FAQ's

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Capabilities & Performance

Q: Does the C6 Flow Cytometer use a traditional sheath and core fluidic stream?

A: Yes and No. The fluidics design of the C6 Flow Cytometer is unique, but it does generate laminar flow with hydrodynamic focusing of sample in a core fluid surrounded by a sheath fluid. However, unlike existing cytometers, with the C6 Flow Cytometer the user can independently control the speed of the core (sample) fluid and the diameter of the core. This differential control makes it possible to set an optimal core diameter for your sample while independently setting an optimal flow rate. This feature increases the precision of sample analysis.

Q: Can I perform absolute counts with the C6 Flow Cytometer?

A: Yes. The C6 Flow Cytometer simultaneously measures events (cells) and the volume of sample fluid passing through the system. From this information CFlow can calculate the absolute number of events per microliter of sample fluid.

Q: How do I set the voltage gains?

A: The C6 Flow Cytometer has a broad dynamic range and does not require the user to manipulate voltage gain settings. All available sample data can be collected with excellent resolution using the factory settings. This feature eliminates a time consuming step in data collection, and greatly reduces the chance that data will be lost.

Q: Do I need to change settings to collect in log or linear mode?

A: No. The C6 Flow Cytometer collects all data in linear mode, and CFlow can display the data in linear or log format. The change from linear to log can be made or reversed at anytime during or after data collection. The default display settings for CFlow display scatter data in linear format and fluorescent data in log format.

Q: Can I change the optical filters in the C6 Flow Cytometer?

A: Yes. The protocol for replacing optical filters is easy and takes less than 5 minutes.

Q: Can I use the FL4 fluorescent detector with the blue laser?

A: No. FL4 is dedicated to the red laser.

Q: What type of sample tubes can I use?

A: The C6 FlowCytometer does not use a pressurized fluidics system, so it is very tolerant of the type and brand of sample tube used. The Sample Tube Shelf can accommodate several types of tubes, including:

12x75mm tubes, any brand
Most microcentrifuge tubes
Other tubes not larger than 16x78mm
In addition, the SIP Guard can be removed to accommodate a wider variety of tubes.

Q: Can I sort cells with a C6 Flow Cytometer?

A: No. The C6 Flow Cytometer is an analytical cytometer. The test sample (cells) is discharged into the waste tank after analysis.

Software & Data

Q: Do I need to load CFlow software on the computer?

A: Yes, the computer supplied with the unit does not have CFlow installed. You may purchase additional copies of CFlow to load on your own computer(s) running Windows XP. CFlow is quick and easy to install.

Q: Can I use CFlow software on a Mac?

A: Not yet. CFlow currently runs on only Windows XP, but a Mac-capable version is under development and will likely be available late in 2008.

Q: Can CFlow read data produced on another manufacturer's cytometer?

A: No. CFlow is designed to only work with data collected on the C6. Howerver, CFlow can export data as FCS 3.0 files which can be compared to data from other manufacturer's cytometers using 3rd party software programs such as FCS Express and FloJo.

Q: Can CFlow data be imported into other software programs?

A: Yes. Even though the Accuri C6 Flow Cytometer produces data over 6 decades of dynamic range, most 3rd party analysis programs, such as FCS Express and FlowJo, are capable of accepting data with such a broad dynamic range and can open the FCS 3.0 files exported from CFlow.

Q: What is the difference between CFlow and CFlow Plus?

A: Two software packages are available for the C6 Flow Cytometer: CFlow and CFlow Plus. CFlow offers all of the cytometer controls and analysis features necessary to run the cytometer, collect data, and analyze results.

CFlow Plus contains all the functionality of CFlow and has the additional advanced features of user-adjustable, independent fine control of core size and flow rate, the ability to measure and record sample volume, and the addition of the time parameter enabling kinetic measurements of all other parameters.

Operation & Maintenance

Q: How do I clean my C6 Flow Cytometer?

A: Generally, all you need to do is turn the instrument off at the end of each day by pressing the power button. The C6 Flow Cytometer will automatically decontaminate itself. When you turn off the instrument it will run decontamination and sheath fluids through the entire fluidic path. When powered on, the instrument will automatically flush the fluidics lines with sheath fluid prior to reporting that it is ready for a sample. In the case of an unexpected loss of power, the C6 will decontaminate itself during the next power up before reporting that it is ready for a sample. Additionally, there is a Cleaning cycle that can be run periodically from the Instrument menu. Accuri recommends running this second cleaner cycle once a week unless the samples are unusually sticky or problematic.

It is possible to activate this decontamination cycle without turning the C6 Flow Cytometer off. The Decontamination Cycle is accessible under the Instrument menu. Selecting this option will instruct the C6 Flow Cytometer to purge the system with the decontamination fluid, followed by a sheath fluid flush.

During the automatic cleaning and decontamination cycles, the C6 Flow Cytometer will push fluid down the sample injection probe (SIP). When shutting down for the day, it is a good idea to place a sample tube on the SIP that contains 2-3 mL of sheath fluid. This will clean both the inside and outside of the SIP, and help prevent future clogs.

It is important to ensure that the sheath, decontamination, and cleaning tanks do not run dry and that the waste tank does not overflow. The system is equipped with fluid sensors in each tank to alert the user when they need attention.

Q: Are the cleaning and decontamination fluids toxic?

A: Accuri's Cleaning fluid contains a biodegradable detergent. Accuri's Decontamination fluid contains reagent-grade bleach. Both can be disposed of down the sink.

Q: What type of sheath fluid should I use?

A: For most cell analysis we recommend using high grade filtered DI water. Water will not affect the cells during analysis, and it is less likely to leave salt residue in the fluidics system than saline.

Regardless of type of fluid used, the instrument will perform better if the sheath fluid is free of debris such as dust or microbial growth.

Q: How do I know if my C6 Flow Cytometer is clogged?

A: Depending on the type and quality of sample used, it is possible for the SIP to clog. The inner diameter of the SIP is 200 microns, so clumps of cells or debris may not be able to enter the fluid path. A clog in the SIP will prevent sample uptake, resulting in a sudden reduction in detectable events. Often a clog can be confirmed by visual inspection of the SIP tip.

Q: How do I clear a clog in my C6 Flow Cytometer?

A: To clear a clog in the SIP:

Remove the sample tube.
Place a tube containing clean DI water on the SIP.
Click "Backflush" in CFlow; the C6 Flow Cytometer will then run for about 30 seconds. Visually confirm that sheath fluid is dripping from the SIP while Backflush is running. If fluid is not dripping from the SIP while Backflush is running, run Backflush several times in succession until fluid can be seen dripping from the SIP.
If fluid does run out of the SIP during Backflush and yet the C6 still appears clogged, click "Unclog" in CFlow; the C6 Flow Cytometer will then run for about 30 seconds. This purges the flow cell.
Check to see if your sample event rate is normal.
The system is now ready to collect.

Q: Do I need to perform scheduled maintenance on the C6 Flow Cytometer?

A: Yes. Turning off the instrument at the end of each day, and cleaning the sample injection probe (SIP) regularly are good first steps (make sure the sheath, decontamination and cleaning tanks do not run dry). There are two parts that need to be replaced over time:

Fluid filters (4)
Peristaltic pump tubing elements (2)

The frequency of replacement depends on usage. We recommend replacement of filters every 2 months, and replacement of the peristaltic tubing every 3 months, or more often if usage exceeds a couple hours per day. The replacement protocol is easy and quick and can be found in the C6 Flow Cytometer Instrument Manual. Replacement part kits are available from Accuri Cytometers.

Q: How do I remove the SIP Guard?

A: The SIP Guard can be removed without tools for cleaning or replacement:

If a sample tube is present remove it from the SIP Stage.
Push the Sample Stage back all the way. It will click in the back position.
Gently pull the Guard forward while simultaneously pulling straight down. Do not twist the SIP Guard to remove it.
Replace by pushing the SIP Guard back up into the black SIP Collar.

Q: How do I install the SIP?

A: To reinstall the SIP Assembly:

Using one hand, push the Sample Tube Shelf back into the Loading Bay, to create space to insert the SIP Assembly.
Using your other hand, carefully place the top of the SIP Assembly into the hole above Sample Tube Shelf. Be careful not to damage the SIP's narrow diameter tip by contacting it with the surrounding plastic or metal.
The SIP Assembly should reach about 1cm into the hole before it abuts the threads in the hole.
When the top of the SIP is in place, screw the SIP into place. Be careful to not force the SIP into place or strip the threads. It should turn easily.
Stop when it feels tight. The SIP seals with a rubber O-ring; it does not need to be screwed in very tight. Screwing in the SIP too hard can crack the plastic part of the SIP and even damage the cytometer.

Q: Do I need to replace the peristaltic tubing?

A: Yes, the peristaltic tubing elements do wear out over time and should be replaced. The frequency of replacement depends on usage. We recommend replacing the peristaltic tubing every 2 months, or more often if usage exceeds a couple hours per day. The replacement protocol is easy and quick, and replacement kits are available from Accuri Cytometers.

Q: Does the C6 Flow Cytometer require special installation?

A: The C6 Flow Cytometer is a bench top instrument, and it operates on a universal power supply (100-240V, 50-60Hz). There are no special electrical requirements for installation.