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FLIPR Tetra
Molecular Devices FLIPR Tetra User Manual
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Running an Experiment
196
0112-0109 H
195
197
Table of Contents
Default Chapter
3
Table of Contents
3
Chapter 1: System Overview
13
Fluorescence Mode
13
Luminescence Mode
14
System Requirements
14
Electrical
14
Minimum Space
15
Chapter 2: System Hardware Features
17
Overview of FLIPR Tetra System Hardware Features
17
System Diagram
18
Plate-Handling System
18
Five-Position Stage
18
Plates
21
Instrument Status Panel
21
Manual Mode
22
Robotic Integration
23
Observation Panel
24
Liquid-Handling System
25
Standard Pipettor Head
25
Cell Suspension
27
Pin Tool Head
29
Compatible Plate Configurations
30
Tip and Pin Tool Loading
31
Tip and Pin Tool Washing
31
Optical System
32
CCD Camera Options
33
LED Modules
35
Emission Filters
37
Chiller
38
Computer System
38
Host Computer
38
Embedded Computer
39
Chapter 3: Startup and Shutdown
41
Starting up the System
41
Shutting down the System
42
Chapter 4: Software Installation
43
Installing Screenworks Software
43
Activating the Screenworks Peak Pro License
44
Online Vs. Offline Installation
45
Online (Instrument) Mode
45
Offline (Desktop) Mode
46
Uninstalling Screenworks Software
46
Chapter 5: Screenworks Software Overview
47
Screenworks Software Main Screen
47
Title Bar
48
Menu Bar
48
Toolbar
48
Status Bar
51
Menu Bar
51
File Menu
52
View Menu
54
Instrument Menu
55
Tools Menu
61
Window Menu
65
Help Menu
65
Instrument Status
66
Instrument Status Tab
66
Instrument Configuration Tab
68
Process Explorer Tab
69
Experiment Window
69
Constructing Protocols Using FLIPR Tetra Processes
70
Settings Process
71
Setup Read Mode
72
Assign Plate to Position
74
Data File Name
76
Folders
76
Temperature Control
77
Auto Print Options
78
Analysis Process
79
Viewing Data
79
Analyzing Data
87
Exporting Data
99
Batch Export
106
Image Display
107
Notes
107
Transfer Fluid Process
108
Aspirate Table (Standard Pipettor)
109
Dispense Table (Standard Pipettor)
112
Aspirate Configuration (Pin Tool)
115
Dispense Configuration (Pin Tool)
116
MIX Fluid Process
117
MIX Fluid (Standard Pipettor)
117
MIX Fluid (Pin Tool)
119
MIX with TF
120
Read Process
120
Read
120
Read with TF
122
Wash Tips or Pins Process
123
Wash Tips (Standard Pipettor)
124
Wash Pins (Pin Tool)
125
Blot Pins Process
126
Pause Pipettor Process
126
Wash Cell Reservoir Process
127
Finish with Source
128
Chapter 6: Exchanging Hardware
129
Exchanging Pipettor and Pin Tool Heads
129
Uninstalling a Pipettor or Pin Tool Head
129
Installing the Pipettor Head
131
Uninstalling Wash Reservoir Top
133
Installing Wash Reservoir Top
134
Resetting FLIPR Tetra System after Changing Pipettor Heads
134
Exchanging the 1536 Tip Gasket
135
Installing the Gasket
135
Removing the Tip Block and Gasket
136
Exchanging Pin Tools
137
Loading and Unloading the Pin Tool
137
Exchanging LED Modules
138
Uninstalling LED Modules
138
Installing LED Modules
139
Changing Filters
141
Uninstalling a Filter
142
Installing an Emission Filter
143
Installation of Plate Hold-Down Devices
144
Selection of Appropriate Plate Hold-Down
144
Cell Reservoir
146
Installing Cell Reservoir
146
Uninstalling Cell Reservoir
147
Chapter 7: Calibration and Signal Test
149
Optical Calibration
149
Adding a Read Plate to Plate Library
149
Recalibrating the Optics
150
Running a Signal Test
153
System Check
153
Running a Plate Prior to an Experiment
154
Chapter 8: Running an Experiment
157
Overview
157
Preparing Cells for Adherent Assays
157
Location of Cells in the Plate
157
Cell Densities
158
Cell Seeding
158
Preparing Cells for Suspension Assays
159
Location of Cells in the Plate
159
Cell Densities
159
Cell Seeding
159
Powering-Up the System
159
Checking the System
160
Running the Yellow Plate Signal Test
160
Dye-Loading the Cells for Fluorescence Assays
161
Loading Duration and Temperature
161
Preparing a Source/Compound Plate
162
Preparation Time for the Source Plate
162
Recommended Source Plates
162
Concentration of Compounds in the Source Plate
162
Addition and Mixing of Compounds to the Cell Plate
162
Compound Plates for Suspension Assays
163
Setting up an Assay Protocol
164
Creating a Protocol File
164
Optimizing the Optics and Fluid Dispensing
165
Optimizing the Hardware Settings
165
Optimizing Optics Hardware
165
Adjusting the Pipettor Height
167
Adjusting the Fluid Dispensing Speed
168
Optimizing Fluid Volume
168
Optimizing Pin Tool Delivery
169
Optimizing Cell Delivery
169
Start the Assay Run
169
FLIPR Calcium Assay Kit Protocol
170
Required Materials
170
Calcium Assay Kit Experimental Protocol
172
Running the FLIPR Calcium Assay Kit
174
Troubleshooting Guide
176
FLIPR Membrane Potential Assay Kit Protocol
177
Required Materials
177
Cell Preparation for the FLIPR Membrane Potential Assay179
179
Dye Loading Using the FLIPR Membrane Potential Assay Kit
180
Running the FLIPR Membrane Potential Assay
181
Troubleshooting the FLIPR Membrane Potential Assay Kit
183
Voltage Sensor Probes Assay Protocol
184
Required Materials
184
Cell Preparation for the Voltage Sensor Probe Assay
185
Running the Voltage Sensor Probe Assay
187
Luminescence Assay Protocol
188
Required Materials
188
Coelenterazine Loading for Adherent Assays
189
Coelenterazine Loading for Suspension Cell Assays
190
Preparation of Cell Reservoir and Running the Assay
191
Optimizing an Assay
194
Chapter 9: Troubleshooting
197
Instrument Status Colors
197
Troubleshooting Start-Up
198
General Troubleshooting
200
Troubleshooting the Pipettor
205
Troubleshooting the Optics
210
Troubleshooting the Yellow Plate
215
Troubleshooting the Tip Washer
216
Troubleshooting the Cell Reservoir
218
Troubleshooting Data
218
Troubleshooting Robotic Integration
219
Appendix A: Robotic Integration
221
Introduction
221
Purpose
221
Document Conventions
221
Interface Versioning
221
Instrument Overview
222
Instrument Basic Function
222
Instrument Hardware Introduction
222
Instrument Layout
224
Plate Layout
225
Plate-Handling System
225
Instrument Layout Terminology (I.e., Where's the Front?)
226
Robotic Plate Loading
226
Optics Access Door
227
Washer Placement
227
Cell Suspension Placement
228
Other Instrument Access Areas
228
Monitor and Keyboard Placement
230
Required Access Areas
230
FLIPR Tetra System Control Architecture
233
General Description
233
Manual Mode
234
Remote Mode
234
Communication Protocol and Address
236
General Remote Mode Use
236
Setting up Protocols for Remote Control Use
240
General Directions
240
Settings Screen
240
Analysis Screen
242
Transfer Fluid Processes
243
Wash Tips Processes
244
MIX Fluid Processes
244
Read Processes
244
Finish with Source Processes
244
Wash Cell Reservoir Processes
244
Command Set
244
Command Syntax
244
Version<Cr
245
Status<Cr
246
Loadplate<Tab>Location, Last Plate[, BAR CODE] <CR
248
Removeplate<Tab>Location <CR
250
Openprotocol<Tab>File Name<Cr
251
Findprotocols<Tab>Folder<Cr
251
Runexperiment<Tab>[Data File Name]<Cr
252
Stopexperiment<Cr
253
Clearerror<Cr
254
Loadtips<Cr
255
Unloadtips<Cr
256
Cyclecameratemp<Cr
257
Tempcontrolonoff<Tab>Temp<Cr
257
Washtips<Tab>Fluid Type, Wash Cycles, Volume/Stroke, Aspirate Speed, Pump Speed, Strokes, Hold Time, Dispense Speed<Cr
258
Configuration <CR
260
Statusex<Cr
262
Cellflaskcontrol<Tab>Rate<Cr
264
Washcellreservoir<Tab>Fluid Type, Fill Speed, Drain Destination, Drain Speed, Wash Cycles, Hold Time, Volume Level<Cr
265
Error Handling and Reporting
267
Error Types
267
Appendix A-Remote Interface Revision History
270
V1.0
270
V1.1
270
V1.2
270
V1.3
271
Appendix B-Bar Code Specifications
271
About Bar Codes
271
Bar Code Recommendations
271
Bar Code Specifications
272
Appendix B: Data Processing Algorithms
273
Hypothetical Experiment
273
Crosstalk Correction
274
Determining Crosstalk Correction
274
Spatial Uniformity Correction
275
Determining Spatial Uniformity Correction
275
Negative Control Correction
276
Determining Negative Control Correction
277
Positive Control Scaling
278
Determining the Positive Control Scaling
278
Subtract Bias
279
Determining Subtract Bias
279
Response over Baseline
280
Determining Response over Baseline Correction
280
Appendix C: Consumables and Accessories
283
FLIPR Tetra System Accessories
283
Field Installations
283
Pipettor Heads
283
Optics Consumables
284
Pipetting Consumables
285
Cell Reservoir Consumables
286
Plates
287
96-Well Read Plates
287
96-Well Read Plate Masks
287
96-Well Source Plates
288
384-Well Read Plates
289
384-Well Source Plates
290
1536-Well Read Plates
290
1536-Well Source Plates
291
Source Reservoirs
291
Assays Performed on the FLIPR Tetra System
291
Calcium Flux Consumables
291
FLIPR Membrane Potential Assay Kit Consumables
293
Equipment and Supplies Suggested to Perform Assays with Your FLIPR Tetra System
293
Appendix D: Using Aquamax Sterilant
295
Principle of Use
295
Contents
295
Materials Required but Not Provided
295
Storage
296
Reagent Preparation
296
Reagent Use
296
Warnings, Precautions and Limitations
296
Appendix E: Decontamination Certificate
299
Procedure for Completing the Certificate
299
Appendix F: Electromagnetic Compatibility (EMC)
301
Regulatory Information for Canada (Ices/Nmb-001:2006)
301
ISM EQUIPMENT CLASSIFICATION (Group 1, Class A)
301
Information for the User (Fcc Notice)
301
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Molecular Devices FLIPR Tetra Specifications
General
Brand
Molecular Devices
Model
FLIPR Tetra
Category
Laboratory Equipment
Language
English
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