Practical Cell Analysis

As analytical chemistry and biology move closer together, biologists are performing increasingly sophisticated analytical techniques on cells. Chemists are also turning to cells as a relevant and important sample to study newly developed methods. Practical Cell Analysis provides techniques, hints, and time-saving tips explaining what may be “common knowledge” to one field but are often hidden or unknown to another. Within this practical guide: The procedures and protocols for cell separation, handling cells on a microscope and for using cells in microfluidic devices are presented. Elements of cell culture are taken and combined with the practical advice necessary to maintain a cell lab and to handle cells properly during an analysis The main chapters deal with the fundamentals and applied aspects of each technique, with one complete chapter focusing on statistical considerations of analyzing cells Many diagram-based protocols for some of the more common cell processes are included Chapter summaries and extensive tables are included so that key information can be looked up easily in the lab setting Much like a good manual or cookbook this book is a useful, practical guide and a handy reference for all students, researchers and practitioners involved in cellular analysis.

The selective combination of physical, biochemical, and immunological prin ciples, along with new knowledge concerning the biology of cells and advance ments in engineering and computer sciences, has made possible the emergence of highly sophisticated and powerful methods for the analysis of cells and their constituents. This series on Cell Analysis is, therefore, aiming at providing the theoretical and practical background on how these methods work and what kind of information can be obtained. Cell Analysis will cover techniques on cell separation, cell identification and classification, characterization of orga nized cellular components, functional properties of cells, and cell interactions. Applications in cell biology, immunology, genetics, toxicology, specific diseases, diagnostics and therapeutics, and other areas will be covered whenever relevant results exist. Nicholas Catsimpoolas Boston, Massachusetts vii Contents Chapter I Quantification of Red Blood Cell Morphology James W. Bacus I. History .. II. Details of Red Cell Measurements. 3 III. Cell Sample Population Distributions. 11 IV. Discussion and Summary. 25 References. 30 Chapter 2 Laser Microirradiation and Computer Video Optical Microscopy in Cell Analysis Michael W. Berns and Robert J. Walter I. Introduction 33 II. Laser Microbeams 34 III. Computer-Enhanced Video Microscopy for Laser Microsurgery.

This volume explores the latest advancements and techniques used to study cell analysis, their capabilities, and the type of results that can be obtained. The chapters in this book cover topics such as FACS; fluorescence microscopy; organic spectroscopy such as MALDI; inorganic spectroscopy such as ICP-MS; and sequencing. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Single Cell Analysis: Methods and Protocols is a valuable tool for any researcher interested in learning more about this important and developing field.

This volume highlights the most interesting biomedical and clinical applications of high-dimensional flow and mass cytometry. It reviews current practical approaches used to perform high-dimensional experiments and addresses key bioinformatic techniques for the analysis of data sets involving dozens of parameters in millions of single cells. Topics include single cell cancer biology; studies of the human immunome; exploration of immunological cell types such as CD8+ T cells; decipherment of signaling processes of cancer; mass-tag cellular barcoding; analysis of protein interactions by proximity ligation assays; Cytobank, a platform for the analysis of cytometry data; computational analysis of high-dimensional flow cytometric data; computational deconvolution approaches for the description of intracellular signaling dynamics and hyperspectral cytometry. All 10 chapters of this book have been written by respected experts in their fields. It is an invaluable reference book for both basic and clinical researchers.

This detailed volume serves as a collection of methods for single-cell protein analysis, created by combining different protocols, taking advantage of new emerging technologies, and improving upon conventional methods to guide researchers aiming to perform protein analysis in single cells. Ranging from simple to complex, conventional to the most current technologies, these chapters offer readers the ability to choose the best suited methodologies for them, based on the sample type and the available technologies or equipment. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Single-Cell Protein Analysis: Methods and Protocols aims to help researchers utilizing single-cell protein analysis in their studies as well to inspire the development of the next generation of improved protein analysis methods in single cells.

The analysis of blood, bone marrow and tissue fluid specimens requires a multi-faceted approach with the integration of scientific data from a number of disciplines. No single discipline can operate in isolation or errors will occur. Flow cytometry is in a privileged position in that it can provide rapid analysis of specimens and it is often the first definitive investigation to produce results and help formulate a working diagnosis. This companion text to Practical Flow Cytometry in Haematology Diagnosis contains 100 worked examples drawn from real clinical cases presenting to the authors’ institution. Cases are illustrated with peripheral blood and bone marrow cytology, tissue pathology and cytogenetic and molecular data, which are integrated to generate, where appropriate, a diagnosis based on the WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. The spectrum of clinical cases includes adult and paediatric patients, and both neoplastic and reactive disorders. The cases appear in no particular order to challenge the reader to make their own diagnosis. The reader will review May−Grünwald−Giemsa (MGG)-stained films of peripheral blood and bone marrow aspirates presented alongside flow cytometric data and haematoxylin and eosin (H&E)-stained bone marrow and other tissue biopsy sections. Immunohistochemistry is used to further clarify the tissue lineage and cell differentiation. Cytogenetic studies using metaphase preparations are used to identify translocations and chromosome gains and losses whilst interphase fluorescence in situ hybridisation (FISH) studies and polymerase chain reaction (PCR) are used to identify gene fusions, gene rearrangements and deletions. Each case concludes with a discussion of the features that are important to making a diagnosis. The cases are also listed according to disease classification in the appendix so that the text can also be used as a reference. Practical Flow Cytometry in Haematology: 100 Worked Examples: Provides a practical, example-based resource for flow cytometry Demonstrates how flow cytometry results should be interpreted and applied to optimize patient care Includes both malignant and benign conditions Can be used in conjunction with Practical Flow Cytometry in Haematology Diagnosis, by the same author team (ISBN 9780470671207) Practical Flow Cytometry in Haematology: 100 Worked Examples is ideal for practicing haematologists and histopathologists with an interest in haematopathology, but particularly directed at trainee haematologists and scientists preparing for FRCPath and related examinations.

This volume highlights the most interesting biomedical and clinical applications of high-dimensional flow and mass cytometry. It reviews current practical approaches used to perform high-dimensional experiments and addresses key bioinformatic techniques for the analysis of data sets involving dozens of parameters in millions of single cells. Topics include single cell cancer biology; studies of the human immunome; exploration of immunological cell types such as CD8+ T cells; decipherment of signaling processes of cancer; mass-tag cellular barcoding; analysis of protein interactions by proximity ligation assays; Cytobank, a platform for the analysis of cytometry data; computational analysis of high-dimensional flow cytometric data; computational deconvolution approaches for the description of intracellular signaling dynamics and hyperspectral cytometry. All 10 chapters of this book have been written by respected experts in their fields. It is an invaluable reference book for both basic and clinical researchers.