Protein localization by fluorescence microscopy: a practical approach
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Oxford [u.a.]
Oxford Univ. Press
2000
|
| Series: | The practical approach series
218 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XXXIV, 231 S. Ill., graph. Darst. |
| ISBN: | 0199637407 0199637415 |
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Record in the Search Index
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| adam_text | PROTEIN LOCALIZATION BY FLUORESCENCE MICROSCOPY A PRACTICAL APPROACH
EDITED BY VICTORIA J. ALLAN SCHOOL OF BIOLOGICAL SCIENCES, UNIVERSITY OF
MANCHESTER, 2.205 STOPFORD BUILDING, OXFORD ROAD, MANCHESTER ML 3 9PT
TECHNISCHE UNIVERSITAT DARMSTADT FACHBEREICH 10 * BIOLOGIE * BIBLIOTHEK
* SCHNITTSPAHNSTRAFLE 10 D-64287 DARMSTADT INV.-NR. OXPORD UNIVERSITY
PRESS CONTENTS LIST OF CONTRIBUTORS ABBREVIATIONS 1. BASIC
IMMUNOFLUORESCENCE VICTORIA I. ALLAN 1. INTRODUCTION IMPORTANT
CONSIDERATIONS 2. GROWING CELLS ON COVERSLIPS 3. FIXATION SOLVENT
FIXATION FIXATION BY CHEMICAL CROSS-LINKING ALDEHYDE CROSS-LINKING OTHER
CROSS-LINKERS 4. PERMEABILIZATION PRE-EXTRACTION BEFORE FIXATION 5.
LABELLING PROTOCOLS ANTIBODY QUALITY AND PREPARATION SINGLE VERSUS
MULTIPLE LABELLING MOUNTING COVERSLIPS FOR MICROSCOPY 6. CONTROLS 7.
DOUBLE LABELLING AS A MEANS FOR LOCALIZING YOUR PROTEIN WITHIN THE CELL
CYTOSKELETAL STRUCTURES MEMBRANOUS ORGANELLES ENDOPLASMIC RETICULUM
GOLGI APPARATUS, ENDOSOMES AND LYSOSOMES MITOCHONDRIA ACKNOWLEDGEMENTS
REFERENCES 1 1 2 4 7 8 8 11 12 14 15 16 17 18 20 21 21 21 21 23 23 25 25
2. IMMUNOFLUORESCENT LABELLING OF SECTIONS 27 ALISON J. NORTH AND J.
VICTOR SMALL 1. INTRODUCTION 27 CONTENTS 2. CHOICE OF SECTIONING
PROCEDURE 28 CONVENTIONAL (5-10 JIM) SECTIONS 28 SEMI-THIN (0.1-1 |IM)
SECTIONS 28 THE PROS AND CONS 28 METHODS OF TISSUE PREPARATION FOR
SEMI-THIN SECTIONING 29 -3. TISSUE FIXATION 32 PRINCIPLES OF FIXATION
AND THE IMMUNOCYTOCHEMICAL COMPROMISE 32 ^ PREPARATION OF BUFFERED
FIXATIVE SOLUTIONS 33 FIXATION AND PROCESSING FOR ALDEHYDE-SENSITIVE
ANTIGENS * 34 4. PREPARATION OF CONVENTIONAL (5-10 UM) SECTIONS 34 5.
PREPARATION OF SEMI-THIN (0.1-1 UM) SECTIONS 34 PROCESSING TISSUE 34
PROCESSING SINGLE CELLS OR CELLULAR AGGREGATES 35 SECTION THICKNESS 36
* HANDLING SEMI-THIN SECTIONS 37 SECTIONING BY THE TOKUYASU METHOD 38
CRYOPROTECTION 38 MOUNTING AND FREEZING 38 SECTIONING 38 SECTION
RETRIEVAL 39 SECTIONING OF PVA-EMBEDDED MATERIAL 40 EMBEDDING IN PVA ,
40 SECTIONING 40 6. IMMUNOFLUORESCENT LABELLING 42 LABELLING PROCEDURE
.. 42 PRIMARY AND SECONDARY ANTIBODIES 42 , OPTIMIZATION OF LABELLING
CONDITIONS 42 CONTROLS 43 STRATEGIES FOR MULTIPLE IMMUNOLABELLING 43 7.
AUTOFLUORESCENCE 45 8. ANTI-FADING MOUNTANTS 46 9. CONCLUDING REMARKS 46
REFERENCES 47 3. SIMULTANEOUS IN SITU DETECTION OF DNA AND PROTEINS 51
KLAUS ERSFELD AND ELISA M. STONE 1. INTRODUCTION 51 2. PREPARATION OF
LABELLED DNA PROBES 51 LABELLING OF DNA BY NICK TRANSLATION 52 XIV
CONTENTS LABELLING OF DNA BY PCR 54 3 -LABELLING OF OLIGONUCLEOTIDES
FOR FISH PROBES 55 3. PREPARATION OF CELLS 56 4. COMBINED
IMMUNOFLUORESCENCE AND IN SITU HYBRIDIZATION 58 5. APPLICATIONS OF
COMBINED IMMUNOFLUORESCENCE AND FISH IN YEAST 60 CO-LOCALIZING
CHROMOSOMAL PROTEINS AND CHROMOSOMAL DOMAINS 60 CO-LOCALIZING
CYTOSKELETAL PROTEINS AND CHROMOSOMAL DOMAINS 62 6. MICROSCOPY AND IMAGE
ANALYSIS 64 ACKNOWLEDGEMENTS R 65 REFERENCES 65 4. INSTRUMENTS FOR
FLUORESCENCE IMAGING 67 W. B. AMOS 1. INTRODUCTION 67 2. LIGHT BUDGET 70
3. COMPARING INSTRUMENTS 71 QUANTUM EFFICIENCY 71 NOISE 73 DYNAMIC
RANGE 73 PHOTON COUNTING 74 4. GETTING THE RIGHT MAGNIFICATION 75
MAGNIFICATION FOR CAMERAS 75 MAGNIFICATION IN LASER-SCANNING MICROSCOPES
76 5. LIGHT SOURCES AND FLUOROCHROMES 77 6. LIGHT DETECTORS 78
PHOTOMULTIPLIERS 78 PHOTODIODES 80 AVALANCHE PHOTODIODES 81 TELEVISION
CAMERAS , 81 VIDICON TUBE 82 SIT CAMERAS 82 CCD CAMERAS 82 VIDEO-RATE
CCDS AND READOUT NOISE 84 LONG INTEGRATION CCDS 85 ICCDS 85 CMOS CAMERAS
87 CAMERA-BASED SPECTRAL IMAGING 87 TESTING CAMERAS 88 7. SCANNING
OPTICAL MICROSCOPES 88 ADVANTAGES OF SCANNING PER SE 88 XV 1 CONTENTS
CONFOCAL MICROSCOPES 89 CONFOCAL MICROSCOPES WHERE A SINGLE SPOT OF
LIGHT IS SCANNED 89 ACOUSTO-OPTIC SCANNING 93 PARALLEL CONFOCAL
MICROSCOPES (MULTI-SPOT OR SLIT SCANNERS) 94 HOW TO TEST A CONFOCAL
MICROSCOPE 97 CHOOSING AN OBJECTIVE LENS 97 DOES A CONFOCAL MICROSCOPE
HAVE BETTER RESOLUTION THAN A CONVENTIONAL ONE? 98 HOW ARE THE LATERAL
AND AXIAL RESOLUTIONS CHANGED BY CONFOCAL OPERATION? - * 9 9 WHAT CAN
BE DONE ABOUT CHROMATIC ABERRATION IN CONFOCAL IMAGING? 101 MULTIPHOTON
IMAGING 102 HOW DO MULTIPHOTON OPTICS DIFFER FROM CONFOCAL? 102
APPARATUS FOR MULTIPHOTON IMAGING 104 PROS AND CONS OF MULTIPHOTON
VERSUS CONFOCAL IMAGING 104 8. HOW TO CHOOSE A FLUORESCENCE IMAGING
SYSTEM 105 ACKNOWLEDGEMENTS 106 REFERENCES 106 5. FLUORESCENCE
MICROSCOPY OF LIVING VERTEBRATE CELLS 109 RAINER PEPPERKOK AND DAVID
SHIMA 1. INTRODUCTION 109 .2. HARDWARE AND SOFTWARE REQUIREMENTS 110
MICROSCOPES AND ASSOCIATED HARDWARE 111 3D SECTIONING MICROSCOPE 111
CONFOCAL LASER-SCANNING MICROSCOPES 112 IMAGING DETECTORS FOR 3D
SECTIONING MICROSCOPY 112 CCD CAMERAS 113 INTENSIFIED CCD CAMERAS 115
COLOUR CAMERAS 115 TEMPERATURE AND ENVIRONMENTAL CONTROL 116 DATA
ANALYSIS, IMAGE PROCESSING AND DATA PRESENTATION 116 3. FLUORESCENT
LABELLING OF MOLECULES AND ORGANELLES IN LIVING CELLS 117 CELL-PERMEABLE
MARKERS 117 LABELLING BY MICRO-INJECTION OF FLUORESCENTLY LABELLED
PROTEINS OR ANTIBODIES 117 FLUORESCENT LABELLING OF PROTEINS 118
MICRO-INJECTION OF FLUORESCENTLY LABELLED MARKERS 121 4. UTILIZING GFP
AND ITS VARIANTS IN LIVE-CELL FLUORESCENCE MICROSCOPY 123 XVI CONTENTS
INTRODUCTION 123 PRACTICAL CONSIDERATIONS 123 CREATING GFP CHIMERAS 125
CONSIDERATIONS FOR STABLE AND TRANSIENT EXPRESSION OF GFP CHIMERAS IN
CELLS 126 , 5. EXAMPLES OF IMAGING FLUORESCENT MOLECULES IN LIVING CELLS
127 REAGENTS AND MICROSCOPY SET-UP 127 MITOSIS 128 R VISUALIZING
MEMBRANE TRANSPORT IN THE SECRETORY PATHWAY 129 REFERENCES 132 6.
VISUALIZING FLUORESCENCE IN DROSOPHILA * OPTIMAL DETECTION IN THICK
SPECIMENS 133 ILAN DAVIS T 1. INTRODUCTION 133 2. SPREAD OF LIGHT FROM A
POINT SOURCE 134 BASIC PROPERTIES OF OBJECTIVE LENSES 134 CLASSES OF
OBJECTIVE LENS 135 RESOLUTION AND BRIGHTNESS OF OBJECTIVE LENSES 135
OPTICAL ABERRATIONS AND THEIR CORRECTION 136 MEASURING THE SPREAD OF
LIGHT WITHIN A SPECIMEN 139 DETERMINING THE POINT-SPREAD FUNCTION OF A
PARTICULAR LENS AND MICROSCOPE CONFIGURATION 140 USING A POINT-SPREAD
FUNCTION TO MEASURE RESOLUTION AND OPTICAL ABERRATIONS 142 MINIMIZING
SPHERICAL ABERRATION BY VARYING THE REFRACTIVE INDEX OF THE IMMERSION
OIL 144 3. IMAGING FLUORESCENT SIGNALS IN FIXED DROSOPHILA EMBRYOS AND *
EGG CHAMBERS 146 DIFFICULTIES IMAGING FLUORESCENT SIGNAL INSIDE A
BIOLOGICAL SPECIMEN 146 MOUNTING FIXED DROSOPHILA EMBRYOS AND EGG
CHAMBERS 147 CHOOSING THE CORRECT MOUNTING MEDIUM 148 4. RE-ASSIGNING OR
REMOVING OUT-OF-FOCUS LIGHT 149 DECONVOLUTION: BASIC PRINCIPLES 150
DECONVOLUTION: PRACTICAL CONSIDERATIONS 151 DECONVOLUTION OF IMAGES FROM
THICK SPECIMENS 152 INCOMPLETE DECONVOLUTION AND DECONVOLUTION ARTEFACTS
152 5. TIME-LAPSE MICROSCOPY OF LIVING DROSOPHILA EMBRYOS AND EGG
CHAMBERS EXPRESSING GFP MARKERS OR MICRO-INJECTED WITH FLUORESCENT
REAGENTS 153 MOUNTING, MICRO-INJECTING AND IMAGING LIVING EMBRYOS 153
FIXING MICRO-INJECTED DROSOPHILA EMBRYOS 155 FIXING GFP-EXPRESSING
EMBRYOS 156 XVII CONTENTS PREPARATION, MOUNTING AND IMAGING LIVING EGG
CHAMBERS 157 CHOICE OF EXPRESSION VECTORS 158 CHOICE OF GFP MUTANTS 158
CHOICE OF FILTER CUBES * 160 6. TROUBLESHOOTING COMMON IMAGING PROBLEMS
160 ACKNOWLEDGEMENTS R , 161 REFERENCES 161 7. GREEN FLUORESCENT
PROTEIN IN PLANTS 163 CHRIS HAWES, PETRA BOEVINK AND IAN MOORE 1.
INTRODUCTION , 163 ,2. CHOICE OF GFP , 163 3. EXPRESSION SYSTEMS 165
VIRUS-MEDIATED EXPRESSION OF GFP 165 TRANSIENT EXPRESSION 169
AGMBQCTERIUM-MEDIAT&D TRANSIENT EXPRESSION IN NICOTIANA SP. 171 4.
DOUBLE LABELLING 174 5. MICROSCOPY 175 REFERENCES 176 8. FLUORESCENCE
MICROSCOPY IN YEAST 179 IAIN M. HAGAN AND KATHRYN R. AYSCOUGH 1.
INTRODUCTION . 179 2. PRACTICAL CONSIDERATIONS WHEN WORKING WITH YEAST
181 DIFFICULTIES ENCOUNTERED WHEN WORKING WITH YEAST 181 GROWING YEAST
181 HARVESTING AND FIXING CELLS 182 CONSIDERATIONS WHEN FIXING YEAST
CELLS 182 CHEMICAL FIXATION TO CROSS-LINK PROTEINS 182 QUENCHING 183 THE
DURATION OF CHEMICAL FIXATION IS A KEY CONSIDERATION 183 FIXING BY
PROTEIN PRECIPITATION: SOLVENT FIXATION 183 CONSIDERATIONSWHEN
HARVESTING CELLS 184 CELL WALL DIGESTION 185 ANTIBODIES , 185 PRIMARY
ANTIBODIES 185 SECONDARY ANTIBODIES 188 DOUBLE LABELLING 188 EPITOPE
MASKING 189 XVIII CONTENTS MOUNTING CELLS IMMUNOFLUORESCENCE MICROSCOPY
CELLS EXPRESSING GFP-TAGGED PROTEINS VISUALIZING CELLS CHROMATIN
STAINING 3. MOLECULAR MANIPULATIONS USING MOLECULAR TAGS TO VISUALIZE
PROTEINS FUNCTIONALITY EXPRESSING PROTEIN TAGS 4. LOCALIZING A NOVEL
PROTEIN IN S. CEREVISIAE 5. HOW TO LOCALIZE A NOVEL PROTEIN IN S. POMBE
6. LOCALIZING PROTEINS AND ORGANELLES BY OTHER FLUORESCENCE METHODS
ACKNOWLEDGEMENTS REFERENCES APPENDICES AL: EXCITATION AND EMISSION
MAXIMA OF COMMONLY USED FLUOROPHORES A2: COMMERCIAL SOURCES OF
ANTIBODIES A3: OPTICAL UNITS A4: LIST OF SUPPLIERS GLOSSARY OF
MICROSCOPY TERMS INDEX 189 189 191 191 191 192 192 193 194 194 197 201
205 205 207 209 212 213 223 225 XIX
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| indexdate | 2024-07-09T18:38:08Z |
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| isbn | 0199637407 0199637415 |
| language | English |
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| physical | XXXIV, 231 S. Ill., graph. Darst. |
| publishDate | 2000 |
| publishDateSearch | 2000 |
| publishDateSort | 2000 |
| publisher | Oxford Univ. Press |
| record_format | marc |
| series | The practical approach series |
| series2 | The practical approach series |
| spelling | Protein localization by fluorescence microscopy a practical approach ed. by Victoria J. Allan Oxford [u.a.] Oxford Univ. Press 2000 XXXIV, 231 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 218 Fluorescence microscopy Green fluorescent protein Immunofluorescence Proteins Analysis Proteine (DE-588)4076388-2 gnd rswk-swf Fluoreszenzmikroskopie (DE-588)4290958-2 gnd rswk-swf Fluoreszenzmikroskopie (DE-588)4290958-2 s Proteine (DE-588)4076388-2 s DE-604 Allan, Victoria J. Sonstige oth The practical approach series 218 (DE-604)BV007921321 218 HEBIS Datenaustausch Darmstadt application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008885513&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Protein localization by fluorescence microscopy a practical approach The practical approach series Fluorescence microscopy Green fluorescent protein Immunofluorescence Proteins Analysis Proteine (DE-588)4076388-2 gnd Fluoreszenzmikroskopie (DE-588)4290958-2 gnd |
| subject_GND | (DE-588)4076388-2 (DE-588)4290958-2 |
| title | Protein localization by fluorescence microscopy a practical approach |
| title_auth | Protein localization by fluorescence microscopy a practical approach |
| title_exact_search | Protein localization by fluorescence microscopy a practical approach |
| title_full | Protein localization by fluorescence microscopy a practical approach ed. by Victoria J. Allan |
| title_fullStr | Protein localization by fluorescence microscopy a practical approach ed. by Victoria J. Allan |
| title_full_unstemmed | Protein localization by fluorescence microscopy a practical approach ed. by Victoria J. Allan |
| title_short | Protein localization by fluorescence microscopy |
| title_sort | protein localization by fluorescence microscopy a practical approach |
| title_sub | a practical approach |
| topic | Fluorescence microscopy Green fluorescent protein Immunofluorescence Proteins Analysis Proteine (DE-588)4076388-2 gnd Fluoreszenzmikroskopie (DE-588)4290958-2 gnd |
| topic_facet | Fluorescence microscopy Green fluorescent protein Immunofluorescence Proteins Analysis Proteine Fluoreszenzmikroskopie |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008885513&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV007921321 |
| work_keys_str_mv | AT allanvictoriaj proteinlocalizationbyfluorescencemicroscopyapracticalapproach |