title: Cellular and Nuclear Alignment Analysis for Determining Epithelial Cell Chirality.
abstract: Left-right (LR) asymmetry is a biologically conserved property in living organisms that can be observed in the asymmetrical arrangement of organs and tissues and in tissue morphogenesis, such as the directional looping of the gastrointestinal tract and heart. The expression of LR asymmetry in embryonic tissues can be appreciated in biased cell alignment. Previously an in vitro chirality assay was reported by patterning multiple cells on microscale defined geometries and quantified the cell phenotype-dependent LR asymmetry, or cell chirality. However, morphology and chirality of individual cells on micropatterned surfaces has not been well characterized. Here, a Python-based algorithm was developed to identify and quantify immunofluorescence stained individual epithelial cells on multicellular patterns. This approach not only produces results similar to the image intensity gradient-based method reported previously, but also can capture properties of single cells such as area and aspect ratio. We also found that cell nuclei exhibited biased alignment. Around 35% cells were misaligned and were typically smaller and less elongated. This new imaging analysis approach is an effective tool for measuring single cell chirality inside multicellular structures and can potentially help unveil biophysical mechanisms underlying cellular chiral bias both in vitro and in vivo.
authors: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Biotech 2147, 110 8th Street, Troy, NY, 12180, USA.
authors: Michael J
authors: MJ
authors: Raymond
authors: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Biotech 2147, 110 8th Street, Troy, NY, 12180, USA.|Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA.
authors: Poulomi
authors: P
authors: Ray
authors: Department of Biology, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA.
authors: Gurleen
authors: G
authors: Kaur
authors: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Biotech 2147, 110 8th Street, Troy, NY, 12180, USA.|Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA.|Department of Physical Intelligence, Max Planck Institute for Intelligent Systems, Heisenbergstr 3, 70569, Stuttgart, Germany.
authors: Ajay V
authors: AV
authors: Singh
authors: Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Biotech 2147, 110 8th Street, Troy, NY, 12180, USA. wanq@rpi.edu.|Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA. wanq@rpi.edu.|Center for Modeling, Simulation and Imaging in Medicine, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA. wanq@rpi.edu.
authors: Leo Q
authors: LQ
authors: Wan
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D016764:Cell Polarity / Q000502:physiology*; D004285:Dogs; D061985:Madin Darby Canine Kidney Cells
keywords: Cell alignment; Cell morphology; Intensity gradient; Micropatterning; Nucleus alignment