University Lyon 1

7 articles published in JoVE

• Cancer Research

  
  Semi-automatic PD-L1 Characterization and Enumeration of Circulating Tumor Cells from Non-small Cell Lung Cancer Patients by Immunofluorescence Jessica Garcia^1,2,3, David Barthelemy^1,2,3, Florence Geiguer^1,2, Julie Ballandier^1,2,3, Kathryn W. Li⁴, Jean-Phillippe Aurel⁴, Frédérique Le Breton⁵, Claire Rodriguez-Lafrasse¹, Brigitte Manship³, Sébastien Couraud^6,7, Léa Payen^{1,2,3 1}Laboratoire de Biochimie et Biologie Moléculaire, Groupe Hospitalier Sud, Hospices Civils de Lyon, ²Circulating Cancer (CIRCAN) Program, Hospices Civils de Lyon Cancer Institute, ³University of Lyon, Claude Bernard University, Cancer Research Center of Lyon, ⁴Biolidics Limited, ⁵Institut de Pathologie Multisites des HCL-Site Sud, Hospices Civils de Lyon, ⁶Acute Respiratory Disease and Thoracic Oncology Department, Lyon Sud Hospital, Hospices Civils de Lyon Cancer Institute, ⁷EMR-3738 Therapeutic Targeting in Oncology, Lyon Sud Medical Faculty, Lyon 1 University The characterization of circulating tumor cells (CTCs) is a popular topic in translational research. This protocol describes a semi-automatic immunofluorescence (IF) assay for PD-L1 characterization and enumeration of CTCs in non-small cell lung cancer (NSCLC) patient samples.

• Chemistry

  
  Analysis of Minerals Produced by hFOB 1.19 and Saos-2 Cells Using Transmission Electron Microscopy with Energy Dispersive X-ray Microanalysis Lukasz Bozycki¹, Magdalena Komiazyk¹, Saida Mebarek^2,3,4,5,6, Rene Buchet^2,3,4,5,6, Slawomir Pikula¹, Agnieszka Strzelecka-Kiliszek^{1 1}Laboratory of Lipid Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, ²Université de Lyon, ³Université Lyon 1, ⁴ We present a protocol to compare the state of minerals in vesicles released by two human bone cell lines: hFOB 1.19 and Saos-2. Their mineralization profiles were analyzed by Alizarin Red-S (AR-S) staining, ultraviolet (UV) light visualization, transmission electron microscopy (TEM) imaging and energy dispersive X-ray microanalysis (EDX).

• Biology

  
  Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells Emily Tubbs¹, Jennifer Rieusset^{2 1}Lund University Diabetes Centre, Lund University, ²INSERM UMR-1060, CarMeN Laboratory, Lyon 1 University, INRA U1235, INSA of Lyon, Rockefeller and Charles Merieux Lyon-Sud Medical Universities Here, we describe a procedure to visualize and quantify with high sensitivity the endogenous interactions between the endoplasmic reticulum and mitochondria in fixed cells. The protocol features an optimized in situ proximity ligation assay targeting the inositol 1,4,5-triphosphate receptor/glucose-regulated protein 75/voltage-dependent anion channel/cyclophilin D complex at the mitochondria-associated membrane interface.

• Medicine

  
  Confocal Time Lapse Imaging as an Efficient Method for the Cytocompatibility Evaluation of Dental Composites Ghania Nina Attik¹, Kerstin Gritsch^1,2, Pierre Colon^1,3, Brigitte Grosgogeat^{1,2 1}Laboratoire des Multimatériaux et Interfaces, UMR CNRS 5615, Université Lyon1, ²UFR d'Odontologie, Université Lyon1; Service de Consultations et de Traitements Dentaires, Hospices Civils de Lyon, ³UFR d'Odontologie, Université Paris Diderot; Service d'Odontologie, APHP, Hôpital Rothschild The most studied aspect of the biocompatibility of dental composites is their cytotoxicity 3D CLSM time lapse imaging combined with fluorescent signal quantification allows sensitive evaluation of the cell fate over time. Utilizing this method could be an efficient tool to assess the cytocompatibility of dental composites.

• Neuroscience

  
  Culture of Isolated Floor Plate Tissue and Production of Conditioned Medium to Assess Functional Properties of Floor Plate-released Signals Camille Charoy¹, Elise Arbeille¹, Karine Thoinet¹, Valérie Castellani^{1 1}CGphiMC, UMR CNRS 5534, University of Lyon 1 The floor plate is a crucial structure of the developing spinal cord, providing diffusible signals to progenitors and differentiating neurons. We describe a method to produce floor-plate conditioned medium, to apply it to fresh spinal cord tissue, and to assess the consequences on proteins of interest by biochemistry.

• Neuroscience

  
  Detection of the Genome and Transcripts of a Persistent DNA Virus in Neuronal Tissues by Fluorescent In situ Hybridization Combined with Immunostaining Frédéric Catez^1,2,5, Antoine Rousseau⁴, Marc Labetoulle⁴, Patrick Lomonte^{1,2,3 1}Virus and Centromere Team, Centre de Génétique et Physiologie Moléculaire et Cellulaire, CNRS UMR 5534, ²Université de Lyon 1, ³Laboratoire d'excellence, LabEX DEVweCAN, ⁴Institut de Virologie Moléculaire et Structurale, CNRS UPR 3296, ⁵Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS UMR 5286 We established a fluorescent in situ hybridization protocol for the detection of a persistent DNA virus genome within tissue sections of animal models. This protocol enables studying infection process by codetection of the viral genome, its RNA products, and viral or cellular proteins within single cells.

• Medicine

  
  Microarray-based Identification of Individual HERV Loci Expression: Application to Biomarker Discovery in Prostate Cancer Philippe Pérot^1,2, Valérie Cheynet^1,2, Myriam Decaussin-Petrucci^1,3,4, Guy Oriol^1,2, Nathalie Mugnier⁵, Claire Rodriguez-Lafrasse^1,4,6, Alain Ruffion^1,4,7, François Mallet^{1,2 1}Joint Unit Hospices de Lyon-bioMérieux, ²Medical Diagnostic Discovery Department, BioMérieux, ³Department of Pathology and Cytology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, ⁴Medical Faculty, Lyon 1 University, ⁵Data and Knowledge Laboratory, BioMérieux, ⁶Department of Biochemistry and Molecular Biology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, ⁷Department of Urology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon Human endogenous retroviruses (HERV), which occupy 8% of the human genome, retain scarce coding capacities but a hundred thousand long terminal repeats (LTRs). A custom Affymetrix microarray was designed to identify individual HERV locus expression and was used on prostate cancer tissues as a proof of concept for future clinical studies.