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FAQ's
We strive to give you the answers you need when you need them. If
you do not find the information you need here, then please email our
Technical Support group at TechSupport@AccuriCytometers.com or
call
734-994-8000 from 8:30 AM until 6:00 PM Eastern Time.
Topics
List of Q's are functional links that jump to the Q+A as shown
below
Capabilities & Performance
 Q:
Does the C6 Flow Cytometer use a traditional sheath and core fluidic stream?
Q:
Can I perform absolute counts with the C6 Flow Cytometer?
Q:
How do I set the voltage gains?
Q:
Do I need to change settings to collect in log or linear mode?
Q:
Can I change the optical filters in the C6 Flow Cytometer?
Q:
Can I use the FL4 fluorescent detector with the blue laser?
Q:
What type of sample tubes can I use?
Q:
Can I sort cells with a C6 Flow Cytometer?
Software & Data
Q:
Do I need to load CFlow software on the computer?
Q:
Can I use CFlow software on a Mac?
Q:
Can CFlow read data produced on another manufacturer's cytometer?
Q:
Can CFlow data be imported into other software programs?
Operation & Maintenance
Q: How
do I clean my C6 Flow Cytometer?
Q: Are
the cleaning and decontamination fluids toxic?
Q: What
type of sheath fluid should I use?
Q: How
do I know if my C6 Flow Cytometer is clogged?
Q: How
do I clear a clog in my C6 Flow Cytometer?
Q: Do
I need to perform scheduled maintenance on the C6 Flow Cytometer?
Q: How
do I remove the SIP?
Q: How
do I install the SIP?
Q: Do
I need to replace the peristaltic tubing?
Q: Does
the C6 Flow Cytometer require special installation?
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. Accuri offers several optional optical filters
(if you can't find what you need on our products page, contact us
and we will do our best to meet your needs). 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: Yes. The C6 Flow Cytometer offers the maximum flexibility
in detector configuration. Because all detectors are focused on a
single spot, it is possible to gather information from either laser
in any or all four color detectors.
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 (with cap removed)
- Other tubes not larger than 16x78mm
- In addition, the SIP Guard can be removed to accommodate a wider
variety of tubes (note, however, that more care will be required
when inserting and removing tubes to ensure the SIP is not damaged).
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.
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Software & Data
Q: Do I need to load CFlow software on the computer?
A: No, the computer supplied with the
unit has CFlow installed and it is ready to use. You may
also purchase additional copies of CFlow to load on your
own computer(s) running Windows 2000 or 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 2000 and Windows XP, but a Mac-capable version
is under development and will likely be available in 2007.
Q: Can CFlow read data produced on another manufacturer's cytometer?
A: Yes. CFlow can import any FCS 2 or
FCS 3 data file. Note that only Accuri's C6 Flow Cytometer is
able to generate the full 6 decades of data, so other files
will be limited in dynamic range.
Q: Can CFlow data be imported into other software programs?
A: Yes. The Accuri C6 Flow Cytometer produces
data over 6 decades of dynamic range, which are displayed
fully by the CFlow software. Some 3rd party analysis programs,
such as WEASEL, are capable of accepting data with such
a broad dynamic range and can open FCS 3 files exported
from CFlow. Other programs cannot accept Accuri's raw data, but can
open Accuri data that has been compressed during export from CFlow.
For example WinMDI cannot read more than 4 decades of data. If you
wish to view Accuri data in WinMDI,it will be necessary to first
export data using the "Export for WinMDI" option under
the File menu in CFlow.
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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 is capable of automatic
decontamination and cleaning. When you press the power
button to turn off the instrument it will automatically
run Decontamination and Cleaning fluids through the fluidics
lines. The Cleaning fluid will remain in the fluidics lines
until the instrument is turned on again by pressing the
power button. When powered on, the instrument will automatically
flush the fluidics lines with sheath fluid prior to reporting
that it is ready for a sample.
It is important to ensure that the sheath, decontamination and cleaning
tanks do not run dry. The system can automatically alert users when
the tanks are low through a display in the CFlow software with optional sensors.
During the automatic cleaning cycles, the C6 Flow Cytometer will draw fluid from
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 Accuri Cleaning Fluid. This will clean both the inside
and outside of the SIP, and help prevent future clogs. When powering
up, it is best to replace the sample tube with one containing 3-4
mL of DI water or sheath fluid.
If necessary, it is possible to activate either the clean or decontamination
function without turning the C6 Flow Cytometer off. Either or both of
these functions can be initiated by accessing the drop down menu
in CFlow. Selecting either option will instruct the C6 Flow Cytometer
to purge the system with the selected fluid, followed by a sheath
fluid flush. Again, placing a tube of cleaning fluid on the SIP can
help clean the SIP and prevent future clogs resulting from salt crystal
formation.
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 DI water. Water will not affect the cells
during analysis, and it is less likely to leave salt crystal
in the fluidics system than saline. However, if the target
particles or cells are very small (<4 microns),
the instrument may produce better quality data if the sheath fluid
and the sample are osmotically similar; generally this means using
buffered saline as the sheath fluid.
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:
1) Remove the sample tube
2) If material is plugging the tip of the SIP attempt to remove by
wiping gently with a Kimwipe or similar low-lint wipe
3) If that is not successful place on the SIP a tube containing DI
water or cleaning fluid
4) Click "Unclog" in CFlow; the C6 Flow Cytometer will then run for about
20 seconds
5) When the cycle is complete remove the tube
6) Click "Backflush" in CFlow; the C6 Flow Cytometer will then run for
about 20 seconds. Visually confirm that sheath fluid is dripping
from the SIP
7) The system is now ready to collect; repeat as necessary
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
- Peristaltic pump tubing elements
The frequency of replacement depends on usage. We recommend replacement
of filters every 6 months, and replacement of the peristaltic tubing
every 12 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: How do I remove the SIP?
A: The SIP Assembly (SIP and SIP Guard)
can be removed without tools for cleaning or replacement:
- If a sample tube is present remove it from the SIP Assembly
- Using one hand, push the Sample Tube Shelf back into the
Loading Bay, out of the way of the SIP Assembly.
- Using your other hand, grasp the upper part of the SIP Assembly
(a plastic cylinder about 2.5cm in diameter) and unscrew it counterclockwise
from the cytometer
- The SIP Assembly can then be drawn down and out of the Loading
Bay.
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 12 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.
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