---- External Email: Use caution with attachments, links, or sharing data ----

test-1 to test list in flat file

No abstract is available available for this message. [Read Message ...]

---- External Email: Use caution with attachments, links, or sharing data ----

XJS*C4JDBQADN1.NSBN3*2IDNEN*GTUBE-STANDARD-ANTI-UBE-TEST-EMAIL*C.34X

---- External Email: Use caution with attachments, links, or sharing data ----

File must be Q with the new policy

---- External Email: Use caution with attachments, links, or sharing data ----

Includes file - after increasing the priority of LsofList policy and waiting for sometime.

---- External Email: Use caution with attachments, links, or sharing data ----

includes file

Test-2

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAF7E6.5B23E9F0]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Test-1

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAF7E5.65574300]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

test1

Test-6

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAF7E1.4BC40210]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Test-5

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAF7DE.FE77B7B0]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Just a sample message

With Correct address this time

test from external to listserv-1

Hello This is Ronnie.

whatever body

---- External Email: Use caution with attachments, links, or sharing data ----

whatever body

Final test -1

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAE362.CCDC49E0]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

test delivery from external - 1.1

test from internal - 4

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAE361.34960230]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

test from internal 3

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.

[cid:image001.png@01DAE360.DF03BA60]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

test-external-2

test from external-1

Hello SamAbusara Test,

Complete with Docusign: TestDMARCdocusignToListserv-1.docx has been voided for the following reason:
Envelope has been deleted and was therefore automatically voided

Envelope ID
dfaab1c8-a3ce-4047-982e-4eeb48a3d170

This message was sent to you by Sam Abusara who is using the DocuSign Electronic Signature Service. If you would rather not receive email from this sender you may contact the sender with your request. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Hello Please sign this document at your convenient.
Thank you

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=8358a3f9-27d3-496e-9594-db5d782308f0&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=2a56f6a9-761f-4b86-93fc-5d3e5a9b86c1

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Hello Please sign this document at your convenient.
Thank you

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=965d615b-4b63-4fcb-b3e9-445b94bf467a&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=2a56f6a9-761f-4b86-93fc-5d3e5a9b86c1

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Hello Please sign this document at your convenient.
Thank you

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=91d55394-e693-414d-a63d-9f83863aa28a&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=2a56f6a9-761f-4b86-93fc-5d3e5a9b86c1

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Hello Please sign this document at your convenient.
Thank you

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=160f6a6e-5ab7-4f35-bd11-d3868f053adb&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=2a56f6a9-761f-4b86-93fc-5d3e5a9b86c1

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Just testing subject tag change

Test after SPF of lists.purdue.edu updated to see if this helps delivery of messages to Yahoo from Listserv.

Ronnie Sarno
IT Applications Administrator
Purdue IT - Purdue Systems Cloud

[log in to unmask]

260-458-4811

[cid:cf25948c-1fa1-4c0e-8e74-3ccd6067989c]

Test-2 Email Check DMARC

Test-1 Email Check DMARC

Hello SamAbusara Test,

Complete with DocuSign: Test Docusign.docx has been voided for the following reason:
Envelope has been deleted and was therefore automatically voided

Envelope ID
a48584f2-35c9-4f83-9e36-af73a287b2a4

This message was sent to you by Sam Abusara who is using the DocuSign Electronic Signature Service. If you would rather not receive email from this sender you may contact the sender with your request. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=a3ae6f60-1bb1-47f4-9a21-9aa471cdd92e&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=17e42266-aa3e-4569-bb41-b1c3ebc76b44&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=1c222177-ec40-4b2c-a686-8b5552930ad9&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=fcebd482-5413-4974-9d62-ee2b918da526&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=022851bc-39ad-4006-be69-2d0a5ce87397&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=155f6f0a-42a9-43b5-ab5b-9b148c34de9e&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=543c48e1-11d2-4fb0-a49c-0fb83df8b861&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=9682370d-ad93-4e33-ae0e-539de783f943&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=b742b44c-7879-45ac-8455-e83638c399a4&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=21658112-be72-49de-b7b6-4cf1f1604df8&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=18963452-599b-4842-8fc0-d5294affd29f&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=d8fb375a-9074-4760-a2f2-45de28f0a216&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=ee53bdff-3581-4c8f-b0d9-fb997ac450f5&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=4787a5bc-9e43-44bd-8c87-8639f7f94488&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Just a Test From Address Rewriting-1

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=13819d3d-f4ef-47d1-9059-7ba3757d215d&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=fd34f9e2-60d5-49d1-ba4a-7515a2db4c54&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=8232fcc8-8af3-4e3c-8f26-abe907f45c42&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Hello SamAbusara Test,

Sam Abusara has sent you a new DocuSign document to view and sign. Please click on the link below to begin signing.

Testing ListServ Behavior message body

REVIEW DOCUMENT

https://na2.docusign.net/Signing/EmailStart.aspx?a=5035d5e3-8e1e-4fce-86a7-c9a088e5924a&acct=a21e2ba6-5ea9-4921-912e-81cbfdb7d154&er=51c1b438-4a77-4632-b491-1f7bd6ebc758

If clicking the link does not work, you can highlight and copy the entire line above and paste it into your browser to get started. 

[More ...]

Test-Send-SubjectTag-01

Test

test

Just a test

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.
[cid:image001.png@01DA60C4.9261CB30]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.
[cid:image001.png@01DA59CE.605D6740]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.
[cid:image001.png@01DA59CD.D9368E90]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Test2

test2

test1

test 2

test6

test5

test4

test3

test2

test1

TEST CHECK AFTER ENABLE CHAGE-LOG

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.
[cid:image001.png@01DA3E4D.9510EFD0]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Test post 2

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.
[cid:image001.png@01DA3E41.21DD7990]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Test1

Sam AbuSara

Productivity Systems Administrator
Purdue IT Messaging.
[cid:image001.png@01DA3D9E.442AE580]
305 N. University St., West Lafayette, IN 47907
[Image result for free fax symbol for email signature][log in to unmask]<mailto:[log in to unmask]>

Agree with everything that has been said! Here are my two cents:

1. It's a great time to plug FixYourFlowDepot, an initiative started by David Leclerc at the University of Chicago to create a video repository on instrument service and repairs. Check it out at https:[log in to unmask]

When we lack the knowledge to fix and repair our own instruments, we are at the mercy of service contracts. 

[More ...]

I was a director of a core flow research lab at a med school. I am long retired but I still have really strong feelings about service contracts. So I am going to vent those strong feelings ? and I am going to try to avoid profanity.

I have no knowledge of the ways that manufacturers model and predict costs and income. I do assume that the end goal of all manufacturers is to make a profit. So the cost of a service contract must, in some way, be set by a prediction of how much can be charged for 

[More ...]

The purpose and structure of service has changed in the time since I left service in 2001. We used to be ?the bartenders? of the industry, trusted by users, enough to talk to pretty openly about their issues and the state of flow cytometry.

Companies started leveraging service in many ways, creating quotas for service contracts that the engineers had to help sell, limiting time able to spend in an account, which caused what I term the ?band-aid? syndrome, only addressing the symptom and not the entire condition of the instrument, and giving engineers much larger territories and many more 

[More ...]

I am definitely in - thanks Vera and Dagna.

Jane

On Tue, Oct 31, 2023 at 12:02?PM Donnenberg, Vera <donnenbergvs at upmc.edu>
wrote:

> Let me think some more?I will have some lab downtime starting next week.
>
> We need to make an outline?.anyone else who wants to participate?
>
> Vera and Dagna
>
>
>
> *From:* Dagna Sheerar <dsheerar at wisc.edu>
> *Sent:* Tuesday, October 31, 2023 1:47 PM
> *To:* Donnenberg, Vera <donnenbergvs at upmc.edu>
> *Cc:* Jane Srivastava <jane.srivastava at gladstone.ucsf.edu>;
> cytometry at lists.purdue.edu
> *Subject:* Re: [Cytometry] An open letter to all 

[More ...]

Let me think some more?I will have some lab downtime starting next week.
We need to make an outline?.anyone else who wants to participate?
Vera and Dagna

From: Dagna Sheerar <dsheerar at wisc.edu>
Sent: Tuesday, October 31, 2023 1:47 PM
To: Donnenberg, Vera <donnenbergvs at upmc.edu>
Cc: Jane Srivastava <jane.srivastava at gladstone.ucsf.edu>; cytometry at lists.purdue.edu
Subject: Re: [Cytometry] An open letter to all companies that provide service contracts (as I know you read this list): 

[More ...]

Vera, I love this idea and would be happy to participate.

Here?s something I?ve been thinking about for a while...

?Cores as Research Infrastructure? to combat the trickle down economics of requiring new investigator recruits to negotiate more money in startup funds to pay for cores and bridge funding to PI?s to pay for cores. Support the corse and the individual research dollars go further. A good argument for both decreasing service contract costs and supporting cores so they can have the cutting-edge high-end instrumentation AND expert staff. 

[More ...]

Dear Jane,

I too thank you for your succinct and right -on summary of issues we all face. As an academic scientist my grants get cuts, not annual increases to cover rate-hikes. The current practice that everything has to "pay for itself" is killing academic science and has to stop.
I am so fed up by the current environment that I am ready to "march for science and research." 

[More ...]

Well said, Jane. Thank you for taking the time to compose and post this. I strongly agree.

Please, I need to unsubscribe. Could you please, help me?

Thanking in advance.

Regards

Does anyone have experience using this antibody in MRD protocol for myeloma
including intracellular starling for kappa and lambda expression ?

thank you

Dear all,

Hope you are doing well! We are in the process of renewing the service
agreement on our BC Astrios cell sorter and wondering if there are any
facilities that have Astrios sorters with "ST Total" and/or "PT Protective"
coverages? If so, would you mind sharing the following information:

1) What configuration of Astrios (in terms of lasers and no. of detectors)
you have?
2) Do you have a multi-year agreement or yearly?
3) How much do you currently pay for the service per year?
4) Typically, what percentage increase in the cost you see during service
renewals?
5) 

[More ...]

I am writing as the core director of a small non-profit research institute
and someone who has been in flow cytometry for quite some time. I know I am
not alone in what I am about to say, and I know that many other core staff
are feeling isolated and caught in the middle of you and the places they
work for. 

[More ...]

Point well taken, Mike... when I do this, there is but a half to one ml
tops in the tube.

The other issue I have to stress with users is not to keep the same tube on
the machine that was just used for priming. It's like blowing your nose,
and then trying to breathe without changing the Kleenex! I know, gross,
but it lends to the point. 

[More ...]

At the risk of triggering a discussion on par with whether "FACS" is purely
sorting or flow cytometry in general, I never "prime" with a tube loaded
(David said to load a tube of water and then prime)--if its too full, then
the air coming out of the SIT will push the liquid up the tube and clog the
line that pressurizes the tube, and I usually want the air to be able to
flow through the SIT without the tube blocking it... Priming while
acquiring is a trick I also use, and will trigger events as the air bubbles
 

[More ...]

On the older Diva Systems you could reboot the internal computer with Tera Term, on the newer systems you can hook a video cable and mouse to the internal Linux box ad see if it is booting. As David mentioned, failure of the internal lithium battery will cause issues. We had this happen on our A5 and the computer would not boot because the time had defaulted to some past time and it caused an error. We were able to connect a keyboard and reset the time so we could start up, but had to do this nearly every time 

[More ...]

Hello Fellow Flow-ers,

I am curious whether anyone has recommendations for 40-60um filter plates or filter inserts for 96-well plates (U-bottom or V-bottom) that can then immediately be used on an HTS system?

Our predicament is that we either produce a significant amount of plastic consumable waste from transferring from staining plate to filter plate and back to a plate that's adequate for HTS or filter each sample individually and then transfer to 96-well plates for HTS use. This laborious process has caused many users to "take their chances" and not filter at all. They get away with it once 

[More ...]

Dear friends and colleagues,

I have two positions available here at the University of Modena, Italy.

Please, spread the word as much as you can.

I have been granted and projects will be starting soon (within the end of the year).

All the best,

Sara

Hello all,

Does anyone have any recommendations for setting gates to capture rare
populations?

I have a user interested in capturing the CD127 population in CSF and is
using veri cell samples to make the FMOs, as CSF was limited. We are seeing
some variation in signal for the same veri cell sample across runs, which
is likely due to the rarity of the CD127+ population. I can follow up with
an image comparing the veri cell sample plots across four different runs,
if needed for reference (please reach out if interested!). 

[More ...]

Hey y'all,
I'm Jacob Kendrick, the Director of Operations at the Flow Cytomety and Cell Sorting Shared Resource Lab at Hollings Cancer Center in Charleston, SC. We are looking to hire a full-time, senior technician to oversee maintenance, use, and application of our Helios/Hyperion Imaging Mass Cytometry system as well as our forthcoming IonPath MIBIScope. We are a medium-sized but well-equipped core at a NCI-designated cancer center at the Medical University of South Carolina, SC's largest healthcare provider. 

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Hi All,

We just wanted to remind you that Derek Davies? next Fundamentals of Flow Cytometry Training Course will be held on the 30th-31st October 16:00-19:30 GMT on both days.

This course will be run by Aja Rieger, manager of the Flow Core and the Advanced Cell Exploration Core at the University of Alberta. You can check out some of her training materials here. 

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I mean this in the best possible way... I'm assuming you don't get the
connection even after a couple of restarts of both the cytometer and the
computer???

In all my years with Diva - quite often my LSRII, Fortessa, A3, A5, and
Aira's I and II, will wake up on the wrong side of the silicon chip and
never connect. OR - sometimes they will show themselves as connected but
aren't. If I don't get a connection, rebooting the machine and
computers sometimes three times will get it back. If not, I ask the FSE
to check the 2032 

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Ensure the nozzle size and associated sheath pressure are sufficient to accommodate the AGG+ aggregates in intact form and orientation as they travel through the laser intercept.

Prateek 

From: Amir Grau <amirgrau at technion.ac.il>
Sent: Fri, 20 Oct 2023 18:41:37
To: "cytometry at lists.purdue.edu" <cytometry at lists.purdue.edu>
Subject: [Cytometry] Pulse shape analysis results
 iWe couldn't verify the sender of this email.Dear All,
 
One of our researcher is running a Pulse Shape Analysis on her cells, for quite some time.
Lately something has change in the way the results look like. The GFP aggregated protein (AGG+) splits in two, as 

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Dear All,

ISAC is seeking innovative research in the biological, life, and physical sciences for presentation at CYTO 2024 from May 4-8 in Edinburgh, Scotland. https://www.cytoconference.org/

Workshop proposals for CYTO 2024 are requested from researchers, shared laboratory personnel, professors and/or practitioners in all settings to offer expertise to the Cytometry profession and should emphasize learning through highly engaging, interactive instruction, and the exchange of ideas and experiences. 

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Dear all
since the 2 dimensional plot seems to be GFP Width vers GFP High, could
it be simply that this second minor population would be doublets and
multiplets in case you did remove the doublets events in a previous
gate? specially if you run at high pressure and high events number per
s.
All best 

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I'd suggest sorting out the two populations and reanalyze them on the same instrument as well as a different instrument. The reanalysis will either show two clean sorted populations, or reanalysis will look the same as the bimodal starting population.

--Peter

On Oct 20, 2023 9:11 AM, Amir Grau <amirgrau at technion.ac.il> wrote: 

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Dear All,

One of our researcher is running a Pulse Shape Analysis on her cells, for quite some time.
Lately something has change in the way the results look like. The GFP aggregated protein (AGG+) splits in two, as if there are two types of aggregations.
These experiments are routinely imaged by microscopy as well, but the images seems to be the same without a clear change as oppose to the flow results.
The cells are acquired on an AriaIII. CST checked and passed. 8 peak beads look terrific.
Question is whether this phenomenon is technical or biological?
In the 

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Dear All

I would like to draw attention to the ISAC Leadership Development Program that is now open (and the deadline has been extended). I personally have benefited tremendously from this program ( I was an ISAC Scholar 2012-2014 and then moved the newly formed Emerging Leader program 2014-2016) and I would encourage anyone interested in becoming a leader in Cytometry to apply. Details of the program and application can be found below and on the website. https://isac-net.org/page/Leadership 

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Hi All,

Does anyone have any experience with using Area Under the Curve
measurements (available under the Kinetics tab in Flowjo) ? I have an
investigator who is looking at K562 cells that have been labeled with a
primary antibody targeting his protein of interest and is hoping to
ultimately sort cells with low and high 'abundance' of protein. To confirm
that his readout is appropriate, he would like to do a comparison of
western blot and flow data - other people have used MFI for this on flow
data, however we were wondering if we could use the AUC 

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The ISAC outreach subcommittee is launching a survey for the January edition of the SRL newsletter. If possible, please post the following blurb and survey link. If you have any questions, please let me know.

The ISAC SRL Outreach subcommittee has put together a survey aimed at gaining deeper insights into the utilization of billing and scheduling software by SRLs for the purpose of instrument management, staff time allocation, and billing, etc. The survey results will be presented in the January 2024 edition of the ISAC SRL Newsletter. This survey should take 5 minutes to complete. If you are interested 

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Hi Marta,
I?d recommend this paper:

Ordo?ez?Rueda, Diana, et al. "Apoptotic Cell Exclusion and Bias?Free Single?Cell Selection Are Important Quality Control Requirements for Successful Single?Cell Sequencing Applications." Cytometry Part A 97.2 (2020): 156-167.
https://onlinelibrary.wiley.com/doi/pdf/10.1002/cyto.a.23898

As well as demonstrating the rationale for sorting live, singlet events they compare apoptotic and ?healthy? cells for their different transcriptomes and do this in quite a simple manner, though from the perspective of apoptotic cell exclusion and the possible consequence of not accounting for these cells. This is from the ?cores? at EMBL in Heidelberg. 

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Dear cytometry enthusiasts,

Are you interested in learning more about computational tools for high-dimensional cytometry analysis and would you like to get some hands-on R experience? Consider traveling to Ghent, Belgium, right before CYTO2024! We organize a 3-day summer school, which will enable you to start exploring your own data in R, as well as include tutorials in FlowJo, OMIQ and FCSExpress from our sponsors BD and Dotmatics. 

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Dear All,

In collaboration with Applied Cytometry, I have set up several Introductory courses. These are suitable for beginners and will cover the basics of fluorescence and fluorochromes, how a cytometer works, experimental design, sample preparation, data analysis and data presentation. Courses are run virtually over two half-day sessions and are taught by acknowledged experts in the field. With consistent training, attendees will have a robust understanding of the technology, be more confident in running a Flow Cytometry experiment and shorten their time to results. 

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Dear fellows

Our FACS Verse workstation had an mental breakdown this week.

The software starts up but it won't connect to the cytometer. We get this message:

Establishing cytometer connection

We would like to get this unit going again without a service call.

Thank you all, Irasema

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Hi everyone!

We have a user who would like to sort apoptotic cells for RNA sequencing. We have absolutely no experience with this. Does any of you have done this in the past that could give us some tips? Or indicate some good reference papers?

Thank you!

Best wishes,

Marta

Hi Will,

??? Could I also get a copy of the results?

Thanks,
Andor

On 9/26/23 1:39 PM, Will Schott wrote:
>
> Thanks to those who have responded to my survey. I have gathered 29
> responses so far and will leave the survey open until October 1^st . I
> will be sending out the results to those who asked after it has closed.
>
> William Schott
> Senior Manager, Flow Cytometry Service
> The Jackson Laboratory
> Bar Harbor, ME | Farmington, CT
> Phone: ?(207) 288-6192
>
> iPhone: (207) 557-4149
>
> Internal Website
 

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Hi Matilda,

From your video, I believe the issue is the placement of your impactor.
My understanding is that for testing purposes the impactor is meant to sample a ?protected area?. In your case the impactor is placed *in* the sort chamber, that is not a ?protected area?. We have a different sorter with a similar door that does not allow access to the sort block, that we sample 5cm back from the closed sort chamber access door, in front of the ?vent?. The placement of the Cyclex-D cassette was written up for example to be placed in the centre 

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Hello Matilda,

We recently tested our Aria IIu using the older Perfetto protocol (using Glo Germ and an impactor) and also saw contamination with our Whisper set on both high and low, but not when the BSC was on. As the instrument was older (2007), we asked our engineer to check the AMS tubing inside the instrument. Turns out they had small cracks in them that was affecting the vacuum within the sort chamber. He replaced all of the tubing inside and our subsequent testing passed. The Fusion should have something similar inside that connects the AMS system to the 

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Hi Matilda,

We have tested our Aria Fusion and struggled with dragon beads when imaging. We has a lot of autofluorecsent garbage on slides sampled without any beads run on the instrument. It helped to spike a slide from the cassette directly with the dragon bead dilution itself to find an optimal focus, exposure etc at the microscope. While FAIL conditions had notable numbers of beads it was difficult to really say there were absolutely zero beads in the test condition with the "noisy" background plus the variation of the focus with the glue on the cassette slides. We have 

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Dear Flowers

We are seeking a flow cytometry technician to join our lab. Ideally someone who has experience in operating a cell sorter. All our sorters are BD with three Fusions, 2 Arias and an Influx. We also have 12 analysers in the lab of various types.
Please see the job description here:
https://www.seek.com.au/job/70500420?type=standout#sol=8e72434a35552c3b0fe9e0233644ccf4b4843fb2 

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Dear fellow cytometrists,

We set up and performed AMS testing today following the Perfetto et al 2018 method from Cytometry Part A "Novel Impactor and Microsphere-Based Assay Used to Measure Containment of Aerosols Generated in a Flow Cytometer Cell Sorter". This is the method with 1 ?m Dragon green beads and Cyclex-D cassettes which we adapted to our BD FACSAria Fusion. I also made use of the Biosafety Sorting webinar on ISACs webportal. However, we detected beads on our unknown sample with AMS set to low and also when tested on high setting. Our positive control with AMS off also 

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And I apparently closed it a day early, oops. I have now reopened it for today.

William Schott
Senior Manager, Flow Cytometry Service
The Jackson Laboratory
Bar Harbor, ME | Farmington, CT
Phone: (207) 288-6192
iPhone: (207) 557-4149

Internal Website<https://jacksonlaboratory.sharepoint.com/sites/FlowCytometry>

To reach all Flow Staff please email:
ME: flowcytometry at jax.org<mailto:flowcytometry at jax.org>
CT: jgmflow at jax.org<mailto:jgmflow at jax.org>

Date: Monday, October 2, 2023 at 1:42 PM
From: <cytometry-bounces at lists.purdue.edu> on behalf of William Schott <Will.Schott at jax.org>
To: Cytometry Mailing List <cytometry at lists.purdue.edu>
Subject: Re: [Cytometry] [EXTERNAL] Survey to compare SRL rates and Services 

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Hi all,
I have received 53 responses so far and will be closing the survey at midnight EDT. Last chance to add to the stats!

William Schott
Senior Manager, Flow Cytometry Service
The Jackson Laboratory
Bar Harbor, ME | Farmington, CT
Phone: (207) 288-6192
iPhone: (207) 557-4149

Internal Website<https://jacksonlaboratory.sharepoint.com/sites/FlowCytometry>

To reach all Flow Staff please email:
ME: flowcytometry at jax.org<mailto:flowcytometry at jax.org>
CT: jgmflow at jax.org<mailto:jgmflow at jax.org> 

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