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FlowJo Announcements From: Adam Treister (adam@treestar.com) Date: Mon Apr 22 2002 - 00:15:07 EST • Next message: b cotleur: "2-me in culture media:summary" • Previous message: Geert Raes: "Re: b-mercaptoethanol in media" • Next in thread: Mario Roederer: "Job Opening -- Immediate -- Vaccine Research Center" • Reply: Mario Roederer: "Job Opening -- Immediate -- Vaccine Research Center" • Reply: Michael Dustin : "MoFlo vs Vantage" • Messages sorted by: [ date ] [ thread ] [ subject ] [ author ] [ attachment ] ________________________________________ 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 conveniences, and spiffed it up with the new OS X look and feel. What was already the best analysis software in flow cytometry has gotten a whole lot better. At the meeting, Tree Star is proud to be sponsoring the CyberCafe, your link to home and responsibility while you're in carefree San Diego. Check your email, surf the web, download the slides you'll be presenting at ten. A cadre of California companies has contributed to bring in a premier local roaster to satisfy all your latte urges. We hope you’ll all drop by and see what we mean when we say we're committed to Java. The network is going wireless this year. Just pop a 802.11 card in your laptop, and while your neighbor plays solitaire through the keynote, you can be reading e-mail. We're going to open the CyberCafe with the Second Biennial FlowJo Users Group Meeting,. Saturday night May 4 at 8PM. The first user group meeting was cancelled for lack of interest when Dave Novo brought in a case and a half of French wine, so this year we're going to try real hard to assemble to the point where we can see the show of hands on something before we disband in search of alcohol. We'll have a whole bunch of Macs running FlowJo v4 under OS X, and you can bring your own data and get into big arguments about compensation. Be the first to get the newest FlowJo t-shirt. All you Windows fans out there, come by our booth to see FlowJo running on a PC! We’ll be previewing the long awaited Java version of FlowJo. We’re still not ready to release it, but we’ll be giving a peak as to what it is going to do. 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, Inc. ph: 800-366-6045 intl: 1-650-591-2854 fax: 1-650-508-9186 adam@treestar.com www.treestar.com ------------------------------------------------------------------ ________________________________________ •

FlowJo

From: Manfra, Denise (denise.manfra@spcorp.com)
Date: Tue Jun 19 2001 - 09:37:46 EST


        Hi:

 

               I wanted some feedback on the FLOW Jo application. I

currently do all of my analysis with CellQuest. I am wondering if FLOW Jo

may expedite my analysis. However, I do not use the same markers all the

time, and in one experiment I may be running more than one stain with

multiple different antibodies. Basically, very seldom do I run the same set

of markers. How does FLOW Jo work for such conditions?? How about plot

analysis and excel: apparently Excell FLOW Jo puts the data directly into

EXcell?? What has been your experience with FLOW Jo?? Any information would

be useful.

 

        denise

 

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

 This message and any attachments is solely for the intended recipient. If

you are not the intended recipient, disclosure, copying, use, or

distribution of the information included in this message is prohibited --

please immediately and permanently delete this message.



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

Re: FlowJo

From: Adrian Smith (A.Smith@centenary.usyd.edu.AU)
Date: Wed Jun 20 2001 - 01:25:04 EST


>       Hi:

> 

>              I wanted some feedback on the FLOW Jo application. I

>currently do all of my analysis with CellQuest. I am wondering if FLOW Jo

>may expedite my analysis. However, I do not use the same markers all the

>time, and in one experiment I may be running more than one stain with

>multiple different antibodies. Basically, very seldom do I run the same set

>of markers. How does FLOW Jo work for such conditions?? How about plot

>analysis and excel: apparently Excell FLOW Jo puts the data directly into

>EXcell?? What has been your experience with FLOW Jo?? Any information would

>be useful.

> 

>       denise

 

I have been using FlowJo since it was released in 1997. I haven't

used CellQuest since I started using FlowJo.

 

I have analysed thousands of samples with a lot of different stain

combinations and FlowJo has worked very well for this. In FlowJo you

handle multiple stain combinations by grouping all the samples with

the same stain and then applying an analysis to all the samples (and

you can also make sample-specific adjustments very easily).

 

FlowJo can export statistics for each group and then you simply open

them in Excel. This features has saved me a LOT of time.

 

I would highly recommend FlowJo. I could go on about its benefits but

you can search the archives for my previous posts on FlowJo or read

the review I wrote at

<http://www.biocompare.com/prorev.asp?profrevid=49> for more of my

opinions. Better still, download the latest version and ask for a

free trial serial number. Make sure you do the tutorial and then test

it out on your own samples.

 

If you have any specific questions after you have tried it out feel

free to ask.

 

Adrian Smith



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

Re: FlowJo

From: Giovanna Borsellino (gborsel@tin.it)
Date: Thu Jun 21 2001 - 04:26:47 EST


Hi Denise,

 

         I very highly recommend FlowJo!  In our lab we perform

polychromatic flow cytometry routinely: our samples can be stained with up

to 9 different fluorochromes, and we have multiple samples with different

markers which have to be analyzed in each experiment. We have tried several

software packages, and when we found FlowJo we stopped searching. Actually,

now we use FlowJo even for samples stained with 'only'  3 colours, which we

acquire on the FACScan.

The way FlowJo works is slightly different from other flow-cytometry data

analysis software: basically in FlowJo you create "workspaces" where you

store all the information concerning your experiment: samples, gates,

annotations, statistics, compensation matrices, all your graphs, and

anything else you need for your data analysis. Once you set up the

workspace, data analysis is very fast. A workspace can actually be used as

a 'template'  for batch analysis of any subsequent experiment (this makes

life so much easier!!).

 

         The data with any statistic you can possibly wish for can be

transferred to Excel with a few clicks of the mouse: basically you choose

the statistics you're interested in, and FlowJo will create a spreadsheet

for you with all the calculated statistics from the samples you have

selected. The spreadsheet can then be easily exported to Excel or to

another application.

 

         If you are staining with multiple markers, FlowJo will perform

compensation correctly, no matter what the number of your fluorochromes is

(it is virtually impossible to set compensation correctly with the

'eye-meter'  when your samples have been stained for multiple markers).

 

         The one and only  criticism is that you need a Mac to run it. But

after starting to use it we decided it was actually worth to buy a new Mac,

which is now devoted only to data analysis with FlowJo (so we don't have to

fight over the Mac connected to the FACS...).

 

         It doesn't take too long to learn - of course, you have to make a

time investment, but it will really pay off. The Help files are helpful

indeed - it took us a couple of days, and that was it. You can try it for a

couple of months by downloading the software and getting a free serial

number. Go for it.

 

         I will be glad to show you some of the data we have produced

(FlowJo even creates movies!!) if you are interested.

 

Good luck!

 

Giovanna

(a FlowJo enthusiast)

 

 

>>         Hi:

>> 

>>                 I wanted some feedback on the FLOW Jo application. I

>>currently do all of my analysis with CellQuest. I am wondering if FLOW Jo

>>may expedite my analysis. However, I do not use the same markers all the

>>time, and in one experiment I may be running more than one stain with

>>multiple different antibodies. Basically, very seldom do I run the same set

>>of markers. How does FLOW Jo work for such conditions?? How about plot

>>analysis and excel: apparently Excell FLOW Jo puts the data directly into

>>EXcell?? What has been your experience with FLOW Jo?? Any information would

>>be useful.

>> 

>>         denise

 

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

Giovanna Borsellino, M.D., Ph.D.

Neuroimmunology

IRCCS Santa Lucia

Via Ardeatina 354

00179 Roma

Italy

 

Tel. ++39.06.51501521 (office)

        ++39.06.51501552 (lab)

Fax  ++39.06.51501553

 

e-mail: g.borsellino@hsantalucia.it

            gborsel@tin.it



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

 

 

Re: FlowJo

From: janet dow (jldow@unity.ncsu.edu)
Date: Wed Jun 20 2001 - 11:27:00 EST


I have recently started using flowjo and i love it.  You can go to their

web site, www.flowjo.com and you can get a free 60 day trial on the

software.  I totally recomment trying it.

 

Janet Dow

 

 

 

At 10:37 AM -0400 6/19/01, Manfra, Denise wrote:

>       Hi:

> 

>              I wanted some feedback on the FLOW Jo application. I

>currently do all of my analysis with CellQuest. I am wondering if FLOW Jo

>may expedite my analysis. However, I do not use the same markers all the

>time, and in one experiment I may be running more than one stain with

>multiple different antibodies. Basically, very seldom do I run the same set

>of markers. How does FLOW Jo work for such conditions?? How about plot

>analysis and excel: apparently Excell FLOW Jo puts the data directly into

>EXcell?? What has been your experience with FLOW Jo?? Any information would

>be useful.

> 

>       denise

> 

>***************************************************************

> This message and any attachments is solely for the intended recipient. If

>you are not the intended recipient, disclosure, copying, use, or

>distribution of the information included in this message is prohibited --

>please immediately and permanently delete this message.

 

 

Janet Dow

Research Technician and Manager

Flow Cytometry Facility

North Carolina State College of Veterinary Medicine

Room C-314

Raleigh, NC 27606

(919)513-6364



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

Re: FlowJo

From: Maciej Simm (simmmmer@yahoo.com)
Date: Wed Jun 20 2001 - 17:01:17 EST


Denise,

 

Analysis in FlowJo  works faster than other programs with correct

setup and the right template.

 

Regardless of how many markers you use, FlowJo will automatically

detect your panel settings and you can isolate/sort FACS files based

on their staining protocol (or any keyword) for a separate analysis.

 

Once you analyse some files you can generate statistical tables which

can be copied to clipboard and/or opened in excel.

 

For example, if I have an analyzed set of 1,000 samples for oxidative

burst test and want to generate an excel spreadsheet of

name/date/geometric mean/%positive and CV, I would specify those

parameters in my table editor, and with 2 click of a mouse I'll be in

excel ready to print the results.

 

FlowJo is also a great database tool for FACS files - one application

of that is to find normal ranges of rarely used markers. add a years

worth of FCS filies (provided enough RAM), create a group based on

your marker, apply gates to all of them with one click, and .. voila.

 

Plots can also be copied/pasted (as well as exported)  into

Word/Excel/PowerPoint/Canvas etc.

 

Feel free to contact me if you have any other questions,

 

Maciej Simm

Tree Star, Inc.

 

__________________________________________________

Do You Yahoo!?

Get personalized email addresses from Yahoo! Mail

http://personal.mail.yahoo.com/



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

 

 

RE: FlowJo

From: CJett (CJett@Compucyte.com)
Date: Wed Jun 20 2001 - 15:25:00 EST


Most hardely agree with Adrian Smith...Flow Jo is a time saver for SO many

parts of data analysis.  It does take some time to get used to if you've

been working in the Cellquest reality for long..but well worth it!  Just not

having to enter data by hand is reason enuff!  But I do suggest keeping CQ

around...graphically it is a lil simpler to cut and paste from

 

Cj Jett

Scientist: Biomedical Development

Compucyte Corp.

(617)-577-3811

12 Emily St.

Cambridge Ma 02139-4507



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

Re: FlowJo

From: Phil Marder (mr_redram@hotmail.com)
Date: Thu Jun 21 2001 - 14:39:16 EST


When is the PC version of FloJo coming?

Some of us are waiting for this to give it a try.

 

-Phil Marder

 

 

----Original Message Follows----

From: Maciej Simm <simmmmer@yahoo.com>

To: cyto-inbox

Subject: Re: FlowJo

Date: Wed, 20 Jun 2001 15:01:17 -0700 (PDT)

 

 

Denise,

 

Analysis in FlowJo  works faster than other programs with correct

setup and the right template.

 

Regardless of how many markers you use, FlowJo will automatically

detect your panel settings and you can isolate/sort FACS files based

on their staining protocol (or any keyword) for a separate analysis.

 

Once you analyse some files you can generate statistical tables which

can be copied to clipboard and/or opened in excel.

 

For example, if I have an analyzed set of 1,000 samples for oxidative

burst test and want to generate an excel spreadsheet of

name/date/geometric mean/%positive and CV, I would specify those

parameters in my table editor, and with 2 click of a mouse I'll be in

excel ready to print the results.

 

FlowJo is also a great database tool for FACS files - one application

of that is to find normal ranges of rarely used markers. add a years

worth of FCS filies (provided enough RAM), create a group based on

your marker, apply gates to all of them with one click, and .. voila.

 

Plots can also be copied/pasted (as well as exported)  into

Word/Excel/PowerPoint/Canvas etc.

 

Feel free to contact me if you have any other questions,

 

Maciej Simm

Tree Star, Inc.

 

__________________________________________________

Do You Yahoo!?

Get personalized email addresses from Yahoo! Mail

http://personal.mail.yahoo.com/

 

_________________________________________________________________

Get your FREE download of MSN Explorer at http://explorer.msn.com



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

 

Re: FlowJo

From: Adrian Smith (A.Smith@centenary.usyd.edu.AU)
Date: Thu Jun 21 2001 - 16:44:49 EST


At 10:26 AM +0100 21/6/2001, Giovanna Borsellino wrote:

> 

>         The one and only  criticism is that you need a Mac to run it.

> 

 

Some people would say that is a definite advantage :)

 

At 4:25 PM -0400 20/6/2001, CJett wrote:

>  But I do suggest keeping CQ

>around...graphically it is a lil simpler to cut and paste from

 

But plots looks SO much nicer when they come from FlowJo...

 

 

Adrian Smith



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

Re: FlowJo

From: Carmen Raventos-Suarez (carmen.raventossuarez@bms.com)
Date: Thu Jun 21 2001 - 16:49:26 EST


Denise and everybody else interested in Flow Jo:

I have been using Flow Jo for a year and my requirements are very much like

yours. I also do a lot of DNA analysis, apoptosis and proliferation assays.

We just use the Fl-1,  Fl-2,  etc. as defaults. That way we can run different

set of samples using several different markers with the same dye at a single

run. Later in the analysis we add to the plots the specific name of the

markers. It is fast and efficient, I definitely love it, my analysis time has

been cut down in an exponential way. You create a template and can drop buckets

of samples in it to be analyzed in no time. This includes time for you to view

every single plot in a movie sequence to be sure that all your gates are

correct. Still  if you have cells from different origins and need to correct

gates, this can be done for the individual samples that require correction

leaving the rest untouchable. And the results on excel are at your finger tips

customized by your needs in a couple of clicks. I will never forget all the

macros I needed to built to create individual excel tables from cell quest.

I fewer words, as I told BD who where the ones who introduce me to Flow Jo:

"Flow Jo is like to live in the future and there is no way you want to return

to the past".

Carmen

 

"Manfra, Denise" wrote:

 

>         Hi:

> 

>                 I wanted some feedback on the FLOW Jo application. I

> currently do all of my analysis with CellQuest. I am wondering if FLOW Jo

> may expedite my analysis. However, I do not use the same markers all the

> time, and in one experiment I may be running more than one stain with

> multiple different antibodies. Basically, very seldom do I run the same set

> of markers. How does FLOW Jo work for such conditions?? How about plot

> analysis and excel: apparently Excell FLOW Jo puts the data directly into

> EXcell?? What has been your experience with FLOW Jo?? Any information would

> be useful.

> 

>         denise

> 

> ***************************************************************

>  This message and any attachments is solely for the intended recipient. If

> you are not the intended recipient, disclosure, copying, use, or

> distribution of the information included in this message is prohibited --

> please immediately and permanently delete this message.

 

 

 




This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

 

 

 

RE: FlowJo

From: Gerstein, Rachel (Rachel.Gerstein@umassmed.edu)
Date: Fri Jun 22 2001 - 09:39:24 EST


I think FlowJo will still expedite your analysis, because its quick to call

up new 2-D plots and impose new gates.  And the quick dump into Excell, or

using FlowJo's own table generator will also save time.  I use FlowJo

exclusively and highly recommend it.  Its worth mentioning that its easy to

learn and I rountinely teach undergraduates to do their own analyses.

 

Good luck,

Rachel

 

=======================================================

Rachel M. Gerstein, Ph.D.

Department of Molecular Genetics and Microbiology

Graduate Program in Immunology/Virology

University of Massachusetts Medical School

55 Lake Avenue North

Worcester, MA 01655-0002

(508) 856-1044

(508) 856-5920 (FAX)

 

 

> ----------

> From:        Manfra, Denise

> Sent:        Tuesday, June 19, 2001 10:37 AM

> To:   Cytometry Mailing List

> Subject:     FlowJo

> 

> 

> 

>       Hi:

> 

>              I wanted some feedback on the FLOW Jo application. I

> currently do all of my analysis with CellQuest. I am wondering if FLOW Jo

> may expedite my analysis. However, I do not use the same markers all the

> time, and in one experiment I may be running more than one stain with

> multiple different antibodies. Basically, very seldom do I run the same

> set

> of markers. How does FLOW Jo work for such conditions?? How about plot

> analysis and excel: apparently Excell FLOW Jo puts the data directly into

> EXcell?? What has been your experience with FLOW Jo?? Any information

> would

> be useful.

> 

>       denise

> 

> ***************************************************************

>  This message and any attachments is solely for the intended recipient. If

> you are not the intended recipient, disclosure, copying, use, or

> distribution of the information included in this message is prohibited --

> please immediately and permanently delete this message.

> 

> 



This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:40:22 EST

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Advertisment Not Allowed its called Collusion said:

RE: commercial announcements

From: Dr. Robert Ashcroft (cytomat@netcore.com.au)

Date: Tue Jan 05 1999 - 02:04:46 EST

• Next message: Cheryl Hardy: "viability"

• Previous message: Nick Acquarola: "MoAb against IgM Fab"

• Maybe in reply to: Bill Throndset: "commercial announcements"

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

________________________________________

Dear Bill,

As an academic with commercial connections, I have found that it works if

you do either of two things:

Invite the interested parties to request you reply with a file attached to

their responding email, or simply send an attached file in the original

mailout.

Most people favour the first, as there are lots of people who resent the

attachment file in the primary mailouts.

In the second case, from the marketing viewpoint... the problem is adding

enough detail (without over-sell) in the List message to motivate the

target persons to request the file, yet not enough to alienate the set of

people who are anti-commercial

Hope this helps

-----Original Message-----

From: Bill Throndset [SMTP:bthrondset@rigelinc.com]

Sent: Thursday, December 31, 1998 1:40 PM

To: Cytometry Mailing List

Subject: commercial announcements

Personally, I usually don't mind the commercial comments, but there have

been a few that seem to have crossed the line.

I would suggest requiring a response in which a company directs readers

to products from their company as a solution to a specific cytometry

related problem to include (parenthetically) a warning such as

"propaganda" or "commercial" or "advertisement" with the subject line of

their response. For example; <bold>Subject: re CD34 staining

(commercial).

</bold>For a message from a company which is not a response for help, but

more directly an advertisement, the subject line of the email would also

include "propaganda," or "commercial" or "advertisement."  In this case,

most of us could happily delete the message before reading it!

Maybe it's just me, but I like the paradox of a salesperson typing

"propaganda" as the subject of a listserver posting.

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

bill throndset

bthrondset@rigel.com

Rigel, Incorporated

408-617-8106

________________________________________

• Next message: Cheryl Hardy: "viability"

• Previous message: Nick Acquarola: "MoAb against IgM Fab"

• Maybe in reply to: Bill Throndset: "commercial announcements"

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

________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:36:51 EST

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/>">http://www.treestar.com/flowjo/platforms/> for some information.

• FlowJo is commercially available through Tree Star

• (<<http://www.treestar.com/flowjo>">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 ]

________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:12 EST

Re: Ratio calculation

From: Tree Star (treestar@professionals.com)

Date: Wed Jan 13 1999 - 19:11:06 EST

• Next message: Robert Walt St. George Fisher: "Thank you (responses to: 4N DNA area, but WIDE!!)"

• Previous message: Peter Lopez: "RE: (Fwd) 'alternative' live/dead stains?"

• In reply to: Joseph Webster: "Ratio calculation"

• Next in thread: Ray Hicks: "Re: Ratio calculation"

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

________________________________________

I haven't done it, but you should be able to fake a compensation matrix

that will add FL1 + FL2, then derive a ratio using that new one.  

We considered and dismissed the generalized parameter calculator from

FlowJo because no one had (previously) expressed the need.  If your

colleague can explain why that helps, and it makes sense for others, then

we'll support it.

There is a problem that FCS files don't have negative numbers in them, so

a general algebraic parser wont work.

Adam

On Wed, 13 Jan 1999, Joseph Webster wrote:

>

> Hi All,

> A slightly curly request.... :~?

>

> A colleague wants the ratio of (FL2 / (FL1 + FL2)) from many flow readings.

> (He explained why, but I didn't understand...)

>

> He wants this from many datafiles, so batch processing probably....

> He wants to export this into some other program such as Excel.....

>

> Several computer packages can produce simple ratios of one parameter

> over another, but someone is always wanting more...

>

> Any ideas?

>

> Another challenge for Ray Hicks, Adam Treister, Joe Trotter Et Al ;~)

> How about the ability to enter a user-defined function or formula to be

> applied to the data?

>

> (The "massaging" possibilities are amazing!-)

>

> Thanks, Joseph.

>

> --

> Joseph Webster

> Flow Cytometry Facility

> Centenary Institute

>

________________________________________

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This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:36:52 EST

Re: Cell Cycle Analysis Software-FlowJo

From: Mario Roederer (roederer@stanford.edu)

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

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________________________________________

>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

________________________________________

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

Re: Data Analysis Software

From: Adam Treister (adam@treestar.com)

Date: Thu Mar 18 1999 - 00:00:08 EST

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________________________________________

----->>>>> On 17-Mar-1999, Ray Hicks wrote:

>   The reason for deconvolution is to create a perfect world

>   where those assignments can be made.  The conspiring spread

>   functions are modelled  and how well the model's output

>   matches the real histogram is a measure of perfection.  But

>   the nature of modelling still doesn't allow you to identify a

>   cell.

>  

>   Jill wanted exact boundaries that she could gate on, I don't

>   see how modelling could give them.  It identify a point where

>   there is a satisfactory tradeoff between contamination from

>   an unwanted compartment and loss of the required one, but

>   would it improve the analysis of her horseshoes where she

>   already has an indicator of S-phase?

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

Doesn't this argument come down to a differentiation between a sort gate and

an analysis gate?  At collection time, you don't want to be making decisions

about an individual cell, but in post hoc analysis, there is potentially

interesting information to be gleaned from looking at the overlap

population, or from subtracting it out.

We've been debating this issue internally.  I think it makes sense

statistically, and am lobbying for it in the next FlowJo.  Mario is the self

imposed guardian of scientific rigor and likes to hold his breath until he

turns blue.  But for me, the role of data analysis software is to enable

exploration and to test potential models, not only to churn existing models.

Mark's message lists valid uses for the capability. Clearly there are

limitations, but this is flow, so what else is new?  The software is there

only to calculate the numbers; it's up to the biologist to make sense of

them.

Adam

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

Adam Treister

adam@treestar.com

650-508-9349

http://www.treestar.com

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

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

Re: Macs, BD and applications & instruments

From: Adam Treister (adam@treestar.com)

Date: Mon Apr 26 1999 - 19:57:36 EST

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________________________________________

----->>>>> On 26-Apr-1999, Ulrik Sprogøe-Jakobsen MD wrote:

>   1. Is there a way to run CELLQuest on the blue G3 computers/

>   imacs without hardware hacking? With the lack of an ordinary

>   ADB port, the dongle won't fit, and CQ cannot run without it.

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

The new G3 has an ADB jack in the back (in addition to the two USB jacks).

I bought a 350 MHz G3 this weekend and set it up today.  After getting quite

upset that the cost of having this very fast machine was that I was going to

have to live with the cute-but-not-very-functional keyboard and mouse, I

just discovered the extra ADB jack and I'm now back to my big gray keyboard

that doesn't put the arrow keys directly under the shift key (where I often

hit them and inadvertantly moved the cursor during typing).  Presumably, one

could hang the CellQuest key back there.  

So while I could give you a slew of reasons to use FlowJo instead of

CellQuest, the lack of hardware protection is not one of them  :)   This is

not the case on the iMac.  It only has the USB option for peripherals.  

Adam

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

Adam Treister

flowjo@treestar.com

http://www.treestar.com/flowjo

ph:  (800) 366-6045

fax: (650) 508-9186

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

________________________________________

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________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:06 EST

Re: mac s/w for flow

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

Date: Wed May 05 1999 - 20:42:23 EST

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________________________________________

>Hi Greg

>

>Students and indeed investigators new to analysing and interpreting

>list-mode data shouldn't dismiss the expertise of experienced operators

>offhand. Gating data is a powerful tool. I would start though  with a simple

>package like Cellquest for the mac. Almost everyone I have talked to  has

>found Flow JO  not very intuitive  and don't care to  use it, myself

>included.I also felt it ran very sluggishly on a Powermac-maybe it does

>better on a G3.

>

>

>

>

>Marcia Woda

>Manager, FACS Shared Instrument Facility

>Univ. of Mass Medical School

>Worcester, Ma.  01655

My experience on these points is quite different...

My personal experience is that FlowJo gating is very intutitive and logical

- but not necessarily if you have been trained on programs that use other

gating logic (eg CellQuest). It can be quite a challenge to move over if

you are used to other schemes, but I'm not sure a person new to flow would

find it so difficult (I'm testing this hypothesis at the moment - we have

two people new to flow is lab - one is learning CellQuest and the other

FlowJo...)

As to the speed of running  - when the files are local (ie on your hard

disk) the total time taken to do a basic analysis of an experiment is very

similar between CellQuest and FlowJo (I tried both in parallel when we

first evaluated FlowJo). To me FlowJo certainly didn't feel anymore

"sluggish" on the 7300/180 I was using (although this was with an old

version - the new version of FlowJo has a new live layout editor that can

slow things down, but you don't have to use it, it is just new

functionality that is useful if you have the machine to handle it). When

the files are on the network it is more complicated but most of the time I

think FlowJo would be faster.

In the long term I have found FlowJo an immense time-saver over CellQuest -

all my analyses are recorded and I can go back to them and modify them

easily. I can save stats in a useful form (this alone has saved me hours)

and making pictures for publication or presentation is at least an order of

magnitude better than CellQuest (especially if you don't have room to store

all your files locally and have to fetch them out of an archive).

The other major advantage of FlowJo is you can get a trial version - no

such beast exists for CellQuest (at least not that I know of). You actually

have to buy the package to try it out. If you are planning to do a lot of

flow work think about your analysis package very carefully. CellQuest is

perfectly adequate and does a good job but I personally think FlowJo is a

much better, but obviously some people diasgree. (there was one lab here

that tried FlowJo and then sent it back because they didn't like it -

mainly because it didn't have some abilities they needed - it now has those

abilities).

Good Luck,

Adrian

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________________________________________

Re: formula log to linear??

From: Mario Roederer (Roederer@drmr.com)

Date: Wed Jun 30 1999 - 12:55:10 EST

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________________________________________

>Hello to All,

>

>There is a simple formula for converting log data to linear, and I have

>forgotten.  If anyone remembers this I would appreciate a quick note.

>Thanks.

>

>Jim Phillips

>University of Miami School  of Medicine

>Miami, Fla.

Scale (linear) Value = f * 10^ [(C * n) / R]

where

C = channel number

n = number of decades for the full range (~4 on BD & Cytomation machines,

~3 on many Coulters)

R = range (maximum number of channels; typically 1024)

f = log offset (typically 1 or 0.1)

The FCS keyword $PxE (x = parameter number) has the values for f and n

(e.g., <$P4E = 4,0.1>  means that parameter 4 has a 4 decade range with the

linear scale value of channel 0 equal to 0.1); $PxR has the value for R.

Note that for most BD-generated FCS files, the offset for all log

parameters is incorrectly stored in the FCS keyword list as 0 (zero); it

should be 1.0.

mr

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This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:14 EST

Re: DNA analysis softwares

From: Adam Treister (adam@treestar.com)

Date: Thu Jun 10 1999 - 22:43:01 EST

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________________________________________

----->>> On 10-Jun-1999, Ng Bee Ling wrote:

>   I am looking for a software so as to determine the  % of cell cycle

>   phases (with gating properties) from listmode files. Is there such

>   software that could run both for files collectd from EXPO and

>   Cellquest?

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

FlowJo in an offline analysis package that will read both EXPO (PC) and

CellQuest (Mac) files.   You can compute the percentages in each of the

phases (as well as under-G1 and over-G2) for any gated population.

We currently have a limitation in the ability to export all of the cell

cycle statistics for all of the samples to a spreadsheet in a single step,

but that is implemented in the next release.

More info at:

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

I get a lot of requests for the references to the models, so let me give

them here:

1) Watson Pragmatic

Cytometry 8:1-8 (1987)

Watson, Chambers, & Smith:  A Pragmatic Approach to the Analysis of DNA

Histograms with a Definable G1 Peak

2) Dean Jett Fox

Cytometry 1:71-80 (1980)

Fox:  A Model for the Computer Analysis of Synchronous DNA Distributions

Obtained by Flow Cytometry

3) Two Populations

Uses Dean-Jett model (with Fox modification) for both populations.

Adam

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

Adam Treister

adam@treestar.com

http://www.treestar.com/flowjo

800-366-6045

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

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________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:11 EST

Re: Cost-benefit ratio of FlowJo [Clarification]

From: Adam Treister (adam@treestar.com)

Date: Wed Aug 04 1999 - 16:28:49 EST

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________________________________________

If I may clarify...

Judging from the phone calls I'm getting, Cariappa may

have implied that FlowJo is a cross-platform package.

I think he meant that FlowJo reads data collected on any

cytometer, whether it was hooked to HP, PC or Mac.  FlowJo

itself still runs only on the Mac.  

This may be unclear, because we are running a Summer

$99 promotion, in conjunction with DeNovo Software.  They

sell the PC based analysis package: FCS Express.  So, the

solution I can offer you for Windows is at:

  <">http://www.denovosoftware.com/>

The promotion is a good deal on either platform.

We are working on Win-FlowJo, but it's not close to ready.

The bad news: there is no announced release planned for

a Windows product in the foreseeable future.  

The good news:  more for the Mac is coming pretty soon.

Adam

----->>> On  4-Aug-1999, Cariappa Annaiah wrote:

>  

>   I routinely obtain flow data from both Mac and PC based

>   machines and so have some experience using Mac/PC data

>   analysis software. I have always found it a pain to switch

>   from Mac based software to PC based software and vice versa.

>   That pain is history with FlowJo! The cross-platform

>   capability and the ability to do in-depth, multi-tube

>   analysis has made FlowJo an important member of my pantheon

>   of FCM data analysis software. Now that it is being offered

>   at a really low price for a limited time (as a promotion),

>   the cost-benefit ratio for FlowJo becomes very favourable,

>   compared to extant FCM software. Check it out at the

>   following URL: http://www.treestar.com/flowjo/summer99.html

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

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

Adam Treister

adam@treestar.com

http://www.treestar.com/flowjo

800-366-6045

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

________________________________________

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________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:18 EST

New Mac Analysis Software Launch

From: Ray Hicks (rh208@cus.cam.ac.uk)

Date: Thu Aug 26 1999 - 17:59:38 EST

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________________________________________

This is to announce the release of FCSPress, a new program for the analysis

of flow cytometric data on the macintosh.

Its particular strengths are high quality graphs, on-the-page editing and

annotation, intuitive (to me at least ;-) interface, copy and paste of

graphics and text into other programs (at full resolution - not just bitmap

or screen dump), and ease of use.

You can download a thirty day fully working demo from

http://www.fcspress.com

or

http://www.angelfire.com/biz2/rayh

(the ink hasn't had a chance to dry on the www.fcspress.com address, and it

may not be recognised everywhere on the internet yet).

This release is for powermacs only, a "fat" version for non-powermacs is in

the pipeline.

If you have trouble obtaining a demo from the web, you can request a copy

from request@FCSPress.com <mailto:request@FCSPress.com>.

Pricing for perpetual licence, no time limit, valid for all versions 1.x:

Type Initial cost extra copies

quantity

Single 1 $149 (99 pounds) $149/£99

"Departmental" 5 $749 (495 pounds) $99/£60

"Institutional" 20 $2095 (1395 pounds) $45/£30

For further details see the manual on the web page e-mail

sales@FCSPress.com <mailto:Sales@FCSPress.com>.

All orders received before 14 September 1999 receive a 25% discount, as do

cheque-with-order payments in sterling drawn on a UK bank for single user

licences.

Ray

________________________________________

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________________________________________

This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:37:23 EST

6 : Thu Jan 01 2004 - 17:37:22 EST

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