Measuring the primary cilium length: improved method for unbiased high-throughput analysis

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• 作者：Anneloes Dummer ; Christian Poelma ; Marco C. DeRuiter ; Marie-José T. H. Goumans…
• 关键词：Primary cilia ; Cilium length ; Measurements ; Methods
• 刊名：Cilia
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：5
• 期：1
• 全文大小：1,767 KB
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• 作者单位：Anneloes Dummer (1)
  Christian Poelma (2)
  Marco C. DeRuiter (1)
  Marie-José T. H. Goumans (3)
  Beerend P. Hierck (1)
  
  1. Department Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands
  2. Laboratory for Aero & Hydrodynamics, Delft University of Technology, Delft, The Netherlands
  3. Department Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands
  
• 刊物主题：Cell Biology; Developmental Biology; Human Genetics; Molecular Medicine; Biochemistry, general; Receptors;
• 出版者：BioMed Central
• ISSN：2046-2530

文摘

Background Primary cilia are cellular protrusions involved in mechanic and chemical sensing on almost all cells of our body. Important signaling pathways, including Hedgehog, TGFβ, and Ca2+, are linked to cilia and/or cilia function. Cilia can vary in length, which has functional implications. To measure these lengths correctly, a standardized method with high reliability and throughput is required. To date, methods for length measurements in cultured cells after fluorescent staining for ciliary components are error prone with a possible human selection bias, primarily caused by the orientation of cilia with respect of the imaging plane. In tissue sections, accurate measurements become an even larger challenge due to additional random sectioning plane. Cilia can be reconstructed in 3D and measured one by one, but this is a labor-intensive procedure. Therefore, we developed a new, high-throughput method with less selection bias.