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Purdue Cytometry virtually No Trash Emails or Access Barred

Re: List reply-to setting.

From: J. Paul Robinson (jpr@flowcyt.cyto.purdue.edu)
Date: Tue Feb 10 1998 - 12:01:03 EST


 "Mark A. Corio" suggests the need for a listserve approach:

 

Here is my reply:

 

The lsit is run from my lab

 

and

 

Steve kelley send me your message.

 

The way our list is run is by intention.

 

It is designed for maximum value and least annoyance.

 

We believe that a message that is to be

sent to the list myst be intentioned to the list.

 

Otherwise individuals communicate directly.

 

We will not be changing this method

as it is the one favored overwhelmingly by the group.

 

There are virtually no trash emails on this group.

 

There is a very good reason for that.

 

If anyone sends trash mail, they are rapidl;y

deleted from the list

 

and

 

their access is barred.

 

This makes a very

strong incentive to keep a high quality of discussion.

 

My experience

with listserves is the rubbish is overwhelming!

 

We have considered

many times the advantages of a listserve ,

 

and the disadvantages far

outweigh them.

 

Since we are one of the older scientific discussion

groups out there, our track-record speaks for itself.

 

 

We do encourage

people to tabulate results of their questions.

 

They invariably do

this and we thank you.

 

While, I appreciate this might generate some email,

 

let's not fill

the box.

 

We are NOT going to change it anyway!!!

 

regards

Paul Robinson

 

 

> From:        

 "Mark A. Corio" <rmi@frontiernet.net>

> To:            Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>

> Subject:       List reply-to setting.

> Date:          Mon, 9 Feb 1998 17:18:29 -0500

 

>

>

> Steve,

> Belonging to several lists, I found something unusual when I joined this

> list....the reply-to setting is to the original author instead of the list.

>  Several others mentioned it recently and I wonder if the list would like

> to have the reply-to set to the list so that more responses appear on-line

> instead of personal conversations. The list traffic goes up a little, but

> the responses are often more valuable with information than the original

> posts. Anyone else feel this would be a good idea? Is there a reason it is

> the way it is, Steve, that we shouldn't change it?

>

> Mark A. Corio

> Rochester MicroSystems, Inc.

> 200 Buell Road, Suite 9

> Rochester, NY  14624

> Tel: (716) 328-5850

> Fax: (716) 328-1144

> e-mail: rmi@frontiernet.net

> http://www.frontiernet.net/~rmi/

> ****** Designing Electronics for Research and Industry ******

>

J.Paul Robinson, Purdue University Cytometry Labs

Professor of Immunopharmacology

robinson@flowcyt.cyto.purdue.edu PH:765-494 6449 FAX:765-494 0517

web http://www.cyto.purdue.edu



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flowcytometry said:

> RE: Report from ISAC Meeting San Diego

> From: Adam Treister (a...@treestar.com)

> Date: Wed May 22 2002 - 01:45:52 EST

> •     Next message: Susan DeMaggio: "Re: Core Manager's Workshop"

> •     Previous message: Simon Watson: "Computer Networks"

> •     Maybe in reply to: J.Paul Robinson: "Report from ISAC Meeting San

> Diego"

> •     Next in thread: Adrian Smith: "Biotinylation reagents"

> •     Reply: Adrian Smith: "Biotinylation reagents"

> •     Messages sorted by: [ date ] [ thread ] [ subject ] [ author ]

> [ attachment ]

> ________________________________________

> > From: J.Paul Robinson [mailto:j...@flowcyt.cyto.purdue.edu]

> > Subject: Report from ISAC Meeting San Diego

> > Colleagues:

> > Hello from sunny California and the ISAC XXI congress. This message comes

> to you

> > from the CYBER CAFE generously provided by Adam Triester of Tree Star, Inc

> of FLOJO land!

> > Adam has made a bank of 12 computers, a wireless network and, lots of

> network cables for

> >  laptops. He has a T1 fast line and is providing FREE access for the

> entire congress.

> > The room is ALWAYS full and is definitley the most popular place in the

> congress.

> > It has nothing to do with the outstanding FREE coffee from Ryan Bros,

> Coffee again

> > generously provided by Adam. This is the best facility provided by any

> vendor ever!.....

> > so long live FLOJO.....and more free coffee and internet access....

> Paul,

> Thanks for the kind words, but I can't take all the credit for the

> CyberCafe.

> Apple generously provided all the Macs.  IT departments around the

> world may

> say that Macs are hard to network, but we put a dozen Macs on the

> Internet

> in under an hour.   Apple sent top of the line G4s with Cinema

> displays, and

> Titanium PowerBooks, as well as a bunch of iMacs.  Imagine what the

> lines

> would have been like if we only had three computers, as we did in

> Montpelier.

> The true highlight of the cafe was the free espresso. Thanks to

> Phoenix Flow

> Systems, Guava Technologies, PROzyme and Becton Dickinson for

> contributing

> to the coffee fund. By the time we got to the coffee I had blown the

> marketing budget on this endeavor, and these companies stepped in to

> make

> sure you had Ryan Brothers coffee instead of the swill we'd have

> gotten from

> the hotel.  I also extend a special thanks to Kevin Becker for

> bringing the

> Mar Dels to the banquet.  The best band of any ISAC I've attended.

> The CyberCafe crew was Jennifer Wilshire, Maciej Simm, Adam Treat and

> Amy

> Hsu. They thought they were getting a leisurely week on the beach, and

> ended

> up working 9 to 9 every day to keep the CyberCafe running.  It was an

> exhausting schedule, and they were tireless in their support of the

> attendees' Internet needs.  It never would have come off without them.

> Sophia Ascani and Alexandra Treister tie-dyed the 350 shirts.  Each

> one is a

> unique work of art, and each was hand-dipped.  Our garage floor has

> the

> stains to prove it.  So wear them proud, and wash them in cold water.

> We've agreed to do it again at ISAC XXII in Montpelier.  I'm just

> thankful

> this congress is only held every second year.  

All this marketing crap

> just

> gets in the way of my programming.

> Au revoir,

> Adam

> ------------------------------------------------------------------

> Adam Treister

> Tree Star, Inc.

> ph: 800-366-6045 intl: 1-650-591-2854 fax: 1-650-508-9186

> a...@treestar.com    www.flowjo.com

> ------------------------------------------------------------------

> ________________________________________

> •     Next message: Susan DeMaggio: "Re: Core Manager's Workshop"

> •     Previous message: Simon Watson: "Computer Networks"

> •     Maybe in reply to: J.Paul Robinson: "Report from ISAC Meeting San

> Diego"

> •     Next in thread: Adrian Smith: "Biotinylation reagents"

> •     Reply: Adrian Smith: "Biotinylation reagents"

> •     Messages sorted by: [ date ] [ thread ] [ subject ] [ author ]

> [ attachment ]

> ________________________________________

> This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 -

> 17:41:43 EST

6) From: Adrian Smith <A.Sm...@centenary.usyd.edu.AU>

I haven't ever used ModFit so I can comment on it directly, but,

FlowJo also has a proliferation platform that allows you fit curves

to CFSE-type proliferation data and generate various stats and gates.

Overview...

http://www.flowjo.com/specproliferation.html

Details...

http://www.flowjo.com/v4/html/proliferation.html

I believe FlowJo's proliferation platform automatically takes care of

irregularities of log amps by including a parameter to the fit which

models this (I sure Mario could supply the mathematical details to

those so inclined). It can also model the contribution of

autofluorescene. Like most FlowJo analyses it all happens easily and

automatically (but you can adjust parameters manually if you need to)

and it is very easy to apply to batches of samples. Exporting to

Excel is also very easy - as already mentioned by Joanna Roberts in

this thread. I haven't quite worked out how to get it fit curves

every time but overall it seems to do pretty good job on my data.

(Disclaimer: - I provided a some input into the way the platform

works and I have been beta testing FlowJo for a while now)

BTW - after a lot of delays FlowJo 4 is coming along really nicely -

especially if you are using Mac OS X.

I like the new comparison

platform a lot (it is not quite there yet but it shows lots of

potential).

If you have a dual processor Mac you should check out the

latest version which has added support for multi-processors.

If you are worried about upgrading to a new dongle they have just

released v3.7 which works with both v3 and v4 dongles,

just in case

you need to go back to version 3 for some reason.

(Disclaimer:

I also  pushed for this -

there are some very conservative people in my lab

who hate software upgrades!).

Note there was another program being developed specifically for

CFSE-modelling

(I think it was going to be called "CFSE Modeller").

However, it looks to have disappeared and I'm not sure what it's

status is now

I never really had the chance to test it out because

the author was overly paranoid about piracy,

ie there was no way to

test it with your own data.

Given that attitude it is not

particularly surprising that is failed to thrive -

which is a pity

because

Phil Hodgkin was involved in developing it

and he has done a

LOT of mathematical modelling of CFSE-data.

From: Adam Treister (a...@treestar.com)

Date: Mon Apr 22 2002 - 00:15:07 EST

* Reply: Mario Roederer: "Job Opening -- Immediate -- Vaccine

______________________________________...

Only two more weeks until ISAC, that biennial bacchanalia of flower

power

and fun! So I hope you'll excuse a bunch of blatantly commercial

announcements to the list, but endulge me this one time and read on.

At the show we'll be releasing FlowJo Version 4. We've got new

platforms

for overlaying and clustering, we've made it work better across the

Internet, added all sorts of new

For those who haven't found it yet, we've unveiled a spanking new

FlowJo

website. Flowjo.com is chuck full of new content, functionality and

spunk.

Automated price quotes, online ordering, a FAQ that will guide you to

new

depths of understanding, and none of that awful yellow on black text.

The

search engine even works. No ads & cookie-free. Check it out.

Specifically, you should check our pricing. Prices are going up on

May 10.

It has been a number of years since we've changed our prices and with

the

development of the OSX and PC versions, it¹s time for a leap. I

guess

there's no such thing as a free launch. Anyway, this may be a great

time to

buy that FlowJo ten pack you've been thinking about. Licenses

purchased

before May 10 are entitled to a year of free upgrades, including the

4.0

release.

Unleash the flower power!

Adam

--

Adam Treister

Tree Star,

Recent FlowJo announcement

From: Steve Kelley (SKEL...@flowcyt.cyto.purdue.edu)

Date: Wed Dec 30 1998 - 06:13:04 EST

* Next message: Steve Kelley: "Possible minor disruption"

* Previous message: Mark A. Corio: "Chemdex no help..."

* Messages sorted by: [ date ] [ thread ] [ subject ] [ author ]

[ attachment ]

From : J. Paul Robinson <j...@flowcyt.cyto.purdue.edu> ... For data

analysis we want to > use a centraly designed templates made in Flow

Jo software. ...

www.cyto.purdue.edu/hmarchiv/2004/0997.htm

Purdue Cytometry Mailing List: Re: Thanks for the suggestions -

[Disclaimer: Yes, I live off FlowJo sales, and that's a blatantly

commercial statement, but it gets technical from here on.] We've

played with spatial 3D ...

www.cyto.purdue.edu/hmarchiv/2006/0939.htm

To:Cytometry Bushnell, Timothy

Date:Mon, 3 Mar 2008 13:03:53 -0500Content-

Colleagues: Is your lab in need of a new flowjo dongle? I am again

coordinating a bulk purchase of flowjo dongles. The academic price is

$1495 a dongle.

In the past, the price breaks start at 3 dongles and

go up from there. If your lab is interested in purchasing a dongle,

please email me directly the name of the PI, an account number to

charge, and the number of dongles you wish to purchase by March

17th. Tim Timothy Bushnell, Ph.D.Research Assistant Professor,

Pediatrics and OncologyCo-Director, URMC Flow Cytometry

FacilityOffice: 585-273-5535Lab: 585-273-1361Cell: 585-690-5157Fax:

585-276-0233http://www.urmc.rochester.edu/Aab/geneped/flow/http://

www.urmc.rochester.edu/wnyfug/

November 30, 2008 10:20 PM
 

flowcytometry said:

Re: exporting to excel (FlowJo)

From: Adrian Smith (A.Smith@centenary.usyd.edu.AU)

Date: Mon Jul 27 1998 - 21:01:51 EST

•Next message: Houston, Jim : "RE: Sample Differential Failure"

•Previous message: Mario Roederer: "Re: How to do compensation in unlysed sample?"

•Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

--------------------------------------------------------------------------------

>Someone was asking about exporting from CellQuest to Excel-

>

>just to add, FYI, that its very easy to export from FLowJo into Excel; in

>fact you can easily export all

>the stats from a large number of samples very quickly.

>

>Rachel M. Gerstein

>

With the added advantage that all the samples in your group are listed down

the page with the stats across - ready for plotting or further

manipulation. No more re-entering the data from each separate sample into a

spreadsheet.  This feature alone has saved me hours since I switched to

FlowJo.

Adrian Smith - another happy FlowJo user :)

******************************************************

Adrian Smith (PhD Student)

T CELL BIOLOGY GROUP

Centenary Institute of Cancer Medicine & Cell Biology

Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.

Ph: 61-2-9565-6197 Fax: 61-2-9565-6103

A.Smith@centenary.usyd.edu.au

******************************************************

--------------------------------------------------------------------------------

•Next message: Houston, Jim : "RE: Sample Differential Failure"

•Previous message: Mario Roederer: "Re: How to do compensation in unlysed sample?"

•Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

--------------------------------------------------------------------------------

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:35:21 EST

November 30, 2008 10:20 PM
 

flowcytometry said:

Re: Cell Cycle Analysis Software

From: Mario Roederer (roederer@stanford.edu)

Date: Fri Feb 12 1999 - 00:05:01 EST

• Next message: Mark A. Miller: "Re: cell line that expreses CD23 Ag?"

• Previous message: lli@PharMingen.com: "Biotech Job Opening"

• In reply to: Chuck Radford: "Cell Cycle Analysis Software"

• Next in thread: bambam: "Re: Cell Cycle Analysis Software"

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

Does anyone know of any good cell cycle analysis software?

We are currently moving from the HP workstation to the

Macintosh workstation,

And

find that Modfit is a very poor software application.

We really liked CellFit for the HP,

but

BD doesn't do Cell Cycle analysis software anymore,

since they can't make any money at it.

We've also heard of

Phoenix flow systems MacCycle.  

We were just wondering if there were any

others on the market besides those two applications.  Any

help is greatly appreciated.  

Thank You.

FlowJo,

originally developed here at Stanford,

now comes equipped with an integrated cell cycle platform.  

It fits both Watson's "Pragmatic" model as

well as the Dean-Jett model

(with the Fox modification).  

Using an interactive interface,

you can put constraints on many of the fitting

parameters, allowing you to model most DNA distributions.  

However, there

is currently no way to model debris background subtraction

(a process which is controversial),

nor can it model cell divisions

(using dyes like PKH-26 or the like).  

In other words, it's not as sophisticated as ModFit, also available for the Mac,

but suffices for probably 90%

of the users out there!

FlowJo integrates this platform into the unique drag-and-drop interface

that lets you apply your customized models to entire experiments in a

matter of seconds, create graphical reports in which you include

statistics, keyword information, and so forth.  You can even create

template analyses, so that subsequent experiments can be analyzed just by

loading the data into the workspace.

For more information, see the FlowJo web site:

www.treestar.com/flowjo

mr

________________________________________

• Next message: Mark A. Miller: "Re: cell line that expreses CD23 Ag?"

• Previous message: lli@PharMingen.com: "Biotech Job Opening"

• In reply to: Chuck Radford: "Cell Cycle Analysis Software"

• Next in thread: bambam: "Re: Cell Cycle Analysis Software"

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:36:56 EST

December 3, 2008 11:54 PM
 

flowcytometry said:

Re: Flow software for Windows?

From: Marco A. Fernández (marcoa@ns.hugtip.scs.es)

Date: Fri Nov 26 1999 - 03:30:16 EST

• Next message: Katja Adolf: "4 colours on FACS Calibur"

• Previous message: Philippe Bourin: "Re: Flow software for Windows?"

• Maybe in reply to: Are P. Normann: "Flow software for Windows?"

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

There are several windows based software for cytometry data.

Try in these sites:

Catalog of free cytometry software (a lot of useful software)

http://www.bio.umass.edu/mcbfacs/flowcat.html

FlowJo (highly recommended)

http://www.treestar.com/flowjo/

WinList

http://www.vsh.com

FCS Express

http://www.denovosoftware.com./

Hope this helps

Marco A. Fernández

Head Cytometry Unit

University Hospital Germans Trias i Pujol

08916 Badalona (Barcelona)

Spain

marcoa@ns.hugtip.scs.es

http://personal.redestb.es/marcoafdez

----------

De: "Are P. Normann" <a.p.normann@labmed.uio.no>

Para: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>

Asunto: Flow software for Windows?

Fecha: juev., 25 novi 1999 13:35

Hello

We use a FACSort and a FACSVantage in our research and rutine use.  For

analyzing we use Cellquest.  I myself works with research in hematology.

I would like to switch to Windows software and want therefore to gather som

information from other groups who have experience in using Windowsbased

software.

Is there anyone who can give us some advice in this respect

With best regards

Are P. Normann, M.D

Institute of immunology (ITI)

National Hospital

N-0027 Oslo

Norway

Tel.: (+ 47) 22 86 85 61

Fax: (+ 47) 22 20 36 93

e-mail: a.p.normann@labmed.uio.no <mailto:a.p.normann@labmed.uio.no>

________________________________________

• Next message: Katja Adolf: "4 colours on FACS Calibur"

• Previous message: Philippe Bourin: "Re: Flow software for Windows?"

• Maybe in reply to: Are P. Normann: "Flow software for Windows?"

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:30 EST

Re: 4 colours on FACS Calibur-FlowJo

From: Mario Roederer (Roederer@drmr.com)

Date: Tue Nov 30 1999 - 14:13:03 EST

• Next message: Nadeem Sheikh: "detecting vaccinia infection"

• Previous message: Bgreig2@AOL.COM: "Re: Calibur & tube spillover on carousel"

• In reply to: Karenrt@AOL.COM: "Re: 4 colours on FACS Calibur"

• Next in thread: John Altman: "Re: 4 colours on FACS Calibur"

• Reply: John Altman: "Re: 4 colours on FACS Calibur"

• Reply: Margaret Tropea: "Re: 4 colours on FACS Calibur"

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

In contrast to the somewhat misleading commercialism of this post,

Cy7PE was NOT specifically formulated to get around this problem (nor

was it developed by Caltag, as seems to be the implication).  Aaron

Kantor and I (in collaboration with Alan Waggoner) developed this

tandem dye at Stanford to provide us with what was then our sixth

FACS color.  It does have the nice advantage that it can be used in

concert with APC on diode lasers with less compensation than Cy5PE.

However, the assertion that it is not excited by the diode laser is

false; Cy7 can be indeed be excited by the diode laser.  The big

reason for the relatively low compensation between FL3 and FL4 is

because the Cy7 emission is shifted about 30 nm to the red from APC,

so it doesn't fall into the APC filter as much.  Of course, all of

these values depend heavily on the precise excitation line and

emission filters that you use.

Other alternatives to four color systems include using Cy5PE together

with Cy7APC (available from PharMingen, and I think from Caltag as

well), Cy5PE together with Cy5.5APC, or Cy5.5PE or Cy5.5PerCP (from

BD) together with APC or Cy7APC.  (i.e., there are lots of

possibilities; the availability of such conjugates will become far

more prevalent as the multi-color instruments infiltrate our ).

I would remind anyone who uses tandems like Cy7PE that nearly every

different conjugate, even purchased from the same company, may

require a different compensation setting.  This makes proper

compensation on the instrument for multiple staining panels nearly

impossible; currently, the best solution is to use software that can

manage multiple different compensation settings assigned to specific

panels (and in this case, it would be completely accurate to say that

"FlowJo (<http://www.treestar.com/flowjo>) was specifically

formulated at Stanford to get around this problem").

mr

At 5:28 PM -0500 11/29/99, Karenrt@AOL.COM wrote:

>There is another alternative to PerCP for 4-color analysis on the

>FACSCalibur. PE-Cy7 is available from Caltag Laboratories and was formulated

>specifically to get around this problem. The diode laser on the FACSCalibur

>does excite Cy5, causing compensation problems when using it with APC. Cy7,

>however, is not excited by the red diode. There is very little if any

>compensation between FL3 and FL4 with this conjugate, and about 30% between

>FL2 and FL3, For a list of antibodies conjugated to this fluorochrome, check

>our Website at www.caltag.com.

>

>Karen R. Tamul, MS, MT(ASCP)SI

>Mid-Atlantic Territory Manager

>Caltag Laboratories Inc.

________________________________________

• Next message: Nadeem Sheikh: "detecting vaccinia infection"

• Previous message: Bgreig2@AOL.COM: "Re: Calibur & tube spillover on carousel"

• In reply to: Karenrt@AOL.COM: "Re: 4 colours on FACS Calibur"

• Next in thread: John Altman: "Re: 4 colours on FACS Calibur"

• Reply: John Altman: "Re: 4 colours on FACS Calibur"

• Reply: Margaret Tropea: "Re: 4 colours on FACS Calibur"

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

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December 3, 2008 11:58 PM
 

flowcytometry said:

The "perfect" software ... also does ratios... and calibrated parameters...

From: Mario Roederer (Roederer@Stanford.edu)

Date: Fri Jun 18 1999 - 11:30:12 EST

• Next message: Eric Van Buren: "Re: CellQuest Question"

• Previous message: Moss, Delynn M.: "Mac to PC"

• In reply to: Keri Tate: "looking for the "perfect" software"

• Next in thread: biocytex@biocytex.com: "Re: The "perfect" software ... also does ratios... and calibratedparameters..."

• Reply: biocytex@biocytex.com: "Re: The "perfect" software ... also does ratios... and calibratedparameters..."

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

________________________________________

• text/enriched attachment: stored

• <bold>Keri Tate</bold> asks:

• >>>>

• <excerpt>>We have recently purchased a used Coulter XL Epics (1994, 3

• color).  I am trying to find an efficient

• way to analyze data (particularly in messy spleen cell populations).

• Since we have an older model computer (486-66 DTX) with little memory

• we would like to transfer the data and analyze it on a more powerful

• PC. <bold>  My question is what software would you suggest?</bold>

• Although we are not a clinical lab we still do many of the same

• experiments multiple time and would like to save template layouts.  My

• experience is with CellQuest but recently we have being evaluating

• "WinMDI".  My impression thus far is that although the graphics are

• beautiful and the price is right,  it is tedious to crank through data,

• i.e. there are several steps needed to generate one graph and one must

• do this over and over.  Am I missing something?  What other software

• would be useful to evaluate?

• </excerpt><<<<<<<<

• At Stanford, we designed FlowJo for precisely this purpose:  analyzing

• entire experiments at a time, using "template" analyses.  It's batch

• capabilities are particularly easy to use:  you can have it determine

• which sets of gates & statistics to apply to which samples based on the

• staining panels (and other criteria); it can then generate complete

• graphical or layout reports.  Graphics are publication quality: we've

• gone straight from FlowJo output to manuscript submission (although

• usually we use Canvas or other drawing packages to do combine with

• other graphic outputs).  See <<http://

• www.treestar.com/flowjo/platforms/> for some information.

• FlowJo is commercially available through Tree Star

• (<<http://www.treestar.com/flowjo>), but you can evaluate it free for

• 60 days...  I highly suggest running through the tutorial, which leads

• you through this kind of analysis.

• And.... <bold>Chuck Radford</bold> asks

• >Does anyone out there know how to calculate the ratio

• >of fluorescence to FALS of each cell and collect it

• >as a histogram using CellQuest?  I know that Cicero

• >software is capable of doing that, but I'm not sure

• >if CellQuest can do that.

• FlowJo also computes arbitrary ratios (and can convert log <<-> linear

• if desired).  The derived parameter can be used as any other parameter:

• for displays, gating, statistics, and so on.

• Finally, <bold>Bill Hyun</bold> asked about calibration:

• First you need a calibration standard (there are two ways to go:

• either use calibrated bead sets, available from a variety of

• manufacturers, or use an anti-CD4 with a known F/P ratio and stain

• human PBMC; CD4 T cells have 50,000 molecules of CD4 per cell (25,000

• antibody binding sites)).  Once you have collected a calibration

• standard, either beads or cells, FlowJo will use this information to

• create a derived parameter that expresses binding in terms of absolute

• number of molecules per cell.  See <<http://

• www.treestar.com/flowjo/platforms/calibration.pdf>.

• mr

________________________________________

• Next message: Eric Van Buren: "Re: CellQuest Question"

• Previous message: Moss, Delynn M.: "Mac to PC"

• In reply to: Keri Tate: "looking for the "perfect" software"

• Next in thread: biocytex@biocytex.com: "Re: The "perfect" software ... also does ratios... and calibratedparameters..."

• Reply: biocytex@biocytex.com: "Re: The "perfect" software ... also does ratios... and calibratedparameters..."

• Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ]

________________________________________

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December 4, 2008 12:35 AM

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