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[仪器维护] 您在用什么型号的机器?用于临床还是科研?

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 楼主| 发表于 2013-3-24 13:46:38 | 显示全部楼层
zhangmaodou2000 发表于 2013-3-24 06:55
BD Canto II, 06年左右吧,小毛病数不清,大毛病一次,红光激光的问题,红光报低确实非常常见! ...

我们也是,去年年底刚买的Canto II,最近取的时候已经出现过2-3次blue laser电流检测不到的错误了,每次都是机器外壳的指示灯红灯亮在那里,其它颜色的灯不亮。听他们的工程师说是软件问题,建议关掉软件重连,重启是可以,但我担心如果一直这么下去也不是办法,非常影响效率。
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发表于 2013-3-25 16:34:50 | 显示全部楼层
       看来canto II 红光报警是个常见的毛病,去年让BD的工程师来处理过一次,也就是清洁了一下棱镜,现在又开始报警了,考虑自己动手清洁一下。
    现在科室打算要买台分选机,不知选aria III还是 moflo xDP,有哪位对这两款都了解的,给一下意见!
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 楼主| 发表于 2013-3-25 19:44:02 | 显示全部楼层
mayi009 发表于 2013-3-25 16:34
看来canto II 红光报警是个常见的毛病,去年让BD的工程师来处理过一次,也就是清洁了一下棱镜,现在 ...

国外purdue mail list中在2012年提过一些人对于这两种机型的使用感受和对比,在此转载并稍微总结一下,供您参考,英文不复杂,慢慢看

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.(此人倾向于Aria III,设置和校准时间短)

Jim Phillips
University of Miami School of Medicine
Miami Florida,


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.(此人只谈了Moflo XDP的使用时间经验,Aria没用过,故无法做评述)

Hope the above help.
Bee
Cytometry Core Facility
The Wellcome Trust Sanger Institute
Wellcome Trust Genome Campus
Hinxton
Cambridge
CB10 1SA
United Kingdom


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.(此人亦认为Aria III设置和校准时间短,如果有专门的人员,还是可以考虑Moflo XDP)
Good luck,
Chris


I have a MoFlo and it takes 5 mins to do all laser alignments.(此人居然认为Moflo只需要5分钟进行校准,跟前面几个人相差甚远)
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

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.
(此人认为Moflo和Influx十分稳定,前者12年没出过故障,后者2年,但由于没用过Aria III,故没提及)
--
Jan Bayer, Ph.D.

Flow Cytometry
CEA-DSV-IRCM
Tel : 01 46 54 97 51


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.
(此人是Moflo XDP用户,认为无需2-3小时校准,效率很好)
Regards
Dr.Priya Ramanathan
Lecturer, Dept of Molecular Oncology
Cancer Institute (WIA)
Adyar, Chennai
India

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.
(此人认为,如果你注重用户体验和启动、关机自动化,建议考虑Aria;如果注重仪器扩展和开放性,则可以考虑Influx和Moflo)

Guy Hermans
Principal Scientist
Ablynx nv

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.
(Moflo用户,11年老机型和3年新机型,有时候无需激光校准,即使需要,也只需2-3分钟)
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

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.
(此人有两种机型,XDP只需5分钟进行日常维护,两者均有AUTO-DROP液滴延迟计算能力。Aria更易扩展,但容易宕机。因此该用户倾向于XDP)
Dave

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.(此人认为只有新手才需2-3小时校准Moflo XDP,认为Moflo所需时间并不比Aria III长)

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)
(认为Moflo XDP分选速度更快)
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.
(以上两段讲述了两种机型分选原理上的些许差异,认为Moflo XDP回收率更高)
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.



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发表于 2013-3-26 13:15:25 | 显示全部楼层
呵呵,谢谢啦!我慢慢研究一下!两种机型的价格差不多,就看性能了!
组织样本处理不好?流式中文网原研的魔滤®魔杵®套装,低成本解决,高质量收获
发表于 2013-3-26 17:02:36 | 显示全部楼层
BD FACSCalibur, 07年左右吧,买回后一直闲置大概2年,期间没有专人进行定期维护,造成机器内部管路完全干燥,工程师来处理了一回,后开始正式使用,每周开机使用2-3次,没有出现过什么问题,12年的时候,苹果电脑老是散热不好,系统老死机,不过Calibur没有什么问题。
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 楼主| 发表于 2013-3-26 18:10:51 来自手机 | 显示全部楼层
leiming319 发表于 2013-03-26 17:02:36
BD FACSCalibur, 07年左右吧,买回后一直闲置大概2年,期间没有专人进行定期维护,造成机器内部管路完全干

我们这台04年的,配套的苹果机很少死机啊,你们的苹果是不是后来刚好过渡阶段的cpu?
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发表于 2013-3-27 09:01:02 | 显示全部楼层

嗯,这个不是很清楚了,我进实验室的时候这机子就已经在那里了,苹果系统CPU也没有专门去研究过,不过好像是散热不好的问题吧,我们做流式的时候,整个过程都是在拆开电脑机箱,拿风扇直接对着吹的,这样一直吹着,吹到位了就不会死机(机箱里的风扇都能正常转动,可能风力不够吧),有联系BD工程师,不过要我们自己把主机寄过去,但是,现在已经毕业了,也不知道这个问题得到解决没?
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发表于 2013-4-19 11:10:25 | 显示全部楼层
我们用的是BD VantageSE
大概是2005年左右买的,我是2006年参加工作接手这台仪器
配备了488nm和紫外,但是紫外的激光坏的,一直没有用,所以实际上只有488nm
使用感受:最初接手时,总是为手动校准管路感到很烦,不过后来熟悉之后也还行,及其整体来时运行比较稳定,每周开机3次,每次5小时左右。

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发表于 2013-4-25 11:21:40 | 显示全部楼层
1,BD FACS Calibur ,双激光四色,488和633nm。
2 Aira2 三激光9色。
使用感受:感觉简单的科研检测分析calibur就够用了,复杂的设计用aria2 来做方便些。aria2使用起来不是太方便,总是会有大大小小的问题,点背的孩子伤不起。

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发表于 2013-4-26 09:38:04 | 显示全部楼层
我来说一个款型,Stratedigm的S1000EX系列,三激光十色流式细胞仪主机。十色荧光通道,包括 FITC, PE, PerCP, PE-Cy7, APC, APC-Cy7, Alexa 700, Pacific Blue, Qdot605, PacOr
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