Bioimaging

 
 Bioimaging
 Background
Molecular recognition between receptors and ligands plays a crucial role in life because the cellular behaviors, the immune systems and many types of diseases depend on this process. New micromanipulation techniques combining highly sensitive force measurement and accurate positioning with nanometer precision have offered researchers the opportunities to directly manipulate individual biological macromolecules such as nucleic acids and proteins, and to measure both intermolecular and intramolecular forces with high accuracy. The atomic force microscopy (AFM) is a well-established technique capable of quantification of biomolecular interactions at the single-molecule level with piconewton (pN) sensitivity. We are developing a novel method to detect mRNA distribution in cells or tissues using an AFM modified with a single-molecule probe technology. Until now, no simple method has been available for analyzing mRNA levels in cells or tissues without complex processes including in situ hybridization and immunological methods. Our strategy enables one to examine mRNA distribution in individual cells or small tissues with single-molecule sensitivity and high spatial resolution. To use AFM for the study of molecular recognition, the immobilization scheme for attaching biomolecules onto an AFM tip is critical for the success of an experiment. We modify our AFM tip using dendrons, cone-shaped synthetic molecules, which is expected to provide a simple, quick and reliable method for determining gene expression levels within a single cell.
 Contents
  -Development of molecular imaging technique for the analysis of mRNA distributions with single-molecule sensitivity and
high spatial resolution
-Development of novel imaging method using multimodal probes to assay the topology of protein molecules
-Improvement of the existing SECM-AFM (Electrochemical Microscopy – Atomic Force Microscopy) nanoprobe and
its biological application
-Development of new single cell RNA counting method using microfluidic devices
-Investigation of RNA-based biological networks and the functional role of RNAs in aging
Molecular imaging technique for the analysis of mRNA distributions with single-molecule sensitivity and high spatial resolution in animal cell. a, Expression of the Pax6 mRNA in a coronal section of a mouse embryonic brain. B, A higher resolution map of distribution of Pax6 mRNA (Nucleic Acids Res 37(2), e10, 2009).
Molecular imaging strategy using a dendron-modified AFM tip for the analysis of mRNA distributions with single-molecule sensitivity in plant cell.
 Member for these studies
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