BD FACSCanto Options Reference Manual

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http://www.bdbiosciences.com/ Part No. 338628 Rev. A September 2004 For In Vitro Diagnostic Use BD Biosciences 2350 Qume Drive San Jose, CA 95131-1807 USA Tel (877) 232-8995 Fax (408) 954-2347 Brazil Tel (55) 11-5185-9995 Fax (55) 11-5185-9895 Europe Tel (32) 53-720211 Fax (32) 53-720450 Mexico Tel (52) 55-5999-8296 Fax (52) 55-5999-8288 Japan Nippon Becton Dickinson Company, Ltd. Tel 0120-8555-90 Asia Pacific Fax (65) 6-860-1590 Canada Tel (888) 259-0187 Fax (905) 542-9391 Tel (65) 6-861-0633 (905) 542-8028 [email protected] BD FACSCanto Options Reference Manual

Transcript of BD FACSCanto Options Reference Manual

Page 1: BD FACSCanto Options Reference Manual

http://www.bdbioscienPart No. 338628 Rev. ASeptember 2004

For In Vitro Diagnostic

BD Biosciences2350 Qume DriveSan Jose, CA 95131-1807USATel (877) 232-8995Fax (408) 954-2347

BD FACSCantoOptions Reference

Manual

ces.com/

Use

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

Europe Tel (32) 53-720211Fax (32) 53-720450

MexicoTel (52) 55-5999-8296Fax (52) 55-5999-8288

JapanNippon Becton Dickinson Company, Ltd.Tel 0120-8555-90

Asia Pacific

Fax (65) 6-860-1590

CanadaTel (888) 259-0187

Fax (905) 542-9391

Tel (65) 6-861-0633(905) 542-8028

[email protected]

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© 2004, 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 © 2004, 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.

BD FACSCanto™ software © 2004, 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.

BD, the BD logo, BD FACS, BD FACSCanto, BD FACSDiva, BD Falcon, and BD Trucount are trademarks of Becton, Dickinson and Company.

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, can 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 the matériel brouilleur du Canada.

Notice

BD Biosciences delivers software and workstations that are intended for running the instruments 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/user’s 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

History

Revision Date Change Made

336929 Rev. A 1/04 Initial release

337949 Rev A 4/04 Updated for CE IVD release

338628 Rev A 9/04 Updated for BD FACSDiva 4.1 release

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Contents

BD FACS Loader Option 5

Loader Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

Loader Software Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Carousel Preferences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Lifter Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

Carousel Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

Using the Loader . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Starting Up the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Setting Up a Worklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

Importing a Worklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Assigning Carousels and Verifying Run Settings . . . . . . . . . . . . . . . . . . . 19

Preparing the Loader . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

Running Samples with the Loader . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Changing Operational Modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Maintaining the Loader . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Cleaning External Surfaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

Technical Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Supplies and Replacement Parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

Index 41

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BD FACS Loader Option

The BD FACS™ Loader introduces prepared samples automatically to the BD FACSCanto™ flow cytometer. The Loader consists of a drawer, a cover, two optical sensors, an electronics module, a tube lifter, and a 40-tube carousel. You can operate it with BD FACSDiva™ software or BD FACSCanto™ clinical software.

This guide contains the following information:

• Loader Components on page 6

• Loader Software Controls on page 8

• Using the Loader on page 14

• Maintaining the Loader on page 33

• Troubleshooting on page 35

• Technical Specifications on page 38

• Supplies and Replacement Parts on page 39

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Loader Components

The Loader is mounted directly on the flow cytometer. The device consists of a drive system, tube lifter mechanism, spindle, and optical sensors, all of which are attached to a sliding drawer. A cover fits over this drawer to protect you from moving parts during operation. Table 1 provides a brief description of Loader components.

Table 1 Loader components

optical sensors (2) • reads the carousel ID

• scans the carousel to verify tube locations and match the associated worklist, and verifies a tube is in place before activating the tube lifter

drawer holds the carousel; slides out for easy access

center spindle the axis around which the carousel spins; insert the carousel onto the Loader here

alignment guide pin correctly lines up the carousel and keeps it in place

tube lifter a stainless steel rod that lifts the sample tube from the carousel to the sample injection tube (SIT)

drive system the motor that rotates the carousel (not visible from exterior)

center spindlealignment guide pin

optical sensors

tube lifterdrawer

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Carousel

The carousel accommodates up to forty 12 x 75-mm tubes. Each carousel has a unique ID printed on its top surface and on an optically read label on its interior surface.

Figure 1 Sample-Prep Ready carousel

Note that the Loader is compatible with both the green-tinted carousels labeled “Sample-Prep Ready” and the traditional gray Loader carousels (Figure 2).

Figure 2 Compatible carousels

Not all manufactured 12 x 75-mm test tubes have been checked for proper functionality on the Loader. BD Biosciences has validated only disposable, 12 x 75-mm BD Falcon™ polystyrene test tubes and BD Trucount™ tubes.

alignment hole

spindle hole

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Loader Software Controls

The following Loader controls are available in BD FACSDiva software. For a description of Loader software controls in BD FACSCanto clinical software, refer to the BD FACSCanto Clinical Software User’s Guide.

• Carousel preferences can be accessed from the Edit > User Preferences menu. For a description, see the following section, Carousel Preferences.

• Carousel setup and run settings are located in the Carousel Setup dialog box, accessed by choosing Experiment > Carousel Setup. See Assigning Carousels and Verifying Run Settings on page 19.

• Loader acquisition controls can be found in the Carousel Controls frame. These controls are described in Running Samples with the Loader on page 25.

• Lifter settings are shown on the Lifter tab in the Instrument frame. See Lifter Settings on page 12.

A Carousel Report is generated when a run is complete or stopped in process. A report description can be found in Carousel Report on page 12.

Carousel Preferences

To access Preferences for the Loader, choose Edit > User Preferences, and click the Carousel tab. There are two tabs under the Carousel tab: General and Save Options. The General tab contains preferences for printing and reviewing worksheets. The Save Options tab contains preferences for saving statistics and the Carousel Report.

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General Preferences

Use General preferences to specify automatic printing preferences and how long to review a worksheet before proceeding with the next Tube.

During Loader acquisition, data is shown on a global worksheet. Select Automatic Printing Options for the following.

• To print a copy of the global worksheet after each Tube is processed, select the Worksheet checkbox, and then click Print after each tube.

• To print a copy of the global worksheet after each Specimen is processed, select the Worksheet checkbox, and then click Print after each specimen. Analysis objects for all Tubes in a Specimen are accumulated onto a worksheet, which prints when the last Tube in the Specimen is processed.

• To print a carousel report after a run, select the Carousel report checkbox. For more information about this report, see Carousel Report on page 12.

Printouts are sent to your default Windows® printer, unless you print manually and use the Print Setup dialog box to choose a different printer.

! Tip If your Specimen and Tube names combined have more than 80 characters, set the printer to landscape mode.

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Select an option to specify how long you want to review the global worksheet before the cytometer proceeds with acquisition of the next Tube.

• To pause after the first Tube in each panel (or Specimen), click Pause after first tube. Data is displayed on the global worksheet until you click Resume.

• To pause after each Tube, click Pause after each tube. Select Auto to have acquisition of the next Tube proceed automatically after a specified period of time (1–10 seconds), or Manual to pause until you click Resume.

Save Options Preferences

Use Save Options preferences to set up automatic saving options for statistics and the Carousel Report. You can specify to save statistics results, a Carousel Report, or both, and use the Folder Location field to specify where each file will be saved.

! Tip Select the Date folder checkbox to automatically create a dated folder in the specified directory for the corresponding exported reports.

A Statistics Results file is formatted with a horizontal orientation. For each Specimen, statistics view(s) header information is written to the file for the first

If you select Pause after each tube and set the amount of time to zero, you will not be able to review the worksheet between Tubes.

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Tube. (When there is more than one statistics view for a Tube, an empty column is used to separate header information for each view.) Header information comprises the following:

• Experiment Name

• Specimen Name

• Tube Name

• Collection Date

• Selected keywords

• Statistics label

As each Tube in the Specimen is recorded, a new row of results is added to the file. For each population in a Tube’s statistics view(s), the application adds parameter statistics in the order in which they appear in the statistics view. (Output is padded with commas so that the exported statistics views line up appropriately in the csv output.) A new header row is added only if statistics in the statistics view(s) differ from one Tube to the next within the Specimen.

If you elect to skip a Tube before recording begins, no statistics are written to the file. If the Tube is skipped after recording begins but before recording is complete, statistics for the recorded data will be written. If the Specimen or Tube is rerun, the application adds a new row of results as each Tube is recorded.

If you add or remove statistics from a statistics view for a Tube while the carousel is running, the application writes a new row of headers to the export file. The remaining Tubes (unless there is another statistics change) will display the new set of statistics.

When there are duplicate results for a Tube or Specimen, check the date/time stamp to see which are the most recent results.

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Lifter Settings

Lifter settings are used to adjust the tube lifter to ensure proper sealing with the SIT. You might be asked to communicate these settings to BD Biosciences personnel if you call for troubleshooting assistance. You can change or view Lifter settings only if you are logged into BD FACSDiva software as an Administrator.

Carousel Report

When a run is complete or stopped, a report is generated with information about the Tubes in the carousels. The Carousel Report is displayed in a separate window with a menu bar above the report header. The report is automatically saved or printed or both if the corresponding preference is selected. The following information is included:

• Experiment name, and the Specimen and Tube name for each tube on the carousel

• current date and time

• User ID (login name)

Do not change Lifter settings unless directed to do so by BD Biosciences.

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• instrument name and serial number

• carousel number and position of each tube

• FCS file names and location

• statistics file name and location (if generated)

• Tube collection date/time

• status of each Tube

To print the report, choose File > Print Report within the Carousel Report window. You can preview the report or set up the page for printing by choosing the corresponding selections from the File menu.

! Tip If your Specimen and Tube names combined have more than 80 characters, set the printer to use landscape mode.

If you did not select the automatic export preference, you can export a report by choosing File > Export within the Carousel Report window. A dialog box appears where you can specify the file storage location. Exported reports are

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saved as an XML file that can be opened and printed using Microsoft Internet Explorer.

Using the Loader

The Loader operates through software commands only. Commands can be accessed through BD FACSDiva software or BD FACSCanto clinical software. For instructions on running samples with BD FACSCanto clinical software, refer to the BD FACSCanto Clinical Software User’s Guide.

This section explains how to acquire samples with the Loader using BD FACSDiva software commands. You should have a working knowledge of BD FACSDiva software before you run the following procedures:

• Starting Up the Instrument on this page

• Setting Up a Worklist on page 15

• Assigning Carousels and Verifying Run Settings on page 19

• Preparing the Loader on page 22

• Running Samples with the Loader on page 25

Starting Up the Instrument

Use the following steps as a guideline to get ready for acquisition. For more details, refer to the BD FACSCanto Flow Cytometer Reference Manual.

1 Start up the cytometer, the workstation, and the software as described in your instrument manual.

2 Perform instrument quality control and sample optimization for the samples you will be running.

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3 (Optional) Open a new Experiment, and copy your optimized instrument settings from step 2 to the new Experiment.

Alternatively, you can add Specimens to the Experiment that was used during sample optimization.

Setting Up a Worklist

To run samples on the Loader, you must first add Specimens and Tubes to your Experiment. Alternatively, you can import Specimen and Tube information from a worklist created using BD FACS™ Sample Prep Assistant (SPA) software. See Importing a Worklist on page 17 for instructions.

The Carousel view of your Experiment becomes your worklist.

You must use a global worksheet for data display when you are acquiring samples with the Loader.

To prevent data loss, do not create Experiments that contain more than 50 Tubes.

worklist

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1 Add the required number of Specimens and Tubes to your Experiment.

! Tip To fit the combined Specimen and Tube names on the Carousel Setup Report, keep them less than 80 characters long. Also, make the first 40 characters of each name unique.

To import a predefined panel, do the following:

• Select any Browser item within the open Experiment, and choose Experiment > New Specimen.

• Click a tab in the Panel Templates dialog box, and select a panel by name.

• Specify the number of copies (each copy creates a new Specimen in the Browser), and click OK.

For more details about panel templates, refer to the BD FACSDiva Software Reference Manual.

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2 If you imported panels, inspect the instrument settings for each panel type to make sure the settings are appropriate.

Importing a Worklist

Use the following steps to import Specimen and Tube information from a Worklist created in BD FACS SPA software.

1 Choose File > Import > Worklist.

Note that sample information can be imported only into a new Experiment. If you try to import a worklist into an existing Experiment, the software automatically closes the current Experiment and creates a new one during the import.

2 Locate and select a file to import in the dialog that appears; click Import (Figure 3 on page 17).

Figure 3 Selecting a file to import

Note that only valid SPA worklists can be imported. Panels in the imported worklist must match the panels available in BD FACSDiva software, and each imported panel must have the same number of Tubes as those in the

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predefined panels. If any of these criteria are not met, an error message appears.

After a successful import, a new Experiment is created with the SPA worklist name as the Experiment name. Note that periods or commas in the SPA worklist name are changed to an underscore (_) in the Browser. If <SampleIDs> are used in the SPA worklist, Specimens are named according to the <SampleID> information for each panel.

3 Review the imported information in the Browser; edit missing or incorrect entries, if needed.

Inspect the instrument settings for each panel type to make sure the settings are appropriate.

4 Continue to the next task, Assigning Carousels and Verifying Run Settings on page 19.

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Assigning Carousels and Verifying Run Settings

Once you set up an Experiment with required Specimens and Tubes, you must assign Specimens to a carousel using the Carousel Setup dialog box.

1 Choose Experiment > Carousel Setup.

The Carousel Setup dialog box appears listing all Specimens in the Experiment that do not contain recorded data. Carousel locations are automatically assigned according to the number of Tubes per Specimen.

2 (Optional) Insert or delete carousel breaks.

A carousel break is represented by a thick horizontal line, and indicates the start of a new carousel. Breaks are inserted automatically when a carousel cannot contain the next Specimen’s Tubes. (You cannot split a Specimen between two carousels.) If you want to group Tubes differently from how they are assigned to the carousels, you can insert and delete breaks.

You can insert or delete breaks only on an unassigned carousel.

• To insert a break, select an unassigned Specimen and click Insert Break.

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• To delete a break, select an unassigned Specimen under a break and click Delete Break. Note that you can delete a break only if the preceding carousel has room for all the Tubes assigned to the current carousel.

3 Assign a carousel to each Specimen.

BD carousels are pre-labeled with carousel IDs, from 1 to 16, both on top and on an optically read label inside. Click in the Carousel field next to the Specimen name and choose a carousel ID from the drop-down menu that appears. All Specimens assigned to the same carousel are automatically given the same carousel ID.

If you imported sample information from a SPA worklist, carousel and Tube positions are read from the imported worklist and cannot be changed. If <CarouselID> was not defined in the imported worklist, the carousel column is set to None and you must assign carousels before you can run.

4 Click the Print button on the Carousel Assignment tab to print a record of Tube and carousel assignments.

For accurate results, match the tubes to those listed on the Carousel Assignment tab. Print out a copy of tube assignments and use the printout as a guide when filling each carousel.

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5 Click the Run Settings tab in the Carousel Setup dialog box, and verify the settings.

Use the Run Settings tab to specify the amount of time to acquire samples before recording begins (Acquisition time), and to enter mix settings.

• Choose an Acquisition time, from 3–15 seconds. The acquisition time allows acquisition to stabilize before recording begins.

• Select the Start of carousel mix checkbox if you want to perform a 10-second mix before the start of each carousel.

• Select the Interim mix checkbox if you want to perform a 3-second mix between a specified number of tubes. After you select the checkbox, the drop-down menu is activated where you can choose how often you want mixing to occur, from 1–20 tubes.

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Preparing the Loader

Once you have prepared a worklist, follow these steps to set up the cytometer for acquisition with the Loader.

1 Verify that the cytometer is set up for automatic loading.

The adapter lever should be in the lowest position, the tube guide should be forward, and the aspirator arm bar should be horizontal (Figure 4).

Figure 4 Automatic loading setup

For detailed setup instructions, see Changing to Automatic Loading on page 32.

adapter lever

tube guide

aspirator arm bar

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2 Remove the carousel from the instrument, if needed.

Slide the Loader cover forward and pull out the Loader drawer. Place your thumb on the center spindle, and press down with your thumb while gently pulling up on the carousel handle with two fingers, as shown.

3 Vortex the sample tubes and place them uncapped in the carousel(s) according to the worklist.

4 Install the first carousel to be acquired on the Loader.

Position the spindle hole in the handle over the center spindle of the Loader drawer. Rotate the carousel until the alignment guide pin fits into the small alignment hole at the top of the carousel. Press down firmly to seat the carousel. See Figure 5 on page 24.

For accurate results, match the tubes to those listed on the Carousel Assignment tab. Print out a copy of tube assignments and use the printout as a guide when filling each carousel.

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Figure 5 Seating the carousel in the Loader

5 Close the Loader drawer completely, and install the Loader cover.

The Loader scans and positions the carousel at tube 1.

To run the Loader, the cover must be in place on the drawer. Tubes will not be loaded if the cover is off, and currently running tubes will be unloaded if the cover is removed during a run.

handle

spindle

alignmentpin

Loader cover

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Running Samples with the Loader

1 In the Acquisition Controls frame, verify that you are using an appropriate Flow Rate.

2 In the Carousel Controls frame, enter a stopping Time (seconds).

If the Time is 0, the software uses a default stopping gate based on number of events. When you enter a Time, the software stops either when it reaches the total number of events, or when it reaches the Time limit, whichever comes first.

Setting a Time limit ensures that the carousel doesn’t stall on one tube if that tube should contain a dilute sample or rare events.

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3 In the Carousel Controls frame, click Run.

To display the frame, click the Carousel Controls button in the Workspace toolbar ( ).

The Loader performs the following initialization steps:

• a start-of-carousel mix, if one was specified in Run Settings

• carousel ID check

• tube location check against the Carousel Assignment tab

4 View data for the first Tube on the global worksheet.

The Carousel Controls frame displays a Pause button and a countdown message showing the time remaining until recording.

If needed, click the Pause button, make necessary instrument settings adjustments, and then click Resume.

Data is recorded for the first Tube. Recording continues until the stopping rule is met. The Carousel Controls frame shows the ID of the currently running carousel along with the Tube’s position and name.

tube namecarousel ID

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During recording, you can skip the Tube or Specimen, rerun the Tube or Specimen, or stop the run. For more information, see Skipping or Re-Running Samples on page 28 or Stopping a Run on page 29.

5 Make adjustments to the analysis, if needed.

Data for the current Tube remains on display in the global worksheet view for the amount of time specified in General Carousel preferences. During this time, you can adjust the analysis, manually print the worksheet, skip the Tube or Specimen, rerun the Tube or Specimen, stop the run, or pause. A Pause button appears if you chose a manual pause after each tube (Figure 6). In the example shown in Figure 6, no Pause button will appear in the Carousel Controls.

Figure 6 Selections that affect pausing

If you selected the Print after each tube preference, the worksheet prints automatically.

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6 Click Resume (if necessary) to proceed with acquisition of the next Tube, or let the timer count down to zero.

Note that any changes to the global worksheet remain in effect for the rest of the Tubes that use the same worksheet.

Tube processing continues until the carousel is complete. Depending on how you set Carousel preferences, review the global worksheet after the first Tube in each panel (or Specimen), or after each Tube. Repeat step 6 for the remaining Tubes on the carousel.

If specified in run settings, mixing occurs on an interim basis.

7 Load the next carousel, if applicable.

If multiple carousels have been assigned, the software prompts you to load the next carousel.

8 Click Continue.

9 When all carousels have been processed, review the Carousel Report.

The report prints automatically, if specified in General Carousel preferences. If you selected automatic saving in the Save Options preferences, the report is saved to the designated directory.

For more information, see Carousel Report on page 12.

Skipping or Re-Running Samples

You can skip or re-run Tubes or Specimens in the following situations:

• during acquisition, when the recording countdown is paused

• during recording

• when processing of the current Tube is finished and the global worksheet view is paused

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To skip or re-run a sample, click the corresponding button in the Carousel Controls frame. A drop-down menu appears where you can choose to Skip Tube/Skip Specimen or Rerun Tube/Rerun Specimen.

When you skip a Tube, the carousel starts processing the next Tube in the worklist; when you skip a Specimen, the carousel starts the first Tube under the next Specimen. If there are no more Tubes to process, the Carousel Report appears.

When you rerun a sample, existing data is not overwritten.

• If the Export FCS after recording preference is enabled, a new FCS file is saved as: <Specimen Name>_<Tube Name>_<DDMMYYYYHHMMSS>.fcs, where DDMMYYYY represents the date and HHMMSS represents the time the new file was saved.

• If the Automatic Saving preference is enabled for statistics results, a new row is written to the statistics results file. See Save Options Preferences on page 10 for more information.

Stopping a Run

To stop processing a carousel, click Stop in the Carousel Controls frame. The carousel stops running and the Carousel Report appears.

If a Tube is being processed when you click Stop, the tube is unloaded from the cytometer.

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Changing Operational Modes

Performing a few simple steps allows you to change from automatic loading to manual loading. See the following sections for instructions.

• Changing to Manual Loading

• Changing to Automatic Loading on page 32

Changing to Manual Loading

To change to manual mode, follow these steps.

Materials

personal protective equipment

Procedure

1 Remove the Loader cover.

2 Pull out the drawer.

3 Remove any carousel in the Loader.

4 Move the tube guide arm all the way back and away from you.

Handle all biological specimens and materials as if capable of transmitting infection. Dispose of waste using proper precautions and in accordance with local regulations. Wear suitable protective clothing, eyewear, and gloves.

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When in manual mode, you do not need the tube guide arm.

5 Lift the adapter lever and rotate it into the highest position, as shown in the following figure.

6 Rotate the aspirator arm bar vertically.

automatic mode manual mode

adapter lever, automatic mode

adapter lever, manual mode

automatic mode manual mode

Using the Loader 31

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You are ready to load tubes manually.

Changing to Automatic Loading

To change from manual loading to automatic loading, follow these steps.

Materials

personal protective equipment

Procedure

1 Lift the adapter lever and rotate it into the automatic mode position, as shown in the following figure.

Handle all biological specimens and materials as if capable of transmitting infection. Dispose of waste using proper precautions and in accordance with local regulations. Wear suitable protective clothing, eyewear, and gloves.

adapter lever, automatic mode

adapter lever, manual mode

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2 Rotate the aspirator arm bar to the horizontal position until it reaches the stop pin.

3 Move the tube guide arm all the way forward.

The tube guide arm is needed only when using the Loader.

4 Pull out the drawer.

You are ready to load tubes automatically.

Maintaining the Loader

This section explains the cleaning procedure you should follow to keep your Loader in good condition.

manual mode automatic mode

manual mode automatic mode

Maintaining the Loader 33

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Cleaning External Surfaces

To keep the Loader free from salt buildup, wipe down all external surfaces that have been exposed to sheath fluid.

Materials

• BD FACS™ cleaning solution

• DI water

• clean, lint-free cloths or disposable wipes

Procedure

1 Switch off the cytometer power and unplug the AC power cord from the wall socket.

2 Wipe all accessible surfaces with BD FACS cleaning solution.

Make sure that you clean the optical sensors, as well. Salts can build up on the sensors, causing tubes in the carousel to be missed.

3 Wet a fresh cloth with DI water and wipe all exposed surfaces to prevent corrosion.

4 Wipe all exposed surfaces with a clean, dry cloth.

All instrument surfaces that come in contact with biological specimens can transmit potentially fatal disease. Use universal precautions when cleaning instrument surfaces. Wear suitable protective clothing and gloves.

To prevent damage, do not use isopropyl alcohol or ethanol on any Loader surfaces.

To avoid potential shock, always switch off the power and unplug the AC power cord before you begin cleaning.

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Troubleshooting

The tips in this section are designed to help you troubleshoot Loader problems. If additional assistance is required, contact your local BD Biosciences technical support representative. Refer to our website, http://www.bdbiosciences.com, for up-to-date contact information.

For software problems, refer to the BD FACSDiva Software Reference Manual or to the BD FACSCanto Clinical Software User’s Guide.

Observation Possible Causes Recommended Solutions

Carousel not rotating correctly

Carousel not engaged with alignment pin on drawer

Rotate the carousel on the spindle until the alignment guide pin engages with the alignment hole in the carousel, and press down. See step 4 on page 23. If the problem persists, contact BD Biosciences for assistance.

Aspirator arm bar vertical Toggle the bar until it is horizontal. See Changing to Automatic Loading on page 32.

Troubleshooting 35

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Test tube not loading Wrong tubes used Use only uncapped 12 x 75- mm BD Trucount tubes or polystyrene BD Falcon tubes.

Bal seal not properly installed

Reinstall according to the instructions given in Replacing the Bal Seal in the BD FACSCanto Flow Cytometer Reference Manual.

Bal seal worn Replace the Bal seal. Refer to Replacing the Bal Seal in the BD FACSCanto Flow Cytometer Reference Manual.

Loader not prepared for automatic loading

Modify the Loader according to Changing to Automatic Loading on page 32.

Test tube not completely raised

Make sure the Loader drawer is completely shut.

Make sure the test tube is free of unnecessary labels or tape.

Tube lifter hitting carousel during ascent

Make sure the carousel is properly engaged with the alignment guide pin. If the problem persists, contact BD Biosciences for assistance.

Tube lifter failure Contact BD Biosciences.

Test tube missed, or missing tube messages in software

Salt crystal buildup on optical sensors

Clean the optical sensors, as described in Cleaning External Surfaces on page 34.

Test tube runs dry, Loader not advancing to next sample

Dilute sample or rare events

Set a stopping Time. See step 2 on page 25.

Observation Possible Causes Recommended Solutions

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Test tube stuck on SIT, software not responding

Software problem Manually remove the tube from the SIT.

1 Hold the tube while you move the aspirator arm to the side.

2 Gently pull the tube off the SIT.

3 Release the aspirator arm.

4 Restart the software.

Test tube stuck on SIT Not configured for automated loading

Manually remove the tube from the SIT.

1 Hold the tube while you move the aspirator arm to the side.

2 Gently pull the tube off the SIT.

3 Release the aspirator arm.

4 Configure the instrument for automatic loading. See Changing to Automatic Loading on page 32.

Observation Possible Causes Recommended Solutions

Troubleshooting 37

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Technical Specifications

Carousel compatibility Loader carousels, numbers 1–16

Tube compatibility

• Carousel Accommodates up to 40 uncapped 12 x 75-mm tubes

• BD Falcon polystyrene test tubes

• BD Trucount tubes

• BD FACS 7-color setup bead tubes

• Thickness of accumulated labels ≤5 mils (127 µm) thick

Tube sample volume <1 mL

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Supplies and Replacement Parts

This section provides a list of supplies and options that are available for the BD FACSCanto instrument.

• To order spare parts and consumables from BD Biosciences from within the US, call: (877) 232-8995 or go to www.bdbiosciences.com.

In other countries, contact your local BD Biosciences representative.

• To order instrument options, contact your sales representative.

This information is correct at the time of publication; for up-to-date information refer to our website (http://www.bdbiosciences.com).

Item Part No.

Carousel kit, carousels 1–4 332727

Carousel kit, carousels 5–8 332728

Carousel kit, carousels 9–12 332729

Carousel kit, carousels 13–16 332730

BD Falcon 12 x 75-mm tubes (bag of 125) 343675

Loader cover replacement 335981

Supplies and Replacement Parts 39

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Index

A

acquiring with Loader 25acquisition, stopping

manual stop, carousel 29administrator screens 12alignment guide pin 6automatic loading setup 22, 32

C

Carousel Controls frame 25, 26carousel rack

compatible racks 7described 7installing 23mixing preferences 21problems with 35removing 23software setup 19tubes, compatible 7, 38

carousel report 10, 12cleaning 34components 6cover 6, 24, 39cytometer

automatic (Loader) mode 22, 32manual mode 30startup 14

D

decontamination 34drawer 6dry tube 36

F

framesCarousel Controls 25, 26Instrument 12

G

guide pin 6

I

installing carousel rack 23

L

label thickness 38lifter, tube 6, 12Loader

maintenance 34

M

maintenanceLoader 34

manual loading mode 30

41

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mixing preferences 21modifying cytometer 22, 30, 32

N

names, number of characters 13

O

optical sensors 6ordering spare parts 39

P

parts, ordering 39pause preferences 27pin, alignment 6preferences

global worksheet 10length of review time 10mixing 21overview 8pausing 27printing 8, 9saving 10tube lifter settings 12worksheet 8

printing preferences 8, 9

R

rack, carousel 38removing carousel rack 23re-running samples 28running samples 25

S

Sample Prep Assistant (SPA)panels, valid 17worklists 15, 17

save preferences 10skipping samples 28specifications 38Specimen names 13spindle 6statistics report 10Statistics Results file 10stopping acquisition

manual stop, carousel 29supplies, ordering 39switching modes

automatic (Loader) to manual 30manual to automatic (Loader) 32

T

technical specifications 38troubleshooting 35Tube names 13tubes

compatible 7lifter 6, 12sample volume 38

U

user preferences for Loader 8

W

windowsCarousel Controls 25, 26Instrument 12

worklistimporting 17setting up 15

worksheet preferences 8, 10

42 BD FACSCanto Options Reference Manual