Page 3 - Live-cellanalysis handbook

Table of Contents

1. Introduction ..........................................................................................................................................................................3

2. From Images to Answers ...................................................................................................................................................5
3. Cell Culture Quality Control Assays ..............................................................................................................................9

4. Kinetic Cell Health and Viability Assays ...................................................................................................................13
a. Proliferation .....................................................................................................................................................................16
b. Caspase-3/7 and Annexin Apoptosis ..........................................................................................................................28
c. Cytotoxicity ......................................................................................................................................................................36
5. Kinetic Cell Migration and Invasion Assays .............................................................................................................44
a. Scratch Wound ................................................................................................................................................................47
b. Chemotaxis .......................................................................................................................................................................52
c. Transendothelial Migration ..........................................................................................................................................58

6. Kinetic Assays for Quantifying Protein Dynamics .................................................................................................63
a. Antibody Internalization ...............................................................................................................................................65
b. Live-Cell Immunocytochemistry .................................................................................................................................72

7. Kinetic Assays Utilizing Complex Models ..................................................................................................................80
a. Multi-Spheroid ................................................................................................................................................................83
b. Single Spheroid ...............................................................................................................................................................90

8. Kinetic Assays for Studying Neuronal Models ........................................................................................................99
a. Neurite Analysis ........................................................................................................................................................... 101
b. Neuronal Activity ........................................................................................................................................................ 109
c. Neuroimmune Assays .................................................................................................................................................. 118
9. Appendix ........................................................................................................................................................................... 127
a. Cell Health Protocols
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i. IncuCyte Cell Proliferation Assay ................................................................................................................. 128
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ii. IncuCyte Transduction ..................................................................................................................................... 130
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iii. IncuCyte Apoptosis Assay ............................................................................................................................... 132
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iv. IncuCyte Cytotoxicity Assay ........................................................................................................................... 135
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v. IncuCyte Immune Cell Killing Assay ............................................................................................................. 138
b. Cell Migration and Invasion Protocols
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i. IncuCyte Scratch Wound Assay ..................................................................................................................... 142
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ii. IncuCyte Chemotaxis Cell Migration Assay ................................................................................................ 147
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iii. IncuCyte Chemotaxis Cell Invasion Assay ................................................................................................... 151
iv. Chemotactic Transendothelial Migration Assay .......................................................................................... 154
c. Protein Dynamics Protocols
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i. IncuCyte Antibody Internalization Assay ..................................................................................................... 157
ii. IncuCyte Live-cell Immunocytochemistry .................................................................................................... 160
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d. Spheroid Protocols
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i. IncuCyte Multi-Spheroid Assay ...................................................................................................................... 165
ii. IncuCyte Single Spheroid Viability Assay .................................................................................................... 168
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e. Neuroscience Protocols
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i. IncuCyte Label-Free Neurite Outgrowth and Cell Health Analysis Assay ............................................ 170
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ii. IncuCyte Fluorescence Neurite Analysis Assay ........................................................................................... 174
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iii. IncuCyte Neuronal Activity Assay ................................................................................................................. 180
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iv. IncuCyte pHrodo Orange Phagocytosis Assay ........................................................................................... 187
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