Summary Document

This document is a summary of message from the Purdue University Cytometry Laboratories Cytometry Discussion list.

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HI flowers
I want to thank you about the information you give us on your instrument experiences.
We decide for our facility to buy an In-flux (cytopeia): we went to see it in Seattle and
we were really impressed : it's a small instrument that has great potential and the
Cytopeia team is always working towards improvement.

Here a summary of the answered that I received, I hope can be of help to you as were for
us.
thanks again.

Bye
Daniela
dpapini@cnmc.org
202-884-2308

1)
Our sorter and analyzer were installed recently so we are just beginning
to use them. The sensitivity is excellent. Set-up and alignment are
pretty straight-forward. The acquisition software (Spigot) is just for
acquisition, use FloJo for analysis. The company support we've received
has been very good. We plan to use the system for translational
research leading to cell process for clinical trials. I'll begin a
through assessment of the system in January and can let you know more
after that. Other users I spoke to were all very enthusiastic with the
performance of their instruments.

Kevin Sheehan
Stanford Blood & Marrow Transplant Program

2)
I've been using the Influx for over two years now. It's about the best machine I've used.
I think you will really like the InFlux.
I feel its strongest points include:
1. Ease of use. Unlike the Aria, the Influx doesn't need a dedicated operator. Start-up
for analysis usually only take 15-20 minutes and sorting about 1 hour, including
calibrating the drop delay. Its footprint is very small, unlike the MoFlo (with that
damned big optical bench!) that could fit in a corner of a TC room. We have ours in the
corner of a main lab, and we have not had contamination issues, since the Influx is so
easy to clean.
2. The modular design allows for easy upgrade and maintenance. All of the fluidics are
external, allowing for easy replacement and sterilization.
3. Reliability. We've had very little downtime with the unit. Again, the modularity of
the system allows rapid replacement of parts, especially the fuidics.
4. Cytopeia's technical service has been good. Most of the problems have been due to
*user* error, easily taken care of by a single phone call or email. With the very few
components that have failed, replacements have been quickly sent out, usually easily
replaced by myself.
5. In terms of sorting, it's consistently given very high purity rates. I've used it
mostly for single-cell sorting of 293 and CHO cell lines.
6. Software. This is probably the weakest feature so far. The Spigot software is easy to
use and learn, with the split screen feature showing all sort windows in a hierarchal
fashion, unlike the multiple open sort windows of Summit. It's easy to toggle between all
the control functions, draw gates, etc. The compensation matrix is fine, but I tend not
to need it with our current configuration. Spigot is pretty much only for data
collection, with only a few basic real-time analysis functions, and cannot analyze
existing .fcs files. One must use FlowJo or some other commercial analysis suite.
If you have any more questions, please feel free to contact me by email or by phone .
Randy Armstrong
Medications Development Program
Ernest Gallo Clinic and Research Center
University of California San Francisco
5858 Horton St, Suite 200
Emeryville CA 94608
Phone 510 985 3131
Fax 510 985 3101
email randallab@egcrc.net

3)
Hi,
We use an InFlux and a FACSCalibur. The FACSCalibur is more user
friendly and requires less user care, but is much less powerful and
flexible. It's more a black box instrument. It can't to the speed
sorting that the InFlux can, though I think that BD does offer a high
speed sorter. There are many applications we can only do on the InFlux
as it can more readily be tweaked and changed once you know what you are
doing.
Jennifer Jackson
Flow Cytometry Facility
Institute for Systems Biology
Seattle, WA

4)
Here are the responses I got the last time I asked. Please
let me know what you get.
Thanks,
Julie
Reply 1 - I guess I would start with the bottom line, and that is faced with the
choices we had 6 months ago, we would probably (I make this contingent
partly to cover the fact I'm here speaking for others) do it again. We
probably would have gone MoFlo had the prospects of continued support
and continued development seemed more promising. And Aria, at that
point, seemed amiss with its nozzle problems, operational
restrictions, and general 'black-boxish' aspects.
This said about the alternatives, it should also be said that the
InFlux, at least in terms of its software, is a work in progress. It's
only been recently that we got the new board which allows for full,
unrestricted compensation, and as it stands, compensation is still
cumbersome with having to export single color files into FlowJo,
compensating, and re-importing them back into Spigot.
Optics- and fluidics-wise, and probably electronics-wise, it's solid,
which isn't to say, however, that improvements can't be made. For
example, it's hard to run it at MoFlo-like pressures, say 60 psi, not,
I believe, because the electronics won't handle it, but because the
waste bounces fluid back up onto the plates, which are considerably
longer than those on the MoFlo. I've alleviated most of this problem
by putting a little rim of silicon around the waste receptacle. But
generally, I would say it's an easy instrument to operate, and
impressive in the results it can give. Purities of 99+% are not
unusual, particularly when sorting small cells, and it's quite easy to
then change nozzles and sort large cells at 4 psi with the air purge
they've designed into it.
Reply 2 - Our major uses of the Influx in our lab has been for monitoring GPCR
trafficking, clonal cell line sorting and assay development. I've been
using the InFlux (serial #007) for about 1.5 years on almost a daily
basis. It's the best cytometer I've worked with, incorporating many
improvements over the MoFlo. I'm just a very satisfied customer.
Many InFlux features I feel are superior to other sorters:
It has a very small footprint. It'll fit in a corner!
Moldularity of componements. Easily
External fluidics for easy replacement/maintenance, unlike the BD
sorters which have the octagonal fluidic block. This feature has
appealed to the stem cell centers at Stanford, OSHU and UCSF for use
in a GMP environment. The interior of the sort chamber is stainless
steel, with a germicidal lamp. Easy to clean between uses. Cytopeia
offers a soft HEPA enclosure and bladder-style reservoirs of sterile
sheath buffer.
Very high sort purities with high sort rates.
The PMTs are the most sensitive available on the market.
One appealing feature is its ease of set up and use. The operating
software and data collection is straight forward and well layed
out. The folks at Stanford wanted a sorter than did not require
dedicated, experienced flow operators to use.
It takes less time to set up for a sort than other cytometers.
Very reliable. No problems so far with the actual hardware or software.
All problems (the usual user errors......) have been quickly taken
care of by a simple email or phone call.
Reply 3 - We have two Cytopeia instruments, a sorter and an analyzer. This lab is a core
facility with 3 sorters and 8 analyzers. We purchased a FACSAria
and Influx Sorter 2 months apart in 2003. There is no comparison, when

it comes to user friendliness and reliability. The Influx has performed
well and is inexpensive to maintain. In contrast, the Aria has caused us
nothing but problems and is always the last instrument to get booked.
It's routinely "out of service" and is also very difficult to
decontaminate.

5)
We went to Merck to see one in action (sorting bacteria). We were extremely impressed.
We essentially took it all apart and put it back together again. Any replies you get
would be of great interest to us since we are in the midst of sorter searching. Thanks
in advance.
Elaine Kunze

6)
Hello,
I have used both the EPICS and the FACS Calibur for some pretty creative
bacteria work. Both work fine. Pick your flavor!
Caroline

7)

I just went through the same process this past summer. We wanted a
new sorter. It was down to the FACSAria and the InFlux. Before I go
further, I have to say 3 things...
1) I've had several BD sorters, and frankly I've always really liked
them. They sort really well.
2) I like to be "hands on" on my cytometers, and I was fascinated
bythe InFlux.
3) I don't work for either company. I run a core facility at
Creighton University in Omaha NE and I've been in the field for 20+
years.
I've been around long enough to know that claims of "sorting prowess"
by cytometry companies aren't always what they appear. Many
instruments say they are sorters, but many sort only poorly at best.
Others really sort well. I've learned not to trust the
manufacturers. I test it myself, on my samples.
I flew out to San Jose to do a test sort on a FACSAria, and I flew up
to Seattle to do the same on an InFlux. Both instruments were at the
manufacturers site, so I figured it would be a best case scenario for
the sales people. The Aria sorted as promised ... 30,000/s, great
purity. The InFlux ... not so much. We were only able to sort at
low speed/pressure, and the sort was not very pure at the end.
We bought an Aria which was delivered a month ago, and we're very
happy with it. I've run probably 8 sorts on it during this first
month. Results have been very good. We had some "special needs" and
BD was great in putting together a custom instrument for our needs
(like a UV laser). There are some things I don't like about the Aria
and the Diva software, but there were some things I didn't like about
the InFlux and Spigot too. It's probably a wash in those areas.
I would suggest testing the instruments you're interested in
yourslef. In my opinion, at the end of the day, if you really want a
sorter, and you really want to sort with it, there's only one
option. The FACSAria. Cytomation will be out of business soon, and
the InFlux just isn't there quite yet.
I'd be happy to talk with you further if you'd like to write or
call. I don't want to talk down the InFlux too much, because I
really think it has a lot of potential. If you want more details,
call me.
- Greg Perry
Greg A. Perry, Ph.D.
Technical Director
Flow Cytometry Core Facility
Creighton University
Omaha, NE
(402) 280-1841