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Laboratory Equipment
LSR II
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BD LSR II - Chapter 5: Calcium Flux Analysis
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87
5
Calcium Flux
The following topics are
covered in t
his chapter:
•
In
tracellular Calcium Co
ncentration on p
age
88
•
Calcium Flux Optimization on
page
89
•
Recording Calcium Flux Data on page
96
LSR2
.book
Pag
e
87 Tu
es
day, Apri
l 25
, 200
6 3:
34 PM
86
88
Table of Contents
Main Page
Default Chapter
5
Table of Contents
5
About this Guide
11
Conventions
11
BD LSR II Documentation
12
BD LSR II Online Help
12
Printed Documentation
13
Electronic Documentation
14
Technical Assistance
15
Safety and Limitations
17
Laser Safety
17
Laser Product Classification
18
Precautions for Safe Operation
18
Electrical Safety
19
Biological Safety
20
General Safety
21
Symbols and Labels
21
Limitations
23
Chapter 1: Introduction
25
Overview
26
Components
26
Power Switch
26
Control Panel
27
Handles
27
Fluidics
28
Sample Flow Rate Control
28
Fluid Control
28
Sample Injection Port
29
Sheath and Waste Containers
31
Optics
32
Lasers
33
Filters
34
Detectors
35
BD LSR II Workstation
36
Chapter 2: Instrument Setup
37
Starting the Cytometer and Computer
38
Setting up the Optical Filters and Mirrors
39
Filter and Mirror Configurations
40
Changing Optical Filters or Mirrors
42
Filter and Mirror Specifications
43
Preparing Sheath and Waste Containers
44
Preparing the Sheath Container
44
Preparing the Waste Container
46
Preparing the Fluidics
47
Removing Air Bubbles
47
Priming the Fluidics
49
Quality Control
49
Chapter 3: Running Samples
51
Sample Optimization Using Instrument Setup
52
Verifying Instrument Configuration and User Preferences
54
Creating an Experiment
56
Adjusting the Voltages and Threshold
59
Calculating Compensation
63
Recording and Analyzing Data
65
Preparing the Workspace
65
Recording Data
67
Analyzing Data
69
Reusing the Analysis
73
Saving the Analysis
73
Chapter 4: DNA Analysis
75
Criteria for DNA Experiments
76
DNA Setup
77
How to Use DAPI with DNA QC
77
How to Use PI with DNA QC
78
CEN Optimization
78
Preparing the Workspace
78
Running CEN
82
CTN Resolution
84
Running CTN
84
Optimization for Data Recording
86
Chapter 5: Calcium Flux
87
Intracellular Calcium Concentration
88
Calcium Flux Optimization
89
Using the Time Parameter
89
Setting up the Experiment
90
Optimizing for Calcium Flux
94
Recording Calcium Flux Data
96
Chapter 6: Maintenance
99
Daily Cleaning and Shutdown
100
Daily Fluidics Cleaning
100
Daily Shutdown
102
Scheduled Maintenance
103
System Flush
103
Waste Management System Maintenance
105
Periodic Maintenance
107
Changing the Sheath Filter
108
Changing the Bal Seal
110
Changing the Sample Tube O-Ring
112
Chapter 7: Troubleshooting
113
Instrument Troubleshooting
114
Appendix A: Technical Overview
123
Fluidics
124
Optics
125
Light Scatter
125
Fluorescence
126
Optical Filters
127
Compensation Theory
131
Electronics
134
Pulse Measurements
135
Digital Electronics
136
BD Facsdiva Option White Paper
136
Threshold
137
Laser Controls
137
Appendix B: BD LSR II QC Log
139
Appendix C: Filter Templates
143
Octagon Template
145
Trigon Template
147
Appendix D: Supplies and Consumables
149
QC Particles
150
Reagents
151
Equipment
152
Appendix E: Standard Default Configuration
153
Standard (4-Blue 2-Violet 2-UV 2-Red) Configuration Specification
154
Default Configuration
155
Octagon and Trigon Maps
156
Custom Configurations
159
Appendix F: Special Order Configurations
165
Common Special Order Configurations
166
6-Blue 0-Violet 0-UV 3-Red Configuration
167
6-Blue 2-Violet 0-UV 3-Red Configuration
168
6-Blue 0-Violet 2-UV 3-Red Configuration
169
6-Blue 2-Violet 2-UV 3-Red Configuration
171
6-Blue 6-Violet 0-UV 3-Red Configuration
172
6-Blue 6-Violet 0-UV 4-Red Configuration
174
6-Blue 6-Violet 2-UV 3-Red Configuration
176
6-Blue 6-Violet 2-UV 4-Red Configuration
178
Special Order Configuration Trigon and Octagon Maps
180
6-Color Blue Octagon Default Configuration Map
181
2-Color Violet Trigon Default Configuration Map
182
6-Color Violet Octagon Default Configuration Map
183
2-Color UV Trigon Default Configuration Map
184
3-Color Red Trigon Default Configuration Map
185
4-Color Red Octagon Default Configuration Map
186
BD FACS Instrument Configuration Application
187
Installing ICA
187
Updating Your Default Configuration
188
Appendix G: Setting Laser Delay
193
About Laser Delay
194
Optimizing Laser Delay
195
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