Advanced nanopipette robotics for multiparametric imaging and manipulation of living cells

In this seminar, Dr Andrew Shevchuk will guide us through the past decade of advancements in Scanning Ion Conductance Microscopy (SICM) instrumentation and methodologies that transformed this glass nanopipette scanning technique. lnitially used to generate topographical images of living cells in physiological buffer mediums, it has soon evolved into a multiparametric live imaging tool with active manipulation capabilities. By combining SICM with fiuorescence confocal microscopy and, more recently, fiuorescence light sheet imaging, it is possible to link dynamic volumetric changes in cell 30 nanomorphology with fiuorescence signals from speciflc molecules involved. Furthermore, glass na­ nopipettes can be chemically modifled to convert them into electrochemical nanoprobes far localised measurement of pH, oxygen, ROS, and ATP, many of which can be performed in a correlative imaging manner. Despite being a truly non-invasive imaging technique capable of, but not limited to, 24-hour live cell topographical imaging, a methodology has been established to utilise SICM pipettes to apply forces in the hundreds of piconewtons range far low-stress mapping of cell mechanical properties with nanoscale resolution. SICM nanopipettes can also be employed far multiple rounds of controlled deli­ very and extraction of charged and neutral molecules via localised electroporation into and from the cell cytoplasm and nucleus, with minimal damage, as well as far detecting DNA, RNA, and proteins through translocation. Coupled with the ability far highly localised drug delivery via nanopipette, dose-response measurements, and electrophysiological recordings, ali these features combine to forma versatile, robo­ tic, multitool system suitable far a wide range of experiments aimed at disclosing subtle aspects of cell (patho)physiology and/or cell biophysics.

Registration email: angela_losada@ehu.eus