BD HTS User's Guide

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Transcript of BD HTS User's Guide

BD High Throughput Sampler Users Guidefor the

BD LSR II BD FACSCanto BD FACSCanto II

bdbiosciences.com Part No. 642224 Rev. A bdbiosciences.com April 2007 Part No. 642224 Rev. A June 2007

BD Biosciences BD Biosciences 2350 Qume Drive 2350 Qume Drive San Jose, CA 95131-1807 San Jose, CA 95131-1807 USA USA Tel (877) 232-8995 Tel (877) 232-8995 Fax (408) 954-2347 Fax (800) 325-9637 [email protected]

Asia Pacific Asia Pacific Tel (65) 6-861-0633 Tel (65) 6-861-0633 Fax (65) 6-860-1590 Fax (65) 6-860-1590

Brazil Brazil Tel (55) 11-5185-9995 Tel (55) 11-5185-9995 Fax (55) 11-5185-9895 Fax (55) 11-5185-9895

Europe Europe Tel (32) 2 400 98 95 Tel (32) 2 400 98 95 Fax (32) 2 401 70 94 Fax (32) 2 401 70 94 [email protected] [email protected]

Japan Japan Nippon Becton Dickinson Company, Ltd. Nippon Becton Dickinson Company, Ltd. Toll Free 0120-8555-90 Toll Free 0120-8555-90 Tel 81-24-593-5405 Tel 81-24-593-5405 Fax 81-24-593-5761 Fax 81-24-593-5761

Canada Canada Toll Free (888) 259-0187 Toll Free (888) 259-0187 Tel (905) 542-8028 Tel (905) 542-8028 Fax (888) 229-9918 Fax (888) 229-9918 [email protected] [email protected] Mexico Mexico Toll Free 01-800-236-2543 Toll Free 01-800-236-2543 Tel (52) 55 5999 8296 Tel (52) 55 5999 8296 Fax (52) 55 5999 8288 Fax (52) 55 5999 8288

2007, Becton, Dickinson and Company. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, stored in retrieval systems, or translated into any language or computer language, in any form or by any means: electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without prior written permission from BD Biosciences. The information in this guide is subject to change without notice. BD Biosciences reserves the right to change its products and services at any time to incorporate the latest technological developments. Although this guide has been prepared with every precaution to ensure accuracy, BD Biosciences assumes no liability for any errors or omissions, nor for any damages resulting from the application or use of this information. BD Biosciences welcomes customer input on corrections and suggestions for improvement. BD FACSDiva software 2007, Becton, Dickinson and Company. This software is the property of Becton, Dickinson and Company. Each sale of a stored unit of this software grants the purchaser a nontransferable, nonexclusive, personal license. This software may not be duplicated, reproduced, or copied in any form or by any means whatsoever, except as otherwise permitted by law. This product includes software developed by the Apache Software Foundation (apache.org). BD, BD logo, and all other trademarks are property of Becton, Dickinson and Company. Adobe and Acrobat are registered trademarks of Adobe Systems Incorporated. Diskeeper is a registered trademark of Executive Software International. Microsoft and Windows are registered trademarks of Microsoft Corporation. Sybase, Adaptive Server Anywhere, and SQL Anywhere are trademarks of Sybase, Inc or its subsidiaries. Cy is a trademark of Amersham Biosciences Corp. Cy dyes are subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and are made and sold under license from Amersham Biosciences Corp. only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from Amersham Biosciences Corp., 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA. All other company and product names might be trademarks of the respective companies with which they are associated.

PatentsPerCP: US 4,876,190 APC-Cy7: US 5,714,386

FCC InformationWARNING: Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the users authority to operate the equipment. NOTICE: This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to Part 15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his or her own expense. Shielded cables must be used with this unit to ensure compliance with the Class A FCC limits. This Class A digital apparatus meets all requirements of the Canadian Interference-Causing Equipment Regulations. Cet appareil numrique de la classe A respecte toutes les exigences du Rglement sur the matriel brouilleur du Canada.

NoticeBD Biosciences delivers software and workstations that are intended for running the cytometers supplied by BD Biosciences. It is the responsibility of the buyer/user to ensure that all added electronic files including software and transport media are virus free. If the workstation is used for Internet access or purposes other than those specified by BD Biosciences, it is the buyer/users responsibility to install and maintain up-to-date virus protection software. BD Biosciences does not make any warranty with respect to the workstation remaining virus free after installation. BD Biosciences is not liable for any claims related to or resulting from buyer/user's failure to install and maintain virus protection.

HistoryRevision 338642 640756 Date 10/04 3/06 Change Made Initial release Updated to include BD FACSDiva software, version 5.0, and compatibility with BD FACSCanto and BD FACSCanto II flow cytometers. Updated to include BD FACSDiva software, version 6.0; added configuration and performance tracking module; operational improvements; and increased cytometerspecific improvements.

642224

6/07

ContentsAbout This Guide Conventions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Technical Assistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chapter 1: Introduction HTS Hardware Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cytometer Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cytometer Interface Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Sample Coupler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Sample Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chapter 2: BD FACSDiva Software Overview Workspace Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Plate Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Setup View Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Setup View Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Analysis View Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Analysis View Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chapter 3: Running Samples Starting Up . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Setting Up for Plate-Based Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . ix x xi 13 14 16 19 19 21 23 27 28 32 33 43 48 51 55 56 59

v

Creating Experiments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Creating a Folder and an Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . Setting Up the Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Setting Up the Worksheet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Exporting a Plate as a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Assigning Keywords . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preparing for Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Optimizing Cytometer and HTS Settings . . . . . . . . . . . . . . . . . . . . . . . . Performing Compensation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Acquiring Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pausing the BD High Throughput Sampler . . . . . . . . . . . . . . . . . . . . . . . Stopping the BD High Throughput Sampler . . . . . . . . . . . . . . . . . . . . . . Exporting an Experiment as a Template . . . . . . . . . . . . . . . . . . . . . . . . . Analyzing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Displaying Keywords . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Performing Data Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Maintaining Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Returning to Tube-Based Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Returning to Tube-Based Acquisition on the BD LSR II . . . . . . . . . . . . . Returning to Tube-Based Acquisition on the BD FACSCanto and BD FACSCanto II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Shutting Down . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Quitting the Software . . . . . .