[Cytometry] COPAS (or similar) large particle sorter

Mikhail Roshal mir9079 at med.cornell.edu
Sat Apr 27 12:11:28 EDT 2013


Hi David, 
Did you give your 130 a shot with the adopocytes. I occasionally sort HRS cells which are about the same size and get half way decent yield and viability.  Obviously a bigger nozzle would be very attractive for me as well.

Misha

On Apr 27, 2013, at 7:18 AM, "David Haviland" <davidspurdueemail at gmail.com> wrote:

> I'd be interested in that... I have a client who has asked about sorting intact adipocytes that can range, so I'm told, from 30-50 microns making the 130um for my Aria out of the running.   
> 
> An anonymous tip suggested I might be able to get a break off with an Aria without a nozzle in place.  So in following his suggestion(s), without a nozzle, I slowly brought the Aria down to 6.1 psi and still had a stream.   At that point I was able to get curves in the stream of "want to be droplets" but there were no conditions running up and down both Amp and Freq ranges by which I could get a droplet break off in the window.   
> 
> Any attempts to drop the pressure below 6.1 psi resulted in the stream collapsing and an ensuing mess.   I gave up the ghost after about 2 hours of futzing with it.   I may revisit it if I end up with a free afternoon.   I have inquired with BD about a 150 or 200 um nozzle but requests have gone unanswered. 
> 
> David Haviland, PhD
> MHRI Houston
> Sent from my iPad
> 
> On Apr 19, 2013, at 8:18, Rachael Walker <Rachael.Walker at babraham.ac.uk> wrote:
> 
>> Hi
>> 
>> By coincidence to this thread, I was told this morning by a scientist I was showing around the lab, that there are modules available to add onto existing sorters to sort large particles such as c.elegans.  Does anyone know if there are such modules available?
>> 
>> I thought on a conventional sorter you can increase a nozzle size up to 200um and if want to sort anything larger you need a Copas.
>> 
>> Thanks
>> 
>> Rachael
>> 
>> 
>> Sent from my iPad
>> 
>> On 19 Apr 2013, at 14:04, "Pulak, Rock" <rpulak at unionbio.com> wrote:
>> 
>>> Hi Rosemary and Vanta,
>>> 
>>> The Aria is OK for eggs/embryos and the first larval stage. Anything bigger (second, third, fourth and Adults stages) starts to become too big for the 100 micron nozzle and the generation of droplets for sorting. Also, there are a lot of other particles, (for example, the cuticles of molts) that can clog the small 100 micron flow cell. Despite these limitations, it does work well for that one early slice of the animal's life cycle. The larger flow cells of the COPAS instruments and the method of sorting, overcome these shortcomings.
>>> 
>>> Best regards,
>>> 
>>> Rock Pulak
>>> Union Biometrica, Inc.
>>> 
>>> -----Original Message-----
>>> From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Vanta Jameson
>>> Sent: Wednesday, April 17, 2013 8:55 PM
>>> To: Rosemary Clarke; cytometry at lists.purdue.edu
>>> Subject: Re: [Cytometry] COPAS (or similar) large particle sorter
>>> 
>>> Hi Rosemary
>>> 
>>> I'm not UK based so can't help you there.  I have had a similar enquiry to sort c elegans on my Aria III, and the researcher sent this Nature Protocols reference where they sort on an Aria I using 100um at 20psi, just lowing the drop drive.
>>> 
>>> Fernandez, A. G., Bargmann, B. O. R., Mis, E. K., Edgley, M. L., Birnbaum, K. D., & Piano, F. (2012). High-throughput fluorescence-based isolation of live C. elegans larvae. Nature Protocols, 7(8), 1502-1510. doi:10.1038/nprot.2012.084
>>> 
>>> We have not yet tried it, but it may work for sorting the smallest larval stage nematodes.
>>> 
>>> 
>>> good luck!
>>> 
>>> Vanta
>>> 
>>> 
>>> ________________________________________
>>> From: cytometry-bounces at lists.purdue.edu [cytometry-bounces at lists.purdue.edu] on behalf of Rosemary Clarke [r.z.clarke at dundee.ac.uk]
>>> Sent: Wednesday, 17 April 2013 5:44 PM
>>> To: cytometry at lists.purdue.edu
>>> Subject: [Cytometry] COPAS (or similar) large particle sorter
>>> 
>>> Dear All,
>>> 
>>> I have been asked by a colleague to investigate the availability of large particle sorters here in the UK. Specifically it would be used for sorting nematode worms. If there is anyone on the list who knows of a suitable instrument in the UK please let me know.
>>> 
>>> Thanks
>>> Rosie
>>> 
>>> Dr Rosie Clarke
>>> Flow Cytometry Facility Manager
>>> WTB/MSI
>>> University of Dundee
>>> Dundee
>>> Scotland
>>> UK
>>> DD1 5EH
>>> Tel: (01382) 385817
>>> E-mail: r.z.clarke at dundee.ac.uk
>>> 
>>> The University of Dundee is a registered Scottish Charity, No: SC015096
>>> 
>>> 
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