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Dear Susan,

Our facility is fortunate in having both the PE Ultraview system and a
DeltaVision as well as a Zeiss LSM 510 META set up for live cell
imaging.  They are all absolutely beautiful systems and to be honest I
would be very loathe to give up any one of them!  We do find that each one
has certain advantages over the other so that your applications will
probably determine the best choice for you.  On the other hand, if your
facility is like mine you get all types of applications so that isn't helpful.
So I will try to sum up a few advantages and disadvantages that we have
found, and if you want to discuss it more feel free to contact me at my
e-mail address.

1)  For thick samples, when imaging at lowish magnifications (x40 or less)
we usually find the LSM 510 works the best.  However you haven't asked
about this so I'm not going to say any more about it...

2)  The DeltaVision gives the best resolution of any system I have ever
used, when looking at thin samples.  (Apologies to all other vendors and to
Jim Pawley - I have no numbers at hand to prove it, it's just my
impression.)  The images are stunning.  So if you have cells that are not
too sensitive, and for which you don't need ultra-rapid imaging, you will
get great images.  However, to get the best images you need to deconvolve
from Z-stacks and so if you are looking at very rapidly moving proteins
they may have moved while you acquire through the stack.  For very fast
imaging of e.g. cytoskeletal re-organization we therefore like the PE
system better.

3)  The DeltaVision has the big advantage of its extremely accurate and
stable stage, and the multiple point visiting function.  One of my major
users has his own live cell imaging microscope elsewhere but still comes
down to use the DV every time (even though he has to pay) simply because he
can make 20 movies in a day instead of one.  This function works
unbelievably well.  I have not tried putting a motorized XY stage on my PE
system so I can't comment as to whether that would work as well.

4)  The DV can cause photobleaching and photoxicity problems during the
collection of the stacks.  And this is where the PE system really comes
into its own.  We have had almost NO problems with this on the PE system,
and cells that have seemed to be too sensitive to undergo live cell imaging
on any other wide-field system, or on the LSM, can be imaged for hours on
the PE with no apparent problem.  I myself have found that using one
YFP-tagged nuclear protein I could only collect 4 time points on the
DeltaVision before it had bleached away, but I could keep imaging for hours
on the spinning disk system.  So if you're looking at high magnification
(63 or 100) and using very sensitive cells/fluorochromes then the PE wins
hands down in my experience.  However, be aware that your exposure times
will be much longer on the PE, so that unless you have an incredibly bright
signal you can't really use it at anything like the frame rate it can
potentially acquire.

5)  The PE system can be used with simultaneous DIC and fluorescence pretty
well.  This is also possible on the DV but I am told by the company that
the prisms do affect the deconvolution.  I hope and believe they are
working on a way to get around this problem but it can be a problem at the
moment.

6)  You can do 4 colour imaging on the DV very nicely.  My current PE
system can either do GFP, Rhod and Cy5 together, OR CFP and YFP together,
but not other combinations because you have to swap the fiber optics.  ( I
could get around this but I would lose light).  But you can get newer
versions that are more versatile.  This is just something to watch out for
when you pick your system!

And last but not least....
7)  The DV has the advantage of being all integrated by one company, whose
service, at least in the USA, is very impressive.  My PE system consists of
a Zeiss microscope, the PE head and lasers etc., and then a load of filter
wheels, cameras, software etc. from Universal Imaging, so you can imagine
the problems when something goes wrong and it's not clear whether it's a
microscope problem, or a software problem etc. etc.  It can take a very
long time to sort out who will fix it!

As for heating the cells, we have used the Warner RC-30 and the Bioptechs
FCS2 with both, and I'm currently waiting for a Solent Scientific chamber
to arrive to enclose my whole PE system (they also make one for the
DV).  But I've been writing for hours now so I think I've bored people
enough already...

Hope this is even a tiny bit helpful,
Best wishes,
Alison



>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Our facility would like to purchase a dedicated live cell imaging system
>within the next year (and maybe even by March 2003). High resolution is very
>important to us so we have been looking closely at the Perkin Elmer
>UltraView spinning disc system. Does anyone have anyother suggestions or
>experiences they's like to share? How do you have your systems set up-
>incubation, temperature control? What criteria should we look for? What do
>people think of Applied Precision's DeltaVision in terms of resolution and
>overall?
>
>
>Thanks!!!
>
>Susan Puhalla
>Microscopy Research Assistant
>MRC Clinical Sciences Centre
>Room 5003
>Hammersmith Hospital
>DuCane Road
>London W12 ONN
>UK
>Telephone: 020 8383 8528
>[log in to unmask]

Dr. Alison J. North
Rockefeller University,
Bio-Imaging Resource Center, Box #209,
1230 York Avenue,
New York,
NY 10021,
USA

Tel. (212) 327 7488
Fax. (212) 327 7489
e-mail: [log in to unmask]