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1个Purdue上面关于Moflo XDP和FACS Aria系列的讨论，很长，尽管机型偏旧，可能有部分参考价值：
Dear Members,
I received very informative and helpful responses to my post about comparing FACSARIA III and MoFlo XDP. Once again I'd like to thank for kindly sharings. Also I'd like to share all these responses with all listers. I hope it becomes useful for you too.
Dr. Necat Yilmaz
Mersin University
School of Medicine
Histology and Embryology Dept.
Mersin/Turkey
Hi Necat,
I have been working in or running a flow core lab for the past 27 years.
In our lab we have one Aria I and 2 Aria II and a MoFllo. The Arias get
used on a daily basis.. sorting every day... the MoFlo gets used about once
every 6 months. I am not saying the Moflo is a bad machine.. but the set up
time and the calibration is long, long, long. If you have the Moflo you
will need a dedicated operator just for this machine. The Arias, to set
up for an aseptic sort takes about 1.5 hours. This is to clean the machine
and is done mostly automatically. To set up with out a clean takes about
30 minutes. Having used both machines I would very much recommend the Aria
III.
Jim Phillips
University of Miami School of Medicine
Miami Florida,
Hi Sakir,
We have a Mo-Flo XDP in our core facility which we use almost daily.
It took us 10-15mins to align the machine and in between this time to
let the machine stabilise itself before we set up the drop delay.
Average setting up time: ~45mins.
Haven't got a FACS Aria with us so can't comment.
However, as in all cell sorters please make sure the fluidic system is
air bubble free....as the presence of air bubble can affect your drop
delay setting and thus the purity of the sort.
Hope the above help.
Bee
Cytometry Core Facility
The Wellcome Trust Sanger Institute
Wellcome Trust Genome Campus
Hinxton
Cambridge
CB10 1SA
United Kingdom
Hi Sakir,
Its not a 2-3 hour alignment however, it is not an Aria, bottom line - do
you have a dedicated operator? if yes, then XDP would be worth considering,
if no, then Aria is the go, I'd think about the Influx or wait to see what
is released next week in Germany, but every good sorting facility could use
an Aria, super simple and highly reliable.
Good luck,
Chris
Dear Necat
I have a MoFlo and it takes 5 mins to do all laser alignments.
Best wishes Nigel
Nigel G.A. Miller
Flow Cytometrist,
Immunology Division,Dept of Pathology,
School of Biological Sciences,
University of Cambridge,Tennis Court Rd.
Cambridge,CB2 1QP, U.K.
tel:home 01223 323296,mob 07502135226
MoFlo 01223 765666,FACSCan 01223 339086,Cyan 01223 764863
fax 01223 333875
e mail ngam2 at cam.ac.uk
http://www.pathcam.ac.uk
htpp://www-immuno.path.cam.ac.uk/~immuno/flow/flowindex.html
Dear Necat,
Of course the first vendor will tell you the second vendors' system is no
good. The inverse is probably true also. Basically, the last one to listen
to, when it comes to this kind of decisions, is a vendor.
We have used MoFlo for over twelve years now, and InFlux for two. In my
opinion (stress "my" and "opinion" !!) MoFlo (and InFlux) are extremely
stable and reliable systems. We've had essentially no down-time over the
twelve year period without service contract on our MoFlo (cheap operation)
!!! Both sorters represent the fastest systems, and are therefore
extremely fit to stem (rare) cell sorting. Alignment (yes, every day)
takes us 20 minutes, after which we stably sort for hours at very high
rates (40-45,000 per second). Don't ask the vendors whether their machines
are capable of doing this; they'll probably indicate capacities of around
70,000 cells per second (or more).
We're just very happy with these systems (MoFlo & InFLux), and I think
(stress "I") there's no better for stem cell sorting.
--
Jan Bayer, Ph.D.
Flow Cytometry
CEA-DSV-IRCM
Tel : 01 46 54 97 51
Dear Necat,
We have a 4 laser MoFlo XDP that we purchased last year. Its been working
well for us. Alignment is not a big issue though you have to run a check
everyday just to be sure abt your experimental results which I think is a
good thing. You dont need 2-3 hrs to align the laser or stream. As you start
using you will need just a few minutes to check and if need be align .
Alignment will remain the same unless there is some gross disturbance. The
sorting efficiency is also good. Cell cycle maybe a bit problematic because
its a jet in air system. We have found it to be working well for us.
Hope it clears your doubt about the alignment issue.
Regards
Dr.Priya Ramanathan
Lecturer, Dept of Molecular Oncology
Cancer Institute (WIA)
Adyar, Chennai
India
Hi N.,
I had the same choice a few years ago, and requested demos by BeckMan
Coutler, BD (Aria) and DakoCytomation (MoFlo).
The first one got us running in no time, startup being very automated.
In contrast, the MoFlo was prepped for about half a day before we even got
to the site by their local sale/support person. After a full afternoon of
failing to sort even simple commercial reference bead suspensions, we left
knowing A: the machine was much more open but also much harder to set up and
B: local support was going to be worthless.
Coulter had not bought Cytomation from Dako yet, and could only offer
decades old room-sized dinosaurs. We did not even bother looking into those.
Their local distributor is also pretty hopeless in terms of application and
tech support.
The situation has definitely changed over the last years, with upgraded
MoFlo's and descendants now at BeckmanCoulter, and both next-gen Arias and
Influx/JAZZ at BD.
I'd now recommend going with Aria if you value user friendliness and
hands-off startup/shutdown automation, compare Influx/JAZZ and MoFlo if you
really need instrument/exotic application research grade super-flexibility.
G
Guy Hermans
Principal Scientist
Ablynx nv
Technologiepark 21
Tel +32 (0)9 262 01 36
9052 Zwijnaarde
Fax +32 (0)9 262 00 01
Belgium
Cell +32 (0) 470 101 634
Hello Dr. Yilmaz -
You have been terribly misled. Typical MoFlo startup protocol has the
instrument aligned, and drop delay determined, within 10-30 minutes of
startup
(it depends heavily on user's comfort level with the instrument). The
actual
alignment is quite simple.
Consider that the superior viability/vitality/functionality of cells from a
jet-in-air sorter (not a cuvette) far outweighs the small additional effort
required at startup.
Sincerely,
Andrew Beernink
Flow Cytometry Systems Specialist
Beckman Coulter
(858) 774-0425
Dear Dr. Yilmaz,
I have been running a MoFlo for 11 years - 3 years with the newer XDP. It
does not take 2-3 hours to align it in the morning. In fact, most mornings
it does not need laser alignment at all and when it does, 2-3 minutes is all
it takes - one would call it a tweak, not full-blown alignment. It is an
extremely stable instrument and sorts reliably to greater than 95% purity.
I sort ES cells - mouse and human - routinely at least twice a week. They
require a 100 um tip and a lower pressure, around 25-30 PSI to retain their
viability and they sort best at 4 degrees. If you decide to go with the
Aria III, just make sure that you get the bigger tip and a chiller.
Good luck with your decision!
Julie
Julie Nelson
Flow Cytometry Core Facility
Center for Tropical and Emerging Global Disease
University of Georgia
jnelson at uga.edu
706-542-9474
Not true at all about the MoFlo XDP. Yes it needs to be aligned, but usually
takes less than 5 minutes. Beware of companies selling their instruments by
criticizing the competitor.
Joan
Hi,
In our Flow core we have an older version of the Moflo (and therefore
would be consider much harder to use), and if it true that alignment
needs a person which is used to do laser alignments, I should correct
some assertion:
If the machine is well aligned aligned in the beginning of the week
(it can takes you a little while be never 2 to 3 hours!!) and if the
sheath fluid is prepared in the same way every day, then you just need
small adjustments as every machine, in order to gain the max of the
signal when you run your beads for calibration. So, it is true the
moflo is a little bit tricky to align (you have to be used to do this
kind of alignment before) at the beginning, it just needs few
adjustments when it has been already used in the week.
One advantage of this machine is that when you are ok at the sheath
level, it is very very stable. You can sort all day long. And the sort
is as fast as/or better than other machines.
It will depend on your needs. The moflo, as it names says is a Modular
sorter, it means that you can adapt the machine to you experiment (and
not the contrary as the other machines, except the Influx, which as
the same creator) very easily.
If chose the moflo and need any other advice for this sorter, don't
hesite to contact me directly
Regards,
Aude
Dear Dr. Yilmaz,
Our iCyt Synergy cell sorter
http://www.icyt.com/icyt_products_instruments_sorters_analyzers_synergy.php
also requires a daily alignment after startup - but this only takes ~15-20
minutes max.
There are however several significant advantages to a system with computer
controlled laser alignment capability. I will be happy to share this
information - if this is of interest.
Best Regards,
David Merkel
Manager
Merkel Technologies Ltd
David at merkel.co.il
Dear Dr Yilmaz
I operate the MoFlo XDP. What you have heard is an exaggeration. I am
setting up a three color sort today. The longest it would take me is an
hour, if everything went poorly on a Mon morning. It is not a good idea to
listen to the opposition salesperson. I do recommend though that any sorter
be operated by someone with experience and who does at least two sorts per
week. Both instruments are fine, but sorting is an art as much as a science.
By the way, Turkey is one of my favorite places to visit. Where in
Turkey are you?
Hope this helps.
Barry Udis
We have both XDPs and Arias. While the XDP does require alignment on a
mostly daily basis, it takes all for 5 minutes to perform. Both instruments
have auto-drop delay calculations, and both function in high and low
pressure environments. The Aria is more expandable (in that you can add up
to six independent lasers), but are much more prone to failure. We liked
the XDP so much, that we were willing to invest in the Astrios (the other
sorter from Coulter) when replacing some of our old systems.
Dave
Hi Sakir,
We have and Aria I (the same alignement that in Aria III) and a Moflo (Not
XDP)....and we can say you that Aria is simpler than Moflo. With Aria you
don't need to align every day and maybe Twice a year if all is ok...With
Moflo you need to align every day...or better you need to check the
alignement every day...The question is...Who is in front of the
machine?....If you have a technician dedicated to this then it doesn't
matter wich one you choose in order to their friendly use...The technician
could achieve the best performance in both equipments...Just take care that
the one you choose cover your necessities...Don't hesitate with the user
friendly capabilities if your technician is good. sure the Aria is easy to
use...but that shouldn't be the main reason to choose it...An important
reason...but not the one. If the necessities of your service are covered by
the two machines then...The easy way of Aria could be an advantage..
--------------------------------------------
De idiomes en sabem uns quants...de Llengua només una, el Català.
2012/6/11 Julie Nelson <jnelson at uga.edu>
Dr Yilmaz
Our facility has an upgraded FACSAria-IIIu. It is an AriaI that has been
upgraded to an AriaIII fluidics. We also have a 2 laser XDP that our lab
has had for about 8 months. I would not consider myself an expert on the
Moflo, but am getting better.
I will say that I love the ease of starting the FACSAria-IIIu. If you get
a good nozzle, everything goes very smoothly. Sorts are generally good.
That said, I find that the Aria is more harsh on larger cells such as
Dendritic cells, macrophages, and epithelial tumor lines. I also feel that
sorting into 384 well plates can not happen on the Aria.
We are performing a good deal of single cell sorting into 384 well plates
for single cell gene arrays. We are using our XDP exclusively for this and
getting phenomenal results.
I think that (in general) jet-in-air systems are gentler on cells and more
accurate for sorting. Our XDP operates at a higher frequency, which
translates into faster sorts.
That said, my Moflo XDP is like taking a step back in time with more
manual set-ups. You do have to align the lasers, (every time). Once tuned
int to a specific nozzle (tip), there is very little aligning that needs
to happen, I just verify it. Once you get used to this procedure, it will
take about 1-5 minutes to align. I should say that this was a head ache at
first because I never had to align it in the past. I now don't have to
wait for a service engineer to come out to align them. It is very easy
after you get used to it. The main issue is getting you stream vertical.
This wrote up a step by step (if then) white sheet for our technicians to
follow. Once you get used to it, you can align it fast. IT TAKES
PRACTICE!!!! I will definitely give the XDP a MUCH steeper learning curve.
The intellisortII has automatic frequency find, and beadless drop delay,
both of which I really love (when they work) for the 70um and 100um tips.
I am told that BC is working on a solution to the other nozzle sizes so
that the automatic drop delay will work for other tips. If you want to use
larger tips, you will have to manual do drop delay by counting beads in a
"puddle" using a microscope.
The intellisort is a little buggy, but after spending time beating your
head on the wall and calling technical support, you will find quick tips
around the bugs.
I feel that the fluidics are overall, more stable than my aria. You
really, I mean really need to take care that ALL bubbles are out of the
flow cell assembly. There are some tricks for this to make it quick, but
it was not stressed enough during our training and cause me some issues.
Once fixed, it have had no issues with drift of my frequency (for the most
part).
I personally,like Diva better than summit, but it could be that I've been
running Diva since 2005 and am quite used to it. It is always painful
learning new software and hardware.
My over-all impressions. The Aria will be easier to learn and is a good
instrument. The XDP is slower to learn, but will sort faster and much more
consistent. I also had no one to "hold my hand" for either machine. Early
on for each machine, the service or help line was my best friend.
If you have any other questions, please do not hesitate to ask.
Ben
Benjamin J. Daniel, Ph.D.
Assistant Professor/Research
Dept. of Molecular Medicine
Flow Cytometry Facility Director
Certified Cytometrist
Univ. of Texas HSC at San Antonio
7703 Floyd Curl Drive, Rm 5.044V
Mail Code 7765
San Antonio, TX 78229
Phone (Main Campus): 210-567-3911
Phone (STRF): 210-562-4065
Fax: 210-567-2053
Website: http://research.uthscsa.edu/facs/index.asp
Dear Dr Yilmaz,
Only a blind man could take 2-3 hours to align a MoFlo XDP. I learnt my
flow cytometry on one of the very first MoFlo MLS (No. 10003), it was only 2
laser but I could get that machine from cold start to running a sample in
not much more than it takes to get an Aria III running, including aligning
the lasers.
The Aria does not require laser alignment, but it is a much slower
instrument. Optimaly I use a 85um nozzle at 45psi (~45,000 droplets/sec)
and can sort at a sample rate of 9,000 events/sec. The Aria (I have an Aria
IIu) works best at 1 sorted droplet per 5 droplets created. (You wil find
that despite the hype most Aria users don't use the 70um nozzle or 70psi)
The MoFlo XDP (I have not yet used one but will have one at the end of the
year) is designed for high speed sorting, with a 70um nozzle, at 100,000
droplets/sec (60psi), I have sorted a sample at 34,000 events/sec (MoFlo
MLS).
I currently use my Aria IIu for haemopoeitic stem cell sorting, it is
painfully slow and I must sort at least 20% more cell than the researcher
requires so they are even close to the number they require.
The major difference between the two instruments is when they interrogate
the cell. This is a pressure related problem, before the nozzle the cell is
subjected to sheath pressure plus sample pressure - so on an Aria 45+psi,
after the nozzle it is experiences a sudden pressure drop to atmospheric
pressure which will cause sensitive cells to disintergrate. (E.g. foetal
pig cells are exremely pressure sensitive and have to be sorted at 10-20psi
through a 100um nozzle.)
The Aria looks at the cell when it is still inside the flow cell, it is
claimed this gives better resolution, (therefore you can run lower power
lasers), but if the cell is damaged as it passes through the nozzle the
instrument proceeds to sort the now empty droplet. This companies own white
paper quotes no more than 80% of number of target cells will be found in the
collection tube. My personal experience (depending on cell type) is
recovery can be anywhere between 10 & 85%.
The MoFlo interrogates the cell after it has passed through the nozzle, just
before the droplets are formed. If the cell is damaged passing throught the
nozzle it is not there to be analysed. Therefore recovery is 95-99%. You
never get 100% recovery, some cells always die post sort.
I am probably biased, but I would suggest the MoFlo XDP.
Remember it is a very competative business selling cell sorters, each
company will say their instrument is the best and the rivals is no good.
Other instruments of possible interest are the BD Influx (a MoFlo clone, but
allegedly designed for biohazard hood use), the Beckman Coulter Astrios (in
my mind a much better instrument than the Influx, the Astrios was designed
from the start for Biohazard hood use), and the Sony iCyt sy3200 (this is
new, but offers an instrument purpose designed for a biohazard hood).
Regards
Robert Wadley
Cytometry Manager
Cytometry & Imaging Suite
Mater Medical Research Institute
Brisbane, Australia.
> Date: Sat, 9 Jun 2012 01:49:28 +0300
> From: nyilmaz at mersin.edu.tr
> To: cytometry at lists.purdue.edu
> Subject: [Cytometry] Facsaria III vs moflo xdp
Hi Necat,
Our facility has a Moflo Legacy installed since 2001. It is older than the
XDP version but I can tell you a few things about it.
First of all, none of the systems (Aria or Moflo) are user friendly to
operate!!! Don't believe anyone saying a cell sorter is easy to operate!!
Especially for new operators/flow cytometrists.
The main difference on these systems is the fact that the Moflo needs
alignment everyday at the start-up. The 2-3 hours is grossly
over-evaluated!!! This is a misleading information. All in all, it takes me
30 minutes of preparation time (including laser warmup, filling the tanks
and drop delay determination) every morning. Not much more than the FACaria
III which is alignment free...
Hope that helps!
Eric Massicotte
Responsable, Plateau de cytométrie en flux
Manager, Flow cytometry facility
Institut de Recherches cliniques de Montréal (IRCM)
110 ave. des Pins Ouest
Montréal, Qc
H2W 1R7
Tél. 514-987-5627
It takes about 5-10 minutes to align an XDP. In fact, one of the best
features about an XDP is that you can align it yourself so you don't have to
wait for a repairman.
Karen Helm
University of Colorado Denver
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发表于 2018-1-3 06:23:06
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