Methods of Cell Separation: (Biological Separations)

Presently, the need for methods involving separation, identification, and characterization of different kinds of cells is amply realized among immu- nologists, hematologists, cell biologists, clinical pathologists, and cancer researchers. Unless cells exhibiting different functions and stages of differ- entiation are separated from one another, it will be exceedingly difficult to study some of the molecular mechanisms involved in cell recognition, spe- cialization, interactions, cytotoxicity, and transformation. Clinical diagno- sis of diseased states and use of isolated cells for therapeutic (e. g. , immu- notherapy) or survival (e. g, transfusion) purposes are some of the pressing areas where immediate practical benefits can be obtained by applying cell separation techniques. However, the development of such useful methods is still in its infancy. A number of good techniques exist based either on the physical or biological properties of the cells, and these have produced some valuable results. Still others are to be discovered. Therefore, the purpose of this open-ended treatise is to acquaint the reader with some of the basic principles, instrumentation, and procedures presently in practice at various laboratories around the world and to present some typical applications of each technique to particular biological problems.

Table of Contents:
1 Computer Analysis of Lymphocyte Images.- I. Introduction.- A. Computable Image Information.- B. Chromatin Distribution Patterns.- II. Selection of Equipment.- A. The Microscope Photometer.- B. Interface.- C. Computer.- D. Photomicrography and Video-Recording.- III. Computer Programs.- A. Design Considerations.- B. Data Staging and File Handling.- IV. Analytical Tasks.- A. Cell Image Features.- B. Statistical Evaluation of the Data.- C. Feature Evaluation and Selection.- D. Supervised Learning Algorithms.- E. Unsupervised Learning Algorithms.- V. Application of Image Analysis to Biological Problems.- A. X-Ray Radiation: Study One.- B. X-Ray Radiation: Study Two.- C. X-Ray Radiation: Study Three.- D. Virus-Altered Cells: Study Four.- E. Detection of T and B Cells: Study Five.- References.- 2 Principles of Continuous Flow Cell Separation in a Circumferential Flow Disposable Channel.- I. Introduction.- II. Design Concepts.- III. Channel Description.- IV. Channel Operation.- A. Results.- V. Channel Performance.- A. Granulocyte Collection.- B. Mononuclear Cell Collection.- C. Plasma Exchange.- VI. Summary.- References.- 3 Magnetic Microspheres in Cell Separation.- I. Background.- A. Physics of Magnetophoresis.- B. Previous Examples.- II. Procedures.- A. Preliminary.- B. Preparation of Single-Domain Magnetic Affinity Beads.- III. Examples.- A. Toxic Lectins.- B. Antibody Ligands.- References.- 4 Isolation of Human Blood Phagocytes by Counterflow Centrifugation Elutriation.- I. Introduction.- II. Isolation of Granulocytes from Human Blood.- III. Isolation of Human Blood.- A. Isolation of Monocytes by Incremental Flow Elutriation.- IV. Appraisal of Counterflow Centrifugation for the Isolation of Leukocytes.- A. Who Needs 1 x 108 Granulocytes?.- B. Scaling the Numbers to 1 x 1010 Granulocytes.- C. Some Technical Considerations.- References.- 5 Biological Methods for the Separation of Lymphoid Cells.- I. General Considerations.- II. Rosetting Methods.- A. Introduction.- B. Spontaneous Rosettes of Sheep Erythrocytes with Human T Lymphocytes.- C. Separation of Human Lymphocyte Subpopulations Bearing Fc Receptors.- D. Separation of Lymphocyte Subpopulations Bearing Complement Receptors.- E. Separation of Human B Lymphocyte Subpopulations by Rosetting Methods.- F. Separation of Rodent T and B Lymphocytes by Rosetting Methods.- G. Other Spontaneous Erythrocyte Rosettes.- III. Affinity Chromatography of Cells and Cellular Immunoabsorbent Methods.- A. Introduction.- B. Separation of Immunoglobulin-Positive and Immunoglobulin-Negative Lymphocytes by Affinity Chromatography Methods.- C. Separation of Fc-Receptor-Bearing Lymphocytes by Affinity Methods.- D. Separation of Specific Antigen-Binding Lymphocytes by Affinity Methods.- E. Separation of Cytotoxic T Lymphocytes by Cellular Immunoabsorbent Methods.- IV. Elimination of Lymphocyte Subpopulations by Antisera Plus Complement Treatment.- V. Elimination of Specific Lymphocyte Subpopulations by Selective Incorporation of High-Specific-Activity Radioisotopes ("Suicide" Methods).- VI. Separation of Lymphocyte Subpopulations by the Use of Lectins.- VII. Conclusions.- References.