 4 channel images frommore than 20,000 live cells thus far, comprised of high replicates for each genome-edited cell line. This data offers ideal input for key analyses examining variation in the cell population and machine learning. We will demonstrate this using some easily accessible tools for descriptive statistical analyses de- veloped in-house. We will also show how this rich, high-replicate image set is used as input for deep neural networks which generate unified, integrated cell models and label free imaging. Finally, we’ll demonstrate how to navigate our large, high replicate 3D image data sets, revealing the subcellular localization of key tagged structures. All of our procedures, tools, and data are shared on our webpage, the Allen Cell Explorer (www.allencell.org), which will be highlighted during the presentation. Speakers Allen Institute for Cell Science team

12:30 PM–2:00 PM Alvéole Maskless Quantitative Multi-Protein Photopatterning to Orchestrate Cellular Microenvironment Cell biology is faced with significant challenges when attempting to create complex microenvironments to unravel intricate mechanisms in- volved in cell adhesion, cell polarity, cell migration, etc. These challeng- es can be overcome by molecular printing which involves the controlled deposition of molecules on a substrate at the micrometer scale. These approaches have developed tremendously in the past few years and mi- cropatterned substrates are now routinely used for biological research. To yield biologically relevant data, printed biomolecules should mimic the complexity of the in vivo microenvironment. Micrometer-scale gradients of multiple proteins are thus highly desirable. Here we present PRIMO custommicropatterning system for cell control which allows to control the chemistry and topography of the cellular microenvironment and study their impacts on cell development. This maskless quantitative multi-protein photopatterning solution is based on the light-induced molecular adsorption of proteins (LIMAP) technology. The PRIMO system combines a UV illumination module and a specific photoactivatable reagent (PLPP). The combined action of UV- light and PLPP locally degrades antifouling polymer brushes allowing for the adsorption of proteins in a well-defined area. PRIMO relies on a wide-field DMD-based projection system coupled to an epifluorescence microscope to project custom-defined patterns of UV light onto all standard cell culture surface. As a result, micrometer scale patterns are generated within seconds. The remaining background allows for the sequential patterning of multiple proteins. Controlled protein gradients of custom-defined shape can also be patterned. In addition, PRIMO technology allows for microfabrication by photopoly- merization of UV-sensitive materials and also protein patterning onto pre-existing 3D surfaces. This new micropatterning technology empowers biomedical research in neurobiology, immunology, stem cell biology, oncology, and tissue engineering. The applications in cell biology, such as studying how the asymmetry of the focal adhesion can regulate the cytoskeleton, will be illustrated by some user testimonials presenting their research works conducted with PRIMO. Visit www.alveolelab.com for more information. Speaker Pierre-Olivier Strale, Senior Scientist, Alvéole

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