Tying together technological and conceptual advances of the past decade, this seminal book introduces the novel approach of chemical cytometry -- the biochemical analysis of cell constituents at the single cell level. As such, it combines recently developed methods for the handling of single cells, the separation of their components, and the detection and analysis of biomolecular species with application examples from cell biology, neurobiology and molecular medicine. The editor has succeeded in acquiring contributions from the recognized leaders in the field, resulting in a complete and authoritative overview for both developers as well as users of these new methods.
Chang Lu is an associate professor of chemical engineering at Virginia Tech, Blacksburg (USA). From 2004 to 2009 he was an assistant and associate professor of biological engineering at Purdue University at West Lafayette. He obtained his academic degrees from Peking University and from the University of Illinois at Urbana-Champaign, and received post-doctoral training at Cornell University. His research has been focused on Microfluidic devices for single cell analysis and tissue engineering. Professor Lu has received a number of awards, including the NSF Career Award and the Wallace H. Coulter foundation Early Career Award in Biomedical Engineering.
Preface ORIGIN, CURRENT STATUS, AND FUTURE PERSPECTIVES OF CHEMICAL CYTOMETRY The Cell and Cytometry Flow and Image Cytometry Chemical Cytometry Chemical Cytometry of DNA and mRNA Metabolic Cytometry Future Perspectives on Instrumentation for Chemical Cytometry METABOLIC CYTOMETRY - THE STUDY OF GLYCOSPHINGOLIPID METABOLISM IN SINGLE PRIMARY CELLS OF THE DORSAL ROOT GANGLIA Introduction Material and Methods Results and Discussion Conclusions CELL SIGNALING STUDIES AT THE SINGLE-CELL LEVEL Introduction Analytes Examined and Reporters Used Cell Preparation and Reporter Loading Cell Lysis and Sampling Techniques Electrophoresis Separation Conditions Detection Automation and Throughput ULTRASENSITIVE DETECTION OF LOW-COPY-NUMBER MOLECULES FROM SINGLE CELLS Introduction Microchip Designs for Single-Cell Analysis and/or Cell Manipulation Ultrasensitive Detection Methods for Single-Cell Analysis Single-Cell Analysis with Single-Molecule Sensitivity on Integrated Microfluidic Chip Conclusions CAPILLARY ELECTROPHORESIS OF NUCLEIC ACIDS AT THE SINGLE-CELL LEVEL Introduction On-Line Cell Analysis Direct Gene and Gene Expression Analysis Without Amplification Potential Alternative Techniques for Single-Cell Gene and Gene Expression Analysis Conclusions MICROFLUIDIC TECHNOLOGY FOR SINGLE-CELL ANALYSIS Introduction Biological Significance of Single-Cell Analysis Microfluidic Devices in Our Laboratories Materials, Methods, and Protocols Conclusions ON-CHIP ELECTROPORATION AND ELECTROFUSION FOR SINGLE-CELL ENGINEERING Introduction Single-Cell Electroporation in Microfluidic Devices Single-Cell Electrofusion in Microfluidic Devices Conclusions ELECTROPORATIVE FLOW CYTOMETRY FOR SINGLE-CELL ANALYSIS Introduction Flow-Through Electroporation under Constant Voltage Electroporative Flow Cytometry for Detecting Protein Translocation Electroporative Flow Cytometry for Measuring Single-Cell Biomechanics Electroporative Flow Cytometry for Selectively Releasing and Analyzing Specific Intracellular Molecules Conclusion ULTRASENSITIVE ANALYSIS OF INDIVIDUAL CELLS VIA DROPLET MICROFLUIDICS Introduction Droplet Properties Droplet Generation Cell Encapsulation Droplet Manipulation Droplet Concentration Control Temperature Control of Droplets Detection in Droplets Conclusions PROBING EXOCYTOSIS AT SINGLE CELLS USING ELECTROCHEMISTRY Introduction Measurement Requirements Electrode Fabrication Measurements at Single Cells Fusion Pore Dynamics Conclusions ELECTROCHEMICAL DETERMINATION OF ENZYME ACTIVITY IN SINGLE CELLS Introduction Electrochemical Detection Coupled with Capillary Electrophoresis Voltammetry Scanning Electrochemical Microscopy (SECM) High-Throughput ECD Perspective SINGLE-CELL MASS SPECTROMETRY Introduction Mass Spectrometry Overall Outlook for Single-Cell MS OPTICAL SENSING ARRAYS FOR SINGLE-CELL ANALYSIS Introduction to Fiber-Optic Single-Cell Arrays Advantages of Fiber-Optic Single-Cell Arrays Fiber-Optic Arrays Singe-Cell Arrays for Bacteria Single-Cell Arrays for Yeast Single-Cell Arrays for Mammalian Cells Image and Data Analysis for Single-Cell Arrays Summary