Chemical Cytometry Ultrasensitive Analysis Of Single Cells

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, West Lafayette (USA). 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.
List of Contributors.
1 Origin, Current Status, and Future Perspectives of Chemical Cytometry (Norman J. Dovichi).
1.1 The Cell and Cytometry.
1.2 Flow and Image Cytometry.
1.3 Chemical Cytometry.
1.4 Chemical Cytometry of DNA and mRNA.
1.5 Metabolic Cytometry.
1.6 Future Perspectives on Instrumentation for Chemical Cytometry.
Acknowledgments.
References.
2 Metabolic Cytometry – The Study of Glycosphingolipid Metabolism in Single Primary Cells of the Dorsal Root Ganglia (Colin D. Whitmore, Jillian Prendergast, David Essaka, Ole Hindgaul, Monica P. Palcic, Ronald L. Schnaar, and Norman J. Dovichi).
2.1 Introduction.
2.2 Material and Methods.
2.3 Results and Discussion.
2.4 Conclusions.
Acknowledgments.
References.
3 Cell Signaling Studied at the Single-cell Level (Angela Proctor, Shan Yang, Sumith Kottegoda, Michael Brenner, Ryan M. Phillips, Christopher E. Sims, and Nancy L. Allbritton).
3.1 Introduction.
3.2 Analytes Examined and Reporters Used.
3.3 Cell Preparation and Reporter Loading.
3.4 Cell Lysis and Sampling Techniques.
3.5 Electrophoresis Separation Conditions.
3.6 Detection.
3.7 Automation and Throughput.
References.
4 Ultrasensitive Detection of Low-copy-number Molecules from Single Cells (Kangning Ren and Hongkai Wu).
4.1 Introduction.
4.2 Microchip Designs for Single-cell Analysis and/or Cell Manipulation.
4.3 Ultrasensitive Detection Methods for Single-cell Analysis.
4.4 Single-cell Analysis with Single-molecule Sensitivity on Integrated Microfluidic Chip.
4.5 Conclusions.
References.
5 Capillary Electrophoresis of Nucleic Acids at the Single-cell Level (Ni Li and Wenwan Zhong).
5.1 Introduction.
5.2 On-line Cell Analysis.
5.3 Direct Gene and Gene Expression Analysis Without Amplification.
5.4 Potential Alternative Techniques for Single-cell Gene and Gene Expression Analysis.
5.5 Conclusions.
References.
6 Microfluidic Technology for Single-cell Analysis (Yan Chen and Jiang F. Zhong).
6.1 Introduction.
6.2 Biological Significance of Single-cell Analysis.
6.3 Microfluidic Devices in Our Laboratories.
6.4 Materials, Methods, and Protocols.
6.5 Conclusions.
References.
7 On-chip Electroporation and Electrofusion for Single-cell Engineering (Ana Valero and Albert van den Berg).
7.1 Introduction.
7.2 Single-cell Electroporation in Microfluidic Devices.
7.3 Single-cell Electrofusion in Microfluidic Devices.
7.4 Conclusions.
References.
8 Electroporative Flow Cytometry for Single-cell Analysis (Chang Lu, Jun Wang, Ning Bao, and Hsiang-Yu Wang).
8.1 Introduction.
8.2 Flow-through Electroporation under Constant Voltage.
8.3 Electroporative Flow Cytometry for Detecting Protein Translocation.
8.4 Electroporative Flow Cytometry for Measuring Single-cell Biomechanics.
8.5 Electroporative Flow Cytometry for Selectively Releasing and Analyzing Specific Intracellular Molecules.
8.6 Conclusions.
Acknowledgments.
References.
9 Ultrasensitive Analysis of Individual Cells via Droplet Microfluidics (Robert M. Lorenz and Daniel T. Chiu).
9.1 Introduction.
9.2 Droplet Properties.
9.3 Droplet Generation.
9.4 Cell Encapsulation.
9.5 Droplet Manipulation.
9.6 Droplet Concentration Control.
9.7 Temperature Control of Droplets.
9.8 Detection in Droplets.
9.9 Conclusions.
References.
10 Probing Exocytosis at Single Cells Using Electrochemistry (Yan Dong, Michael L. Heien, Michael E. Kurczy, and Andrew G. Ewing).
10.1 Introduction.
10.2 Measurement Requirements.
10.3 Electrode Fabrication.
10.4 Measurements at Single Cells.
10.5 Fusion Pore Dynamics.
10.6 Conclusions.
Acknowledgments.
References.
11 Electrochemical Determination of Enzyme Activity in Single Cells (Wenrui Jin).
11.1 Introduction.
11.2 Electrochemical Detection Coupled with Capillary Electrophoresis.
11.3 Voltammetry.
11.4 Scanning Electrochemical Microscopy (SECM).
11.5 High-throughput ECD.
11.6 Perspective.
Acknowledgment.
References.
12 Single-cell Mass Spectrometry (Ann Knolhoff, Stanislaw Rubakhin, and Jonathan V. Sweedler).
12.1 Introduction.
12.2 Mass Spectrometry.
12.3 Overall Outlook for Single-cell MS.
Acknowledgments.
References.
13 Optical Sensing Arrays for Single-cell Analysis (Ragnhild D. Whitaker and David R. Walt).
13.1 Introduction to Fiber-optic Single-cell Arrays.
13.2 Advantages of Fiber-optic Single-cell Arrays.
13.3 Fiber-optic Arrays.
13.4 Single-cell Arrays for Bacteria.
13.5 Single-cell Arrays for Yeast.
13.6 Single-cell Arrays for Mammalian Cells.
13.7 Image and Data Analysis for Single-cell Arrays.
13.8 Summary.
References.
Index.