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Abstract

Chapter I. Force Spectroscopy for Biomolecular Analysis Using Atomic Force Microscopy Investigation of molecular interaction force, especially at single molecule level is very important to understand biomolecular phenomena and mechanisms. Atomic force microscopy (AFM) is a versatile tool that can image the topography of surfaces and quantify the physical properties of molecular interactions. In this chapter, surface modification methods for single molecule force spectroscopy are described. Utilizing specific surface modification methods, AFM can detect and analyze various biomolecules in depth. AFM can provide image of samples at the nanometric resolution as well as reveal location and distribution of a specific molecule on surface. Chapter II. Quantification of a Neurological Protein in a Single Cell Without Signal Amplification Proteins are key biomolecules that not only play various roles in a living body, but are also frequently used as biomarkers. If these proteins can be quantified, in particular, those in a single cell can be addressed, understanding role of the proteins will be deepened, and diagnosing diseases and abnormality will be further upgraded. In this study, we quantified a neurological protein in a single cell using atomic force microscopy (AFM). After capturing specifically disrupted in schizophrenia 1 (DISC1) in a single cell onto a microspot immobilizing the corresponding antibody on the surface, force-mapping with AFM was followed to visualize individual DISC1. Although a large variation of the number of DISC1 in a cell was observed, the average number is 4.38 × 10^3, and the number agrees with the ensemble-averaged value. The current AFM approach for the quantitative analysis of proteins in a single cell should be useful to study molecular behavior of proteins in depth and to follow physiological change of individual cells in response to external stimuli.