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Scanning ion conductance microscopy: a convergent high-resolution technology for multi-parametric analysis of living cardiovascular cells.

Miragoli M, Moshkov A, Novak P, Shevchuk A, Nikolaev VO, El-Hamamsy I, Potter CM, Wright P, Kadir SH, Lyon AR, Mitchell JA, Chester AH, Klenerman D, Lab MJ, Korchev YE, Harding SE, Gorelik J - J R Soc Interface (2011)

Bottom Line: At the cellular level, heart failure leads to a pronounced loss of T-tubules in cardiac myocytes accompanied by a reduction in Z-groove ratio.The SICM pipette can be used for patch-clamp recordings of membrane potential and single channel currents.In conclusion, SICM provides a highly informative multimodal imaging platform for functional analysis of the mechanisms of cardiovascular diseases, which should facilitate identification of novel therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Science, National Heart and Lung Institute, Imperial College London, , Dovehouse Street, London SW36LY, UK.

ABSTRACT
Cardiovascular diseases are complex pathologies that include alterations of various cell functions at the levels of intact tissue, single cells and subcellular signalling compartments. Conventional techniques to study these processes are extremely divergent and rely on a combination of individual methods, which usually provide spatially and temporally limited information on single parameters of interest. This review describes scanning ion conductance microscopy (SICM) as a novel versatile technique capable of simultaneously reporting various structural and functional parameters at nanometre resolution in living cardiovascular cells at the level of the whole tissue, single cells and at the subcellular level, to investigate the mechanisms of cardiovascular disease. SICM is a multimodal imaging technology that allows concurrent and dynamic analysis of membrane morphology and various functional parameters (cell volume, membrane potentials, cellular contraction, single ion-channel currents and some parameters of intracellular signalling) in intact living cardiovascular cells and tissues with nanometre resolution at different levels of organization (tissue, cellular and subcellular levels). Using this technique, we showed that at the tissue level, cell orientation in the inner and outer aortic arch distinguishes atheroprone and atheroprotected regions. At the cellular level, heart failure leads to a pronounced loss of T-tubules in cardiac myocytes accompanied by a reduction in Z-groove ratio. We also demonstrated the capability of SICM to measure the entire cell volume as an index of cellular hypertrophy. This method can be further combined with fluorescence to simultaneously measure cardiomyocyte contraction and intracellular calcium transients or to map subcellular localization of membrane receptors coupled to cyclic adenosine monophosphate production. The SICM pipette can be used for patch-clamp recordings of membrane potential and single channel currents. In conclusion, SICM provides a highly informative multimodal imaging platform for functional analysis of the mechanisms of cardiovascular diseases, which should facilitate identification of novel therapeutic strategies.

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Schematic illustration of scanning ion conductance microscopy as a tool to study tissues and cells at the macroscopic, microscopic and nanoscopic levels of organization. (Online version in colour.)
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RSIF20100597F1: Schematic illustration of scanning ion conductance microscopy as a tool to study tissues and cells at the macroscopic, microscopic and nanoscopic levels of organization. (Online version in colour.)

Mentions: Scanning ion conductance microscopy (SICM) invented by Hansma et al. [13] lately has been developed to image and analyse surface topography of live cells in our group [14–16]. SICM is a non-optical method that uses a nanopipette as a scanning probe to image cell surface structures with nanometre resolution [16]. SICM and a battery of associated innovative methods are unique among current imaging techniques, not only in spatial resolution, but also in the rich combination of imaging modalities with other functional and dynamic methods [10,17,18]. Recently, we have developed a hopping probe ion conductance microscopy (HPICM) [15], using a concept of ‘hopping’ from one imaging point to another, first implemented in SICM as the pulse mode back-step SICM mode [19]. Unlike previous attempts based on this concept, the HPICM managed to obtain nanoscale resolution in highly convoluted live cell samples without compromising the scan speed [15], and has already led to more elaborate techniques for single particle tracking [20] and functional imaging of receptor distribution [18]. The aim of this review is to describe the SICM technique alone or in combination with other optical and electrical methods to perform highly resolved dynamic and integrative analysis of cardiac structure, physiology and mechanisms of cardiovascular disease at the subcellular, cellular and tissue levels (figure 1).Figure 1.


Scanning ion conductance microscopy: a convergent high-resolution technology for multi-parametric analysis of living cardiovascular cells.

Miragoli M, Moshkov A, Novak P, Shevchuk A, Nikolaev VO, El-Hamamsy I, Potter CM, Wright P, Kadir SH, Lyon AR, Mitchell JA, Chester AH, Klenerman D, Lab MJ, Korchev YE, Harding SE, Gorelik J - J R Soc Interface (2011)

Schematic illustration of scanning ion conductance microscopy as a tool to study tissues and cells at the macroscopic, microscopic and nanoscopic levels of organization. (Online version in colour.)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3104336&req=5

RSIF20100597F1: Schematic illustration of scanning ion conductance microscopy as a tool to study tissues and cells at the macroscopic, microscopic and nanoscopic levels of organization. (Online version in colour.)
Mentions: Scanning ion conductance microscopy (SICM) invented by Hansma et al. [13] lately has been developed to image and analyse surface topography of live cells in our group [14–16]. SICM is a non-optical method that uses a nanopipette as a scanning probe to image cell surface structures with nanometre resolution [16]. SICM and a battery of associated innovative methods are unique among current imaging techniques, not only in spatial resolution, but also in the rich combination of imaging modalities with other functional and dynamic methods [10,17,18]. Recently, we have developed a hopping probe ion conductance microscopy (HPICM) [15], using a concept of ‘hopping’ from one imaging point to another, first implemented in SICM as the pulse mode back-step SICM mode [19]. Unlike previous attempts based on this concept, the HPICM managed to obtain nanoscale resolution in highly convoluted live cell samples without compromising the scan speed [15], and has already led to more elaborate techniques for single particle tracking [20] and functional imaging of receptor distribution [18]. The aim of this review is to describe the SICM technique alone or in combination with other optical and electrical methods to perform highly resolved dynamic and integrative analysis of cardiac structure, physiology and mechanisms of cardiovascular disease at the subcellular, cellular and tissue levels (figure 1).Figure 1.

Bottom Line: At the cellular level, heart failure leads to a pronounced loss of T-tubules in cardiac myocytes accompanied by a reduction in Z-groove ratio.The SICM pipette can be used for patch-clamp recordings of membrane potential and single channel currents.In conclusion, SICM provides a highly informative multimodal imaging platform for functional analysis of the mechanisms of cardiovascular diseases, which should facilitate identification of novel therapeutic strategies.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Science, National Heart and Lung Institute, Imperial College London, , Dovehouse Street, London SW36LY, UK.

ABSTRACT
Cardiovascular diseases are complex pathologies that include alterations of various cell functions at the levels of intact tissue, single cells and subcellular signalling compartments. Conventional techniques to study these processes are extremely divergent and rely on a combination of individual methods, which usually provide spatially and temporally limited information on single parameters of interest. This review describes scanning ion conductance microscopy (SICM) as a novel versatile technique capable of simultaneously reporting various structural and functional parameters at nanometre resolution in living cardiovascular cells at the level of the whole tissue, single cells and at the subcellular level, to investigate the mechanisms of cardiovascular disease. SICM is a multimodal imaging technology that allows concurrent and dynamic analysis of membrane morphology and various functional parameters (cell volume, membrane potentials, cellular contraction, single ion-channel currents and some parameters of intracellular signalling) in intact living cardiovascular cells and tissues with nanometre resolution at different levels of organization (tissue, cellular and subcellular levels). Using this technique, we showed that at the tissue level, cell orientation in the inner and outer aortic arch distinguishes atheroprone and atheroprotected regions. At the cellular level, heart failure leads to a pronounced loss of T-tubules in cardiac myocytes accompanied by a reduction in Z-groove ratio. We also demonstrated the capability of SICM to measure the entire cell volume as an index of cellular hypertrophy. This method can be further combined with fluorescence to simultaneously measure cardiomyocyte contraction and intracellular calcium transients or to map subcellular localization of membrane receptors coupled to cyclic adenosine monophosphate production. The SICM pipette can be used for patch-clamp recordings of membrane potential and single channel currents. In conclusion, SICM provides a highly informative multimodal imaging platform for functional analysis of the mechanisms of cardiovascular diseases, which should facilitate identification of novel therapeutic strategies.

Show MeSH
Related in: MedlinePlus