Newly Developed Chemical Probes and Nano-Devices for Cellular Analysis

Abstract

Homeostasis in an organism is maintained by activities within and between cells. Therefore, one is required to analyze both the cellular activities within cells, such as metabolisms or signal transduction, and cellular interactions, such as ligand-receptor recognition. We have developed a cellular analyzing system including a "real-time cellular imaging system" and a "comprehensive analysis system for cellular responses" as shown in Fig. 1. The system has been established using newly developed chemical probes and nano-devices. As chemical probes, we have developed 1) molecular imaging probes, and 2) mass probes.As nano-devices, we built 1) nano electrochemical and optical probes, 2) a sputtered nano-film electrode, 3) nano-dot SPR, 4) a nano-dot target plate for MS, and 5) microfluidic devices. We will review each of these probes and devices, and finally introduce a "real-time cellular imaging system" with an application model using PC12 cells and a "comprehensive analysis system for cellular responses", which measured IL-1β (interleukin-1β) as an example. Chemical Probes 2·1 Molecular imaging probesFluorescent imaging based on fluorescent probes offers a highly sensitive, high-speed spatial and temporal analysis of cells, and is widely used in the fields of biology and physiology. A cellular analyzing system including a "real-time cellular imaging system" and a "comprehensive analyzing system for cellular responses" was developed. A "real-time cellular imaging system" is a system used to measure real-time imaging of multiple phenomena of a single cell with high special and temporal resolutions for the purpose to understand the pathology and physiology in a single cell and realize to single cell level diagnosis. A "real-time cellular imaging system" includes multi-probe imaging with AFM (atomic force microscopy), optical and SECM (scanning electrochemical microscopy) modes, which provides us with topological information and biochemical reactions at the local area of the interior and exterior of a cell. Scanning electrochemical/optical microscopy was applied to image PC12 cells. On the other hand, cells respond to their specific substances via their ligands. Therefore, the comprehensive analysis of protein-protein interaction is the important issue to determine the functions of cells. For this purpose, a "comprehensive analysis system for cellular responses" was developed. This system is based on SPR (surface plasmon resonance) and MS (mass spectrometry) using a nano-fabricated substrate. The interaction between IL-1β and anti-IL-1β antibodies was detected.

Current Application of Micro/Nano-Interfaces to Stimulate and Analyze Cellular Responses

“…6 Compared to other analytical methods such as fluorescence microscopy, confocal microscopy, and atomic force microscopy (AFM), SECM is more suitable for providing cellular biochemical information including cell responses to external stimuli, molecular transfer across cell membranes, or cell respiration along with cell morphology as reviewed by several articles reflecting this great amount of attention. 7,33,66,70,78 However, this review study deals exclusively with the recent academic achievement in the field of electrochemical characterization of single cells using SECM.…”

Nanobiodevice-based Single Biomolecule Analysis, Single-Cell Analysis, and in vivo Imaging for Cancer Diagnosis, Cancer Theranostics, and iPS Cell-based Regenerative Medicine

“…Since nanobiodevices have tremendous advantages, [1][2][3][4][5][6][7] they are applicable to single-cell analysis, [8][9][10] single biomolecular analysis 11,12 and the fast analysis of biomolecules developed by an appropriate small space with a short diffusion distance for cells and biomolecules, which have extremely small diffusion constants. 13 Nanomaterials, such as quantum dots, which have quantum confined effects, are essential to develop highly bright and long-life fluorescence materials, which will be indispensable for molecular imaging 14,15 and in vivo imaging.…”

“…[16][17][18] The research efforts in my laboratory have been focused on the development of novel nanobiodevices for biomedical applications. [1][2][3][4][5][6][7] In this review article, we mainly overview our research activities, including nanobiodevices for single-cell analysis, [20][21][22][23][24][25] single biomolecule analysis, 19,[26][27][28][29]41,46,50,[54][55][56][57][58] biomarker detections, 23,[30][31][32][33]39,[47][48][49] cancer theranostics, and iPS cell in vivo imaging. [20][21][22][23][24][25][34][35][36][37][38]…”

Ionic Probe Attachment Ionization Mass Spectrometry

“…5 erefore, an e ective adjustable-charge-state ionization technique 6 is required for these molecules. We have recently developed a new method called ionic probe attachment ionization .…”