BD Accuri C6 User manual

BD Accuri C6 Flow Cytometer Instrument Manual
7820018 Rev-2 i
TABLE OF CONTENTS
1INTRODUCTION .................................................................................................................1
1.1 About This Manual.....................................................................................................1
1.2 Delivery and Inspection .............................................................................................1
1.3BD Accuri C6 Flow Cytometer Specifications.............................................................2
2SAFETY PRECAUTIONS ....................................................................................................3
2.1 Safety and Limitations Manual...................................................................................3
2.2 Operational Considerations .......................................................................................3
2.2.1 Ambient Temperature and Humidity...............................................................3
2.2.2 Area of Use....................................................................................................3
2.2.3 Approved Reagents .......................................................................................3
2.2.4 Material Safety Data Sheets (MSDS).............................................................4
3SETTING UP THE BD ACCURI C6 FLOW CYTOMETER ...................................................5
3.1 BD Accuri C6 Flow Cytometer Layout........................................................................5
3.2 Configuring the Computer Settings............................................................................5
3.2.1 Setting up the Computer ................................................................................5
3.2.2 Changing the Workstation Computer Settings for Windows®7.......................6
3.2.3 Viewing .c6 and .c6t File Extensions in Windows 7........................................6
3.3 Setting Up the BD Accuri C6 Flow Cytometer............................................................7
3.3.1 Setting Up the Reagent Bottles and Fluidics ..................................................8
3.3.2 Connecting the Cables...................................................................................9
3.4 Installing Software ...................................................................................................10
3.4.1 Installing BD Accuri C6 Software..................................................................10
3.5 Starting Up the BD Accuri C6 Flow Cytometer.........................................................10
3.6 Turning Off the BD Accuri C6 Flow Cytometer.........................................................11
4MAINTENANCE.................................................................................................................13
4.1 Automated Preventive Maintenance Notification......................................................13
4.2 Cleaning the SIP......................................................................................................14
4.3 Cleaning the Flow Cell.............................................................................................14
4.3.1 Running the Unclog Cycle............................................................................14
4.3.2 Running an Extended Clean of the Flow Cell...............................................15
4.4 Decontamination Fluid Cycle ...................................................................................15
4.5 Cleaning Fluid Cycle................................................................................................15
4.6 Purging Air from the BD Accuri C6 Flow Cytometer.................................................16
4.7 Emptying the Waste Container ................................................................................16
4.8 Filling the Fluid Bottles.............................................................................................17
4.9 Replacing the Fluidic Bottle Filters...........................................................................17
4.10 Inspecting the Fluidics Lines....................................................................................18
4.11 Replacing the In-line Sheath Filter...........................................................................18
4.12 Replacing the Peristaltic Pump Tubing ....................................................................19
5TROUBLESHOOTING AND TECHNICAL SUPPORT........................................................21
5.1 Troubleshooting.......................................................................................................21
5.2 Cleaning and Decontamination—Standard Procedure.............................................24
5.3 Limited Warranty .....................................................................................................25
5.4 Extended Warranties...............................................................................................25
5.5 Technical Support....................................................................................................25
5.6 Ordering Information................................................................................................27

BD Accuri C6 Flow Cytometer Instrument Manual
ii 7820018-01 Rev-2
FIGURES AND TABLES
Figure 1-1. The BD Accuri C6 Flow Cytometer System............................................................. 1
Table 1-3. BD Accuri C6 Flow Cytometer Specifications............................................................ 2
Table 2-1. Approved Reagents.................................................................................................. 3
Figure 3-1. BD Accuri C6 Flow Cytometer Optics and Fluidics (Top View, Lid Open)................ 5
Figure 3-2. In-line Sheath Filter................................................................................................. 7
Figure 3-3. Interference Filter.................................................................................................... 7
Figure 3-4. Fluidics harness...................................................................................................... 8
Figure 3-5. Proper Attachment of Fluidics Lines........................................................................ 9
Figure 3-6. Cable Connections.................................................................................................. 9
Figure 3-7 SIP and Sample Stage............................................................................................10
Figure 3-8. Improper Shut down Message................................................................................11
Table 4-1. BD Accuri C6 Flow Cytometer Replacement Parts/Reagents List............................13
Figure 4-1. Automated Preventive Maintenance Notification Window.......................................14
Figure 4-2. Waste Line Quick Disconnect.................................................................................17
Figure 4-3. In-line Sheath Filter................................................................................................18
Figure 4-4. Peristaltic Pumps and Luer Locks ..........................................................................19
Figure 4-5. Removing Peristaltic Pump Retainer Clip...............................................................19
Figure 4-6. Sliding Luer Lock Fittings Off of Pump Head..........................................................20
Figure 5-1. Browse For Folder Dialog Box................................................................................22
Figure 5-2. Update Driver Software –Cytometer, Second Dialog Box......................................22
Figure 5-3. Windows Security Window.....................................................................................23
Figure 5-4. Update Driver Software –Cytometer, Successfully updated your driver software...23
Figure 5-5.The C6 Software Status Message Box –C6 is connected and ready.....................23

BD Accuri C6 Flow Cytometer Instrument Manual
7820018-01 Rev-2 1
The BD Accuri™C6 flow cytometer offers an entirely new vision for the role of flow
cytometry in life science research. Designed from the ground up to be compact and
easy to use, the BD Accuri C6 flow cytometer fits in any lab and is readily accessible
to all researchers—experts and novices alike.
Figure 1-1. The BD Accuri C6 Flow Cytometer System
The BD Accuri C6 flow cytometer system includes BD Accuri C6 Software for
controlling the instrument, generating statistics, and analyzing results.
This manual provides information on cytometer requirements and specifications,
setup, and maintenance. It also includes common troubleshooting help and service
information.
For information on using BD Accuri C6 Software, see the BD Accuri C6 Software
User Guide.
For information on installing and using the BD CSampler™, see the BD Accuri
CSampler Software User Guide.
Carefully inspect all cartons upon receipt of the cytometer. If there are any signs of
mishandling or damage, file a claim with the carrier immediately. If the shipment is
separately insured, file a claim with the insurer.
ITEMS NOT PROVIDED
Surge suppressor or uninterruptible power supply (UPS). A model rated at a
minimum of 1000 VA is recommended for use with the cytometer, computer, and
monitor.
Microsoft®Office™or equivalent
Anti-virus software. This system is intended for use as a stand-alone station, not
as part of a network.

BD Accuri C6 Flow Cytometer Instrument Manual
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Table 1-3. BD Accuri C6 Flow Cytometer Specifications
Laser Excitation
488 nm (rated at 20,000 hr life)
640 nm (rated at 20,000 hr life)
Laser Profile
10 x 75 µm
Laser Power
14.7 mW 640 nm Diode Red Laser
20 mW 488 nm Solid State Blue Laser
Scatter Detection
Forward (0 degrees, +/-13)
Side (90 degrees, +/-13)
Emission Detection
4 colors, standard optical filters
FL1 533/30 nm (e.g. FITC/GFP)
FL2 585/40 nm (e.g. PE/PI)
FL3 > 670 nm (e.g. PerCP, PerCP-Cy™5.5, PE-Cy7)
FL4 675/25 nm (e.g. APC)
Optical Alignment
Fixed alignment
Flow Cell
200 µm ID quartz capillary
Minimum Detectable Particle
Size
0.5 µm
Minimum Sample Volume
300 µL (in 12 x 75 mm tube)
Pre-Set Flow Rates and Core
Sizes
Slow: 14 µL/min, 10 µm core
Medium: 35 µL/min, 16 µm core
Fast: 66 µL/min, 22 µm core
Custom Sample Flow Rates
10 –100 µL/min
Custom Core Diameter
5 –40 µm
Recommended Sheath Fluid
0.22 µm filtered, deionized (DI) water
Maximum Events Per Sample
1 million events per well
Florescence Sensitivity,
MESF*
FITC < 150; PE < 100
Scatter Resolution
Resolves human peripheral blood lymphocytes,
monocytes, and granulocytes
Fluorescence Linearity
2 ± 0.05% for chick erythrocyte nuclei (CEN)
Fluorescence Precision
≤3% CV for CEN
Data Acquisition Rate
10,000 events/second, maximum
Power Requirements
100 - 240 VAC, 50/60 Hz
Typical Power Consumption
154 VA
Heat Output
240 Btu/hour maximum output
Operating Ranges
15 - 30° C; < 80% relative humidity
Instrument Size
11”H x 14.75”W x 16.5”D (27.9 x 37.5 x 41.9 cm)
Foot Print with Fluid Bottles
11”H x 21.5”W x 16.5”D (27.9 x 54.6 x 41.9 cm)
Weight
30 lbs (13.6 kg)
Fluid bottle capacity
2 L sheath fluid, 2 L waste,
250 mL cleaner, 250 mL decontamination
Signal Processing
24-bit datapath
Computer Interface
USB 2.0
Warranty
1 year
*MESF values determined using Thermo Scientific Cyto-Cal™ Multifluor Plus Violet Beads.

BD Accuri C6 Flow Cytometer Instrument Manual
7820018 Rev-2 3
Refer to the BD Accuri C6 Safety and Limitations Manual (PN 7820091) for complete
safety information.
When setting up the cytometer, note the details in the following sections to maximize
the instrument’s performance.
Operating temperature of the cytometer is between 15°C and 30°C (59°F and 86°F).
Operating humidity is < 80% relative humidity.
The cytometer is designed to fit on a laboratory bench top. Keep a perimeter of at
least six (6) inches (15 cm) around the instrument and computer for proper
ventilation and to protect the instrument and computer from liquid spills.
Take special care while handling fluids around the cytometer. Ensure that
uncontained fluids do not enter the instrument or computer. Clean up spills
immediately. Never place anything on top of the cytometer, including sample tubes
and racks.
Turn off the instrument and unplug the power cord before manually cleaning the
cytometer.
Only approved fluids should be used in the operation or cleaning of this instrument.
See the following table for a list of approved reagents.
Table 2-1. Approved Reagents
Fluid
Description
Sheath fluid
0.22 µm filtered, deionized (DI) water with or without
bacteriostatic concentrate solution (PN 653156)
Cleaning solution
Cleaning concentrate solution (PN 653157) diluted
according to package instructions
Decontamination solution
Decontamination concentrate solution (PN 653154)
diluted according to package instructions
Extended flow cell clean
Extended flow cell cleaning solution (PN 653159)
Do not use household bleach as a decontamination solution. It contains
fluorescent whitening agents that may interfere with fluorescence detection.

BD Accuri C6 Flow Cytometer Instrument Manual
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To obtain an MSDS, visit www.AccuriCytometers.com or contact:
In the USA:
Email [e-mail]
Call Accuri Technical Support at 1.734.994.8000
Outside the USA
Visit www.AccuriCytometers.com for country-specific contact information.

BD Accuri C6 Flow Cytometer Instrument Manual
7820018 Rev-2 5
Perform all of the procedures in this chapter before running any experimental
samples.
The cytometer is composed of fluidics, optics, and electronics subsystems that work
together to analyze cell populations. The following figures show the major
components of the fluidics and optics subsystems.
Figure 3-1. BD Accuri C6 Flow Cytometer Optics and Fluidics (Top View, Lid Open)
Set up the computer, monitor, keyboard, and mouse according to manufacturer’s
instructions. Computers come preinstalled with Windows®7 Professional. Contact
Accuri Technical Support for assistance with other operating systems.
Optical
assembly
Fine Particle Filter
Purge Switch
Sheath pump
Waste pump
Red laser
Blue laser
In-line sheath
filter

BD Accuri C6 Flow Cytometer Instrument Manual
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Install the software on the computer before connecting the USB cable to
the cytometer.
NOTE: We recommend making the following changes to the C6 workstation
computer in order to achieve the best possible work environment and to avoid
unexpected software computer issues that could potentially result in a loss of data.
3.2.2 Changing the Workstation Computer Settings for Windows 7
Skip this section if your computer has BD Accuri C6 Software preinstalled.
1. Go to Start (circular icon located in the lower left-hand corner) / select Control
Panel.
2. Select “Appearance and Personalization”
3. Turn off the Sleep Settings.
a. From the column on the left, select “System and Security” then “Power
Options”
b. Select “Change when the computer sleeps”from the left hand margin
(alternatively select “change plan settings” from the main window).
c. From the Edit Plan Settings window, change the “Put the computer to sleep”
setting to “Never” using the drop-down function (we also suggest setting the
“Turn off the display” setting to Never).
d. Select “Save Changes”. Then close the control panel window.
4. Optimize Monitor Display Settings.
(From Start / Control Panel / Appearance and Personalization)
a. Select “Display”, select “Adjust Resolution” from the column on the left.
(Alternatively select “Adjust screen resolution” directly from the Display
section of the main Appearance and Personalization window). Then select
(highlight) Monitor 1.
b. Ensure the Resolution is set to the maximum setting or to at least 1152 x 864.
Change the setting using the slider under the “Resolution” field’s drop down
menu, if necessary.
c. Select “Apply”, then select “OK” and close the Personalization window.
To view hidden file extensions which distinguish BD Accuri C6 Software files (.c6)
from BD Accuri C6 Software template files (.c6t):
1. Click on the Windows Start button and select Control Panel from the pop-up
menu.
2. Select “Tools”from the menu bar.
3. Click on “View”tab
4. Click “Show hidden files and folders”
5. Deselect “Hide extensions for known file types”