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New tools for labeling silica in living diatoms
Authors:Desclés Julien  Vartanian Mathieu  El Harrak Abdeslam  Quinet Michelle  Bremond Nicolas  Sapriel Guillaume  Bibette Jérome  Lopez Pascal J
Institution:Laboratoire Biologie Moléculaire des Organismes Photosynthétiques CNRS UMR-8186, Ecole Normale Supérieure, 75005 Paris, France.
Abstract:Silicon biomineralization is a widespread mechanism found in several kingdoms that concerns both unicellular and multicellular organisms. As a result of genomic and molecular tools, diatoms have emerged as a good model for biomineralization studies and have provided most of the current knowledge on this process. However, the number of techniques available to study its dynamics at the cellular level is still rather limited. Here, new probes were developed specifically to label the pre-existing or the newly synthesized silica frustule of several diatoms species. It is shown that the LysoTracker Yellow HCK-123, which can be used to visualize silica frustules with common filter sets, presents an enhanced signal-to-noise ratio and allows details of the frustules to be imaged without of the use of ionophores. It is also demonstrated that methoxysilane derivatives can be coupled to fluorescein-5-isothiocyanate (FITC) to preferentially label the silica components of living cells. The coupling of labeling procedures might help to address the challenging question of the process of frustule exocytosis.
Keywords:3D-imaging  biomineralization  diatoms  exocytosis  nanopattern
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