Separating cells:
Saved in:
| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
Oxford
BIOS Scientific Publ.
2001
New York Springer |
| Edition: | 1. publ. |
| Series: | The basics
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Includes bibliographical references and index |
| Physical Description: | VII, 168 S. graph. Darst. |
| ISBN: | 0387916121 1859961487 |
Staff View
MARC
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Record in the Search Index
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| adam_text | Contents
Abbreviations viii
Preface ix
Glossary xi
Chapter I Preparation of cell suspensions I
1. Diversity of cells 1
2. Why separate living cells? 2
3. Preparation of suspensions of single cells 2
4. Viability measurements 4
5. Separation of viable and nonviable cells 5
6. Characterization of cells 6
Protocol 1.1 Isolation of cells from bronchial alveolar lavage 14
Protocol 1.2 Isolation of cells from peritoneal lavage 15
Protocol 1.3 Isolation of cells from soft tissue by mechanical
disruption 16
Protocol 1.4 Preparation of vascular smooth muscle cells by
mechanical disruption and outgrowth in culture 17
Protocol 1.5 Preparation of endothelial cells from omental fat by
mechanical disruption and enzymatic digestion 18
Protocol 1.6 Isolation of neurons from rat ganglia by mechanical
disruption and enzymatic digestion 20
Protocol 1.7 Disaggregation of embryonic and normal and
malignant tissues 23
Protocol 1.8 Separation of renal proximal tubule cells by tissue
swelling and enzymatic digestion 25
Protocol 1.9 Preparation of rat hepatocytes by continuous
perfusion with enzyme solution 27
Protocol 1.10 Release of cultured cells from substratum by
trypsinization 29
Protocoll.il Isolation of yeast protoplasts (spheroplasts) 30
Protocol 1.12 Isolation of plant protoplasts 32
Protocol 1.13 Cell viability determined by trypan blue exclusion 34
Protocol 1.14 Cell viability determined by fluorescence staining
methods 36
Protocol 1.15 Cell viability determined by protein synthesis 37
Protocol 1.16 Cell viability determined by uptake of radiolabeled
thymidine 39
Protocol 1.17 Cell preparation by cytocentrifuge for immunostaining 41
Protocol 1.18 Characterization of cells by indirect
immunofluorescence staining 42
Protocol 1.19 Indirect (DAB) immunoperoxidase staining 44
Protocol 1.20 Metabolic characterization of endothelial cells 46
Chapter 2 Fractionation of cells by
sedimentation methods 49
1. Introduction 49
2. Separation media 50
3. Iso osmotic gradients 51
4. Choice of separation method 59
5. Separation of cells on the basis of size 59
6. Separation of cells on the basis of their density 60
7. Concluding remarks 68
Protocol 2.1 Isolation of platelets from human peripheral blood 70
Protocol 2.2 The isolation of mononuclear cells from diluted blood 71
Protocol 2.3 The isolation of mononuclear cells from whole blood 72
Protocol 2.4 The fractionation of human mononuclear cells 73
Protocol 2.5 The isolation of polymorphonuclear cells from blood 74
Protocol 2.6 Isolation of erythrocytes 75
Protocol 2.7 The isolation of viable spermatozoa from bovine
semen using Nycodenz gradients 76
Protocol 2.8 The isolation of viable spermatozoa from bovine
semen using OptiPrep gradients 77
Protocol 2.9 Purification and enrichment of viable cells prior to
further fractionation 78
Protocol 2.10 Isopycnic separation of rat liver cells using continuous
Percoll gradients 79
Protocol 2.11 Isolation of hepatocytes by differential pelleting 80
Protocol 2.12 Fractionation of sinusoidal cells 81
Protocol 2.13 The isolation and purification of plant protoplasts
using OptiPrep gradients 83
Protocol 2.14 Isolation of immunologically distinct cell
subpopulations using density perturbation methods 85
Chapter 3 Centrifugal elutriation 87
1. Introduction 87
2. Principles 88
3. Equipment 89
4. Conditions affecting separation 91
5. Applications of centrifugal elutriation 92
6. Advantages and disadvantages 98
Protocol 3.1 Standard procedure for cell separation by centrifugal
elutriation 101
Protocol 3.2 Separation of growth synchronous cells 103
Protocol 3.3 Purification of rat hepatocyte couplets 105
Chapter 4 Free flow electrophoresis 107
1. Introduction 107
2. Main types of FFE equipment 108
3. Buffers 109
4. Preparation of cell samples for FFE 111
5. FFE of cell samples 112
6. Factors affecting FFE 114
7. Applications of FFE 116
Protocol 4.1 Purification and preparation of lymphocytes for FFE 119
Protocol 4.2 Purification and preparation of platelets for FFE 121
Protocol 4.3 Purification and preparation of neutrophils for FFE 122
Protocol 4.4 Purification and preparation of kidney cells for FFE 123
Protocol 4.5 Purification and preparation of ascitic human cells
for FFE 124
Protocol 4.6 Re electrophoresis of FFE separated cells 125
Chapter 5 Affinity separation 127
1. Introduction 127
2. Separation by nonmagnetic techniques 127
3. Separation by magnetic techniques 129
Protocol 5.1 Isolation of monocytes by adherence to solid surfaces 138
Protocol 5.2 Isolation of HLA DR positive lymphocytes using
antibody antigen binding 139
Protocol 5.3 Direct positive selection of CD4* T cells from
whole blood 141
Protocol 5.4 Preparation of antibody coated cells for use in
indirect positive selection 143
Chapter 6 Flow cytometry 145
1. Introduction 145
2. The flow cytometer instrumentation 146
3. Types of flow sorting 150
4. Factors affecting cell sorting 151
5. Separation purity, yield and sterility 152
6. Applications 153
Protocol 6.1 Checklist for initial steps in setting up a flow sorter 156
Protocol 6.2 Guide to checking droplet delay time 157
Protocol 6.3 Checklist for measuring the yield of sorted cells 158
Protocol 6.4 Precautions taken to maintain sterility 159
List of manufacturers and suppliers 161
Index 165
|
| any_adam_object | 1 |
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| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.6078 |
| dewey-search | 571.6078 |
| dewey-sort | 3571.6078 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| edition | 1. publ. |
| format | Book |
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| id | DE-604.BV014759280 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T19:05:51Z |
| institution | BVB |
| isbn | 0387916121 1859961487 |
| language | English |
| lccn | 2002510819 |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009994303 |
| oclc_num | 45349873 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | VII, 168 S. graph. Darst. |
| publishDate | 2001 |
| publishDateSearch | 2001 |
| publishDateSort | 2001 |
| publisher | BIOS Scientific Publ. Springer |
| record_format | marc |
| series2 | The basics |
| spelling | Patel, D. Verfasser aut Separating cells Dipak Patel 1. publ. Oxford BIOS Scientific Publ. 2001 New York Springer VII, 168 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier The basics Includes bibliographical references and index Cell Separation methods Laboratory Manuals Cell separation Laboratory manuals Isolierung Chemie (DE-588)4122223-4 gnd rswk-swf Labortechnik (DE-588)4123602-6 gnd rswk-swf Zelle (DE-588)4067537-3 gnd rswk-swf Zelle (DE-588)4067537-3 s Isolierung Chemie (DE-588)4122223-4 s Labortechnik (DE-588)4123602-6 s DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009994303&sequence=000006&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Patel, D. Separating cells Cell Separation methods Laboratory Manuals Cell separation Laboratory manuals Isolierung Chemie (DE-588)4122223-4 gnd Labortechnik (DE-588)4123602-6 gnd Zelle (DE-588)4067537-3 gnd |
| subject_GND | (DE-588)4122223-4 (DE-588)4123602-6 (DE-588)4067537-3 |
| title | Separating cells |
| title_auth | Separating cells |
| title_exact_search | Separating cells |
| title_full | Separating cells Dipak Patel |
| title_fullStr | Separating cells Dipak Patel |
| title_full_unstemmed | Separating cells Dipak Patel |
| title_short | Separating cells |
| title_sort | separating cells |
| topic | Cell Separation methods Laboratory Manuals Cell separation Laboratory manuals Isolierung Chemie (DE-588)4122223-4 gnd Labortechnik (DE-588)4123602-6 gnd Zelle (DE-588)4067537-3 gnd |
| topic_facet | Cell Separation methods Laboratory Manuals Cell separation Laboratory manuals Isolierung Chemie Labortechnik Zelle |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009994303&sequence=000006&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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