Transcription
CyAn ADP with Summit SoftwareUser GuideDocument Number 0000050Revision FJuly 2006
Copyright 2001 - 2006 Dako. All rights reserved.This document may not be copied in whole or in part or reproduced in any other media withoutthe express written permission of Dako. Please note that under copyright law, copying includestranslation into another language.2CyAn ADP with Summit Software User Guide
The CyAn ADP Analyzer is the next step in Advanced Digital Processing (ADP) with enhanceddetection for flow cytometry. The CyAn ADP High-Performance Flow Cytometer is a research toolengineered for precision analysis of cells, bacteria, and other similarly sized particles. With CyAnADP, Dako sets a new industry standard with a combination of features never before available ona bench-top analyzer. CyAn ADP gives users three excitation lines with independent, alignmentfree focusing optics, simultaneous nine color and two scatter parameters, analysis rates of 70,000events per second, a full 9 9 interlaser compensation matrix, and high sensitivity. The result isstable, user-friendly, and flexible technology. The instrument is optimized for cell cycle, kinetics,fluorescent protein work, and multi-color immunophenotyping. Rare-event analysis, such as MHCDextramer studies, and no-lyse whole blood applications are easily performed on the CyAn ADP.The instrument also provides simplified compensation before, during, and after acquisition withunequaled sensitivity in all fluorescent parameters.CyAn ADP with Summit Software User Guide3
Contacting DakoDako should be contacted immediately for assistance in the event of any instrument malfunction.For further information please contact your local Dako office.AustraliaTel. 2 9316 4633Fax 2 9316 4773GermanyTel. 040 69 69 470Fax 040 69 52 741SwitzerlandTel. 041 760 11 66Fax 041 760 11 77AustriaTel. 0800 0800 7153Fax 0800 0800 7154ItalyTel. 02 58 078 1Fax 02 58 078 292United KingdomTel. (0)1 353 66 99 11Fax (0)1 353 66 89 89Technical SupportTel. (0) 1 353 66 99 65BelgiumTel. 016 38 72 20Fax 016 38 72 21JapanTel. 075 211 3655Fax 075 211 1755United States of AmericaCarpinteria, CaliforniaTel. 805 566 6655Fax 805 566 6688CanadaTel. 905 858 8510Fax 905 858 8801The NetherlandsTel. 020 42 11 100Fax 020 42 11 101Czech RepublicTel. 420 541 423 710Fax 420 541 423 711NorwayTel. 23 14 05 40Fax 23 14 05 42DenmarkHead OfficeTel. 44 85 95 00Fax 44 85 95 95PolandTel. 058-661 1879Fax 058-661 3390SalesTel. 44 85 97 56Fax 44 85 84 29SpainTel. 93 499 05 06Fax 93 499 02 08FranceTel. 1 30 50 00 50Fax 1 30 50 00 11SwedenTel. 08 556 20 600Fax 08 556 20 6194Technical SupportTel. 800 424 0021Customer ServiceTel. 800 235 5763United States of AmericaFort Collins, ColoradoFlow InstrumentationTel. 800 822 9902Fax 970 226 0107CyAn ADP with Summit Software User Guide
User ResourcesFor the latest information on Dako products and services, please visit the Dako Web site athttp://www.dako.com, and the Dako Flow Cytometry Discussion Group athttp://www.dakousa.com/index/support/discussion groups.htm.ScopeThis guide provides a detailed discussion of the architecture and operating procedures for the CyAn ADP High-Performance Flow Cytometer. Detailed operating instructions for Summitsoftware can be found in the Summit software online help system. The information contained inthis document can be applied to all CyAn ADP products.DisclaimersThis document is not a substitute for the detailed operator training provided by Dako or for otheradvanced instruction in general cytometric techniques. It is essential that the operator have a working knowledge of Microsoft Windows XP or current Operating System prior to using thisguide. Dako also recommends the use of all of the networking services provided in the individualoperator’s laboratory. Although daily maintenance and routine instrument adjustments arediscussed here, your local Dako Technical Service Group should be contacted immediately forassistance in the event of any instrument malfunction.Trademarks Microsoft and Windows XP are registered trademarks of Microsoft Corporation. Macintosh is a registered trademark of Apple, Inc. MoFlo is a registered trademark of Dako. CyAn ,Summit , SpectrAlign , and SpectraComp are trademarks of Dako. All other trade namesand trademarks are the property of their respective holders.CyAn ADP with Summit Software User Guide5
Table of ContentsSection 1 Safety . 9General Hazard Avoidance . 10Biohazard Avoidance . 11Laser Safety . 12Section 2 Installation. 15CyAn ADP Installation Requirements . 15General Laboratory Information. 15CyAn ADP Power Configuration and Consumption . 17CyAn ADP Instrument Configurations. 17Section 3 System Overview. 19CyAn ADP Features . 19CyAn ADP Subsystems . 20Fluidics . 21Optics . 22Electronics . 25Peripheral Devices. 26Software . 26Summit Software Features . 27Section 4 Startup and Shutdown Procedures . 29Required Reagents for Startup and Shutdown . 29CyAn ADP Startup. 29Daily Decontamination Procedure. 32Shutdown Procedure. 33Section 5 Sheath Management System. 35Sheath Management System Indicators . 35Changing Out Sheath and Waste Containers. 41Replacing Cleaner Fluid. 42Section 6 Summit Software Basics . 43Acquire and Analyze up to 10 Million Events. 43Create a New Database. 44Open an Existing Database . 46Create a Histogram . 47Create a Dot Plot. 48Open FCS (Listmode) Files. 49Set a Gate from a Single Region . 51Set a Serial Gate . 52Define Gate Logic. 55Shortcut Keys . 57Section 7 CyAn ADP Maintenance. 59Daily Preventive Maintenance. 59Weekly Preventive Maintenance. 59Reusable Sheath Containers . 59Head Unit and SMS Cart . 60Fluidic Lines . 60Monthly Preventive Maintenance . 60System Decontamination . 60Laser Interlock Maintenance Procedure . 61Biannual Maintenance Procedure . 61Section 8 Troubleshooting . 63Section 9 Six-Color Compensation Experiment. 65Background . 65Setting up for sample acquisition . 66Setting Up Histograms for Compensation. 686CyAn ADP with Summit Software User Guide
Manual Compensation . 71Setting Spillover Percentage for Compensation . 72Running the Second Sample . 73Running the Remaining Samples. 75Running the “All Stains” Sample . 83Putting Compensation into Practice . 84CD3 T-Lymphocytes . 84CD 19 B-Lymphocytes . 84CD4 /CD3 T-Lymphocytes (T-Helper cells) . 85CD8 /CD3 T-Lymphocytes (T-Cytotoxic/Suppressor cells). 85Natural Killer Lymphocytes . 86Section 10 Auto Compensation . 87Auto Compensation for Single Positive Controls . 87Section 11 VisiComp . 91Using the VisiComp Feature after Compensation. 91Section 12 WorkList Builder. 93Overview . 93Accessing the WorkList Builder. 94Creating a New Panel . 95Setting up Tests and Defining Reagents for a Panel . 97Creating Reagents by Defining Antibody/Fluorochrome Combinations. 98Deleting Reagents from the WorkList Builder Database. 100Setting up Tests (Tubes or Samples) within a Panel . 101Setting up a Panel . 105Creating Worklists from Defined Panels and Tubes . 107Setting up Worklists for Multiple Experiments or Specimens. 108Customizing the WorkList Builder Viewing Area. 110Edit Columns. 110Importing a Specimen List from a Text File . 111Saving and Reusing Worklists . 112Associating Protocols, Gate/Event Limits, or Other Information to Worklist Tests . 113Appendix A Approved Cleaners and Disinfectants . 115Cleaners . 115Disinfectants for Use in Sample Line*. 115Disinfectants for Use in Sheath Line** . 115Disinfectants for Use in the Waste Tank . 115Appendix B CyAn ADP Consumables and Parts . 117Appendix C Technical & Instrument Specifications . 119Appendix D Symbol Definitions . 123Appendix E Flow Cytometry References . 125CyAn ADP with Summit Software User Guide7
8CyAn ADP with Summit Software User Guide
Section 1SafetyElectrical SafetyThe Dako instrument product line conforms to international regulationsencompassing the accessibility of high voltages by the user. In the United States,each flow cytometer is manufactured to comply with applicable regulations from theAmerican National Standards Institute (ANSI), the Occupational Safety and HealthAdministration (OSHA), and various state regulatory organizations. Internationally,each CyAn ADP High-Performance Flow Cytometer is manufactured to complywith the International Electrotechnical Commission’s Standard for SafetyRequirements for Electrical Equipment for Measurement, Control and LaboratoryUse (IEC 61010-1) and subsequent amendments.Biohazard SafetyThe CyAn ADP instrument may be used for the analysis of pathogenic or otherharmful agents. These operations can constitute a biohazard for the operator as wellas bystanders in the surrounding laboratory space. Dako does not certify anyinstrumentation for use with any hazardous organism or agent. Dako stronglyrecommends explicit guidance be obtained from the institutional or companybiosafety officer prior to operating this instrument with any potentially hazardousorganism or agent. See the Biohazard Avoidance section for more information.WARNINGIf any biohazardous organism or agent is used in this instrument, the user mustinform Dako in writing prior to any field service visit or the return of any part to Dako,or its vendors, for service. Safety of the CyAn ADP user and of all Dako employeesis of primary concern. Proper decontamination procedures must be followed forreturned parts.WARNINGWear Personal Protective Equipment (PPE) in accordance with your laboratorysafety procedures when operating or maintaining this instrument. It is recommendedthat gloves, a laboratory coat, and safety glasses are worn at all times when usingthe CyAn ADP. Use of this instrument in a manner other than that specified in thismanual may cause impairment of equipment, compromise safety, and/or result ininjury.CyAn ADP with Summit Software User Guide9
General Hazard AvoidanceSafety of the CyAn ADP operator is of primary concern. It is important that all operatorsfamiliarize themselves with this safety section to avoid all potential hazards associated with theuse of the CyAn ADP. Any questions or concerns regarding safety of the instrument should bedirected to Dako Colorado immediately.1. PINCHING: The lid of the CyAn ADP is on hinges that should keep the lid in the UP positionwhen open. It is recommended by Dako that the lid NOT be left open for extended periods oftime OR be opened and closed repeatedly to avoid excessive wear on the hinges which may,over time, cause the lid to fall unexpectedly, potentially falling on an operator’s hands orfingers, causing injury. Please use caution when the lid is open and do not leave unattended.If the lid of the CyAn ADP becomes loose and will not stay up on its own, please contactCustomer Care so it can be repaired.2. TRIPPING: The Sheath Management System (SMS) is attached to the CyAn ADP head unitby an 8 foot (2.4 meters)long umbilical cord. It is suggested that the SMS cart be placedunderneath a laboratory bench to keep it out of the way of foot traffic through the laboratory.If this is not possible, it is recommended that the SMS be placed where it will minimize thelikelihood of a tripping hazard. It is suggested that the umbilical cord be run behind thelaboratory bench, if possible, to keep the cord from dangling and potentially creating atripping hazard as well. If the cord must be run down in front of the laborat
The CyAn ADP High-Performance Flow Cytometer is a research tool engineered for precision analysis of cells, bacteria, and other similarly sized particles. With CyAn ADP, Dako sets a new industry standard with a combination of features never before available on a bench-top analyzer. CyAn ADP gives users three excitation lines with independent .
