Combined ion conductance and fluorescence confocal microscopy for biological cell membrane transport studies

Optical visualization of nanoscale morphological changes taking place in living biological cells during such important processes as endo- and exocytosis is challenging due to the low refractive index of lipid membranes. In this paper we summarize and discuss advances in the powerful combination of two complementary live imaging techniques, ion conductance and fluorescence confocal microscopy, that allows cell membrane topography to be related with molecular-specific fluorescence at high spatial and temporal resolution. We demonstrate the feasibility of the use of ion conductance microscopy to image apical plasma membrane of mouse embryo trophoblast outgrowth cells at a resolution sufficient to depict single endocytic pits. This opens the possibility to study individual endocytic events in embryo trophoblast outgrowth cells where endocytosis plays a crucial role during early stages of embryo development.

[1]  D. Perrais,et al.  A High Precision Survey of the Molecular Dynamics of Mammalian Clathrin-Mediated Endocytosis , 2011, Microscopy and Microanalysis.

[2]  Y. Korchev,et al.  Specialized scanning ion‐conductance microscope for imaging of living cells , 1997, Journal of microscopy.

[3]  Liming Ying,et al.  Programmable delivery of DNA through a nanopipet. , 2002, Analytical chemistry.

[4]  P. Hansma,et al.  The scanning ion-conductance microscope. , 1989, Science.

[5]  H. McMahon,et al.  Mechanisms of endocytosis. , 2009, Annual review of biochemistry.

[6]  M J Lab,et al.  Scanning ion conductance microscopy of living cells. , 1997, Biophysical journal.

[7]  J. Gorelik,et al.  Comparison of the arrhythmogenic effects of tauro‐ and glycoconjugates of cholic acid in an in vitro study of rat cardiomyocytes , 2004, BJOG : an international journal of obstetrics and gynaecology.

[8]  Achim Hartschuh,et al.  Tip-enhanced near-field optical microscopy. , 2008, Angewandte Chemie.

[9]  S. Hell,et al.  Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. , 1994, Optics letters.

[10]  Sara A Love,et al.  Assessing nanoparticle toxicity. , 2012, Annual review of analytical chemistry.

[11]  D. Klenerman,et al.  Simultaneous measurement of Ca2+ and cellular dynamics: combined scanning ion conductance and optical microscopy to study contracting cardiac myocytes. , 2001, Biophysical journal.

[12]  J. Gorelik,et al.  Dexamethasone and ursodeoxycholic acid protect against the arrhythmogenic effect of taurocholate in an in vitro study of rat cardiomyocytes , 2003, BJOG : an international journal of obstetrics and gynaecology.

[13]  Richard W. Clarke,et al.  Realizing the biological and biomedical potential of nanoscale imaging using a pipette probe. , 2011, Nanomedicine.

[14]  Y. Korchev,et al.  Taurocholate induces changes in rat cardiomyocyte contraction and calcium dynamics. , 2002, Clinical science.

[15]  Thomas D. Schmittgen,et al.  Dynamin 2 Regulates Riboflavin Endocytosis in Human Placental Trophoblasts , 2007, Molecular Pharmacology.

[16]  David Klenerman,et al.  Imaging proteins in membranes of living cells by high-resolution scanning ion conductance microscopy. , 2006, Angewandte Chemie.

[17]  I. Sargent,et al.  Caveolin-1 and lipid rafts in confluent BeWo trophoblasts: evidence for Rock-1 association with caveolin-1. , 2007, Placenta.

[18]  T. Harris,et al.  Leucine and arginine regulate trophoblast motility through mTOR-dependent and independent pathways in the preimplantation mouse embryo. , 2012, Developmental biology.

[19]  D. Klenerman,et al.  Scanning surface confocal microscopy for simultaneous topographical and fluorescence imaging: Application to single virus-like particle entry into a cell , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Kazuhiko Ishihara,et al.  Cell membrane-inspired phospholipid polymers for developing medical devices with excellent biointerfaces , 2012, Science and technology of advanced materials.

[21]  T. Schäffer,et al.  Comparison of scanning ion conductance microscopy with atomic force microscopy for cell imaging. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[22]  V. Perry,et al.  Adaptive Responses by Mouse Early Embryos to Maternal Diet Protect Fetal Growth but Predispose to Adult Onset Disease1 , 2008, Biology of reproduction.

[23]  E. Eisenberg,et al.  Clathrin exchange during clathrin-mediated endocytosis , 2001, The Journal of cell biology.

[24]  Yan Zhang,et al.  Real-Time Investigation of Engineered Nanomaterials Cytotoxicity in Living Alveolar Epithelia with Hopping Probe Ion Conductance Microscopy , 2013 .

[25]  J. Simpson,et al.  Impact of live cell imaging on coated vesicle research. , 2007, Seminars in cell & developmental biology.

[26]  T. Schäffer,et al.  Image formation, resolution, and height measurement in scanning ion conductance microscopy , 2009 .

[27]  Y. Korchev,et al.  Immortalization of human alveolar epithelial cells to investigate nanoparticle uptake. , 2008, American journal of respiratory cell and molecular biology.

[28]  C. Lamaze,et al.  Stressing caveolae new role in cell mechanics. , 2012, Trends in cell biology.

[29]  X. Zhuang,et al.  Fast three-dimensional super-resolution imaging of live cells , 2011, Nature Methods.

[30]  John A.G. Briggs,et al.  Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision , 2011, The Journal of cell biology.

[31]  M. Tremblay,et al.  A clathrin, caveolae, and dynamin-independent endocytic pathway requiring free membrane cholesterol drives HIV-1 internalization and infection in polarized trophoblastic cells. , 2007, Journal of molecular biology.

[32]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[33]  D. Klenerman,et al.  Nanoscale live-cell imaging using hopping probe ion conductance microscopy , 2009, Nature Methods.

[34]  M J Lab,et al.  Cell volume measurement using scanning ion conductance microscopy. , 2000, Biophysical journal.

[35]  D. Klenerman,et al.  Imaging single virus particles on the surface of cell membranes by high-resolution scanning surface confocal microscopy. , 2008, Biophysical journal.

[36]  Y. Korchev,et al.  Esmolol is antiarrhythmic in doxorubicin‐induced arrhythmia in cultured cardiomyocytes – determination by novel rapid cardiomyocyte assay , 2003, FEBS letters.

[37]  D. Klenerman,et al.  An alternative mechanism of clathrin-coated pit closure revealed by ion conductance microscopy , 2012, The Journal of cell biology.

[38]  Ari Helenius,et al.  Virus entry by endocytosis. , 2010, Annual review of biochemistry.

[39]  I. Cameron,et al.  Metabolic Induction and Early Responses of Mouse Blastocyst Developmental Programming following Maternal Low Protein Diet Affecting Life-Long Health , 2012, PloS one.

[40]  Alan Morgan,et al.  Secretory granule exocytosis. , 2003, Physiological reviews.

[41]  D. Klenerman,et al.  Endocytic pathways: combined scanning ion conductance and surface confocal microscopy study , 2008, Pflügers Archiv - European Journal of Physiology.

[42]  I. Dietzel,et al.  Monitoring cell movements and volume changes with pulse‐mode scanning ion conductance microscopy , 2003, Journal of microscopy.

[43]  Elina Ikonen,et al.  When intracellular logistics fails - genetic defects in membrane trafficking , 2006, Journal of Cell Science.

[44]  S. Simon,et al.  Imaging with total internal reflection fluorescence microscopy for the cell biologist , 2010, Journal of Cell Science.

[45]  Julia Gorelik,et al.  Dynamic assembly of surface structures in living cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[46]  C. Xie,et al.  Tip-enhanced fluorescence microscopy of high-density samples , 2006 .