BD FACSAria III Addendumto the BD FACSAria II User’s Guide · PDF file X-mount optical...

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Transcript of BD FACSAria III Addendumto the BD FACSAria II User’s Guide · PDF file X-mount optical...

  • bdbiosciences.com 648240 Rev. A 1/2010

    For Research Use Only. Not for use in diagnostic or therapeutic procedures.

    Becton, Dickinson and Company BD Biosciences San Jose, CA 95131 Tel 877.232.8995 Fax 800.325.9637 [email protected]

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    BD FACSAria III Addendum to the BD FACSAria II

    User’s Guide Including the 561-nm and 445-nm

    Laser Options

  • Copyrights and trademarks

    © 2010, 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.

    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.

    Alexa Fluor® and Texas Red® are registered trademarks and Pacific Blue™ is a trademark of Molecular Probes, Inc.

    Living Colors® is a registered trademark of Clontech.

    BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. © 2010 BD

    FCC information

    WARNING: Changes or modifications to this unit not expressly approved by the party responsible for compliance could void the user’s 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 numérique de la classe A respecte toutes les exigences du Réglement sur le matériel brouilleur du Canada.

    History

    Revision Date Change made

    648240 Rev. A 1/2010 New document

  • Contents

    About this addendum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

    BD FACSAria III flow cell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

    561-nm and 445-nm laser options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

    Laser power switches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

    Laser and optics configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

    Detector arrays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

    Additional system configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

    Special considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

    Accessing the additional octagon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

    Setting up configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

    Typical workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

    Optics part numbers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

  • 1

    About this addendum

    About this topic This topic describes the purpose of this document and the intended audience.

    Purpose The addendum serves multiple purposes.

    This addendum covers the new features for the BD FACSAria™ III cell sorter. This addendum is intended to be used in conjunction with the BD FACSAria II User’s Guide.

    This addendum is also for BD FACSAria and BD FACSAria II customers who have upgraded their systems with the 561-nm or 445-nm laser options. An upgraded BD FACSAria II becomes a BD FACSAria IIIu.

    Features described The BD FACSAria III cell sorter includes the following features, even if the system does not receive a 561-nm or 445-nm laser option.

    The X-mount optical plate

    An optical alignment that includes a fourth beam spot for future upgrading to additional lasers

    A BD FACSAria III flow cell

    An octagon detector array for the 405-nm and 375-nm lasers

    Safety The content of the BD FACSAria II Safety and Limitations Manual applies to the BD FACSAria III cell sorter. Make sure that everyone working with the system is familiar with the safety details in the Safety and Limitations Manual that is shipped with each system.

  • BD FACSAria III Addendum to the BD FACSAria II User’s Guide2

    BD FACSAria III flow cell

    About this topic The topic describes the BD FACSAria III flow cell and the integrated nozzles that are used with it.

    The BD FACSAria III system includes a new flow cell that has been optimized for four laser beam spots. This flow cell is also included with the 375-nm laser option for side population studies.

    Integrated nozzles for the flow cell

    The BD FACSAria III flow cell requires integrated nozzles that have been optimized for it. The system is shipped with these nozzles. These nozzles have also been optimized for BD FACSAria II systems equipped with a 375-nm laser. The following table lists the part numbers for the nozzles, in case you need to order replacement nozzles.

    The nozzles can be identified by the MS marking on the bottom side of the handle. See the following figure.

    The 130-micron and closed-loop nozzles are the same as those used in the BD FACSAria II system. See the user’s guide for those part numbers.

    Nozzle size Part number

    70 micron 647339

    85 micron 647340

    100 micron 647341

    MS indicates integrated nozzle for the BD FACSAria III flow cell

    Bottom view of nozzle

  • 3

    561-nm and 445-nm laser options

    About this topic This topic describes the laser options and the X-mount optical plate.

    Either one or both of these lasers can be added at the factory as an option on a new instrument, or they can be installed by a BD field service engineer into an existing system at your facility.

    561-nm laser option The 561-nm laser can be installed on a system to enable improvements in multicolor experiments and to expand the range of applications for these platforms. Specifically, this option increases PE sensitivity and minimizes or eliminates FITC spectral overlap into the PE channel, giving the instrument the best chance at resolving a dim staining PE population. In addition, this laser improves resolution for mCherry fluorescent proteins.

    445-nm laser option The 445-nm laser can be installed on a system to enable enhanced sensitivity when working with Cyan fluorescent proteins. This laser shares the detector collection optics with the 488-nm laser. If both lasers are installed on a system, you can use either the 445-nm laser or the 488-nm laser, but you cannot use both at the same time. The laser power switches are designed to select one laser or the other.

  • BD FACSAria III Addendum to the BD FACSAria II User’s Guide4

    X-mount optical plate The fiber optic heads for the new lasers and all existing lasers (except the 375 nm) are mounted on an X-mount optical plate. The following figure shows the location for each fiber optic head.

    Fourth beam spot A fourth spatially-separated beam spot is added with the 561-nm laser option. This additional beam spot enables the simultaneous use of four lasers.

    Changes to laser delays If you are an existing BD FACSAria or BD FACSAria II customer, you will notice a difference in default laser delays with your 561-nm or 445-nm laser upgrade compa