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Appendix E: QC
Using BD FAC
SDiva Software
321
Figure E-4
F
SC area scaling factor set incorrectly (left) and correctly (
right)
1
2
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BD FACSAria II
1
User’s Guide
1
FCC Information
3
Contents
5
About This Guide
13
Conventions
14
Technical Assistance
15
Limitations
16
Cytometer Components
17
Fluidics Cart
18
Power and Operation
21
Flow Cytometer
22
Fluidics Components
23
Sample Injection Chamber
25
Tube Holders
26
Cuvette Flow Cell
27
Figure 1-11 Nozzle
28
Sort Block
29
Deflection Plates
30
Aspirator Drawer
30
Aerosol Management
31
Sort Collection Chamber
32
Optics System
33
Collection Optics
35
Detectors
36
Stream-Viewing Optics
38
Cytometer Electronics
39
Emergency Stop Button
40
Workstation
41
Theory of Operation
43
Fluid Movement
44
Sample Flow
46
Hydrodynamic Focusing
47
Signal Generation
48
Fluorescent Signals
49
Signal Detection
50
Longpass Filters
52
Bandpass Filters
53
% transmission
54
BP 500/50 filter
54
DF 500/50 filter
54
Neutral Density Filters
55
Electronic Processing
57
Pulse Parameters
58
Laser Delay
59
Drop Formation
61
Breakoff Window
62
Side Stream Formation
65
Drop Delay Overview
67
Drop Charging
68
Yield Mask
70
Purity Mask
71
Phase Mask
72
Sort Precision Modes
73
Defining New Precision Modes
75
Using BD FACSDiva Software
77
Workspace Components
78
Cytometer Controls
79
Change Sample Filter
80
Cleaning Modes
80
Sheath Pressure
80
Sample Agitation
81
Sample Temperature
82
Fluidics Level Indicators
83
Cytometer Configuration
84
Cytometer Status Report
88
Custom Configurations
90
Copying a Base Configuration
92
Select the Sort Setup that
95
Configuration Mismatch Dialog
96
Acquisition Controls
97
Sorting Controls
98
Sort Menu
100
Sort Setup
101
Sort Layout
103
Setting Up a Sort Layout
105
Editing a Sort Layout
107
Using Sorting Controls
107
Using Counters
108
Monitoring a Sort
109
Sort Report
110
Templates
112
Running Samples
113
Cytometer Startup
114
Performing Fluidics Startup
115
Starting the Stream
118
Setting Up the Breakoff
119
Setting Up the Fluidics Cart
122
Refilling the Sheath Tank
123
ATTENTION
124
Emptying the Waste Container
127
Waste (A)
129
Preparing the CS&T Beads
133
Running a Performance Check
134
Reviewing the Results
134
Application Settings
136
Adjusting Area Scaling
137
Figure 4-14 FSC and SSC Plot
138
Optimizing PMT Voltages
141
Saving Application Settings
143
Data Collection
144
Calculating Compensation
148
Data Recording and Analysis
151
Setting Up the Experiment
152
Recording Data
155
Analyzing Data
156
Performing a Batch Analysis
159
Setting Up for Sorting
162
Setting Up for Bulk Sorting
164
Using Manual Drop Delay
168
Overview of Auto Drop Delay
171
Using Auto Drop Delay
171
Pausing and Resuming a Sort
178
Setting Up the Stream
181
Test Sort button
182
Creating a Custom Device
184
Deleting a Custom Device
186
Shutdown and Maintenance
187
Daily Shutdown
188
Fluidics Shutdown
190
External Cleaning
193
Scheduled Maintenance
194
Sample Line Backflush
195
Prime After Tank Refill
196
Prepare for Aseptic Sort
197
DI water line
198
DI water port
198
Purging the Fluid Filters
200
Purging the Sheath Filter
200
Changing Fluid Filters
202
Changing the Sheath Filter
203
Changing the Sample Lines
204
Figure 6-12 Ferrule tool
207
Pinch Valve End
208
Flow Cell End
209
Changing the Air Filters
211
Unscheduled Maintenance
213
Changing Nozzles
214
Cleaning the Nozzle
216
Cleaning the Camera Windows
225
Upper Camera Window
226
Apply lubricant to outside
229
Using Custom Optical Filters
230
Cleaning the Optical Filters
231
FSC ND filter
232
Set screw
232
Nozzle holder
232
Troubleshooting
233
Troubleshooting the Stream
234
Troubleshooting the Breakoff
238
Sorting Troubleshooting
239
Acquisition Troubleshooting
244
Fluidics Troubleshooting
251
Electronics Troubleshooting
252
Technical Specifications
253
Cytometer Specifications
254
Performance
255
Sort Performance
256
Emission Optics
257
Excitation Optics
257
Detector Arrays
258
Fluidics Cart Specifications
259
Appendix A
261
Supplies and Consumables
261
Cytometer Supplies
262
Accessory Kit
264
Other Replacement Parts
266
Consumables
267
Reagents
268
Appendix B
271
Near UV Laser Option
271
System Laser Configurations
272
Specifications
273
Operation
274
Instrument Setup
278
Experiment Setup Tips
278
Appendix C
281
BD Aerosol Management Option
281
Option Components
282
ULPA Filter
283
Starting Up the Evacuator
284
Maintenance
289
Replacing the ULPA Filter
290
Reset button
293
Replacing the Air Filter
294
Appendix D
301
Temperature Control Option
301
Setting Up the Water Bath
303
Cooling OutCooling In
304
Setting Up the Tube Holder
305
Input tubing
306
Output tubing
306
Setting Up the ACDU Stage
307
Starting Up the Water Bath
309
Appendix E
313
QC Using BD FACSDiva Software
313
Software
314
Preparing QC Particles
316
Setting Up the QC Experiment
317
Recording QC Data
325
Figure E-6 Recorded QC data
327
Reusing the QC Experiment
328
Tracking QC Results
329
Numerics
331
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