title: An integrated chip for immunofluorescence and its application to analyze lysosomal storage disorders.
abstract: Immunofluorescence (IF) is a common method to observe protein distribution and localization at the single-cell level through wide-field fluorescence or confocal microscopy. Conventional protocol for IF staining of cells typically requires a large amount of reagents, especially antibodies, and noticeable investment in both labor and time. Microfluidic technologies provide a cost-effective alternative: it can evaluate and optimize experimental conditions, and perform automatic and high-throughput IF staining on-chip. We employed this method to analyze lysosomal storage disorders (LSDs) based on the expression and morphological distribution of LAMP1 and LC3 in starving cells. With pneumatic valves integrated on-chip, the parallel staining process can be completed within a few hours. The total consumption of each antibody solution for the whole experiment is merely 0.3 μL. This device provides a promising tool for automated high-throughput molecular imaging at cell level that can be applied for diagnostic analysis.
authors: College of Engineering, and Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China.
authors: Jie
authors: J
authors: Shen
authors: Ying
authors: Y
authors: Zhou
authors: Tu
authors: T
authors: Lu
authors: Junya
authors: J
authors: Peng
authors: Zhixiang
authors: Z
authors: Lin
authors: Lei
authors: L
authors: Huang
authors: Yuhong
authors: Y
authors: Pang
authors: Li
authors: L
authors: Yu
authors: Yanyi
authors: Y
authors: Huang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D001343:Autophagy; D002460:Cell Line; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005455:Fluorescent Antibody Technique / Q000295:instrumentation*; D016464:Lysosomal Storage Diseases / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D052118:Lysosomal-Associated Membrane Protein 1 / Q000276:immunology / Q000378:metabolism; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000276:immunology / Q000378:metabolism; D051381:Rats
chemical_list: D000906:Antibodies; D016650:Fluorescein-5-isothiocyanate; D052118:Lysosomal-Associated Membrane Protein 1; D008869:Microtubule-Associated Proteins; C526109:Anxa3 protein, rat