Documenting Bead Lots in Experiment Descriptions

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John Sexton

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Jan 8, 2017, 10:06:14 PM1/8/17
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Howdy folks. I've been playing around with the mini inter-lab data, and I was unable to find documentation of the bead lots used by David and Ariel in their respective experiments. If you guys could let me know or update your documentations, that would be much appreciated! (sorry if it's there and I missed it!)

(I ultimately need the MEFL and MECY table values for each of those bead lots from their respective data sheets, so that might be nice to include. I've already got scanned digital copies of the data sheets for the following lots: AC03, AE01, AF02, AG01, AH01, AH02).

Thanks!

-John

Jake Beal

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Jan 9, 2017, 7:14:14 AM1/9/17
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Hi, John:

I've been looking up some of this myself, as I've been improving my analytics software to differentiate between different bead lots. The bead lot does play an unfortunately large role in the values observed, up to about 20% variation in the reported values.

I've placed all of the ME* material that I've gathered from the SpheroTech and Clontech data sheets into a new directory in the Google Drive folder.  I've also systematized some of it into an Excel spreadsheet, BeadCatalog.xls, which is formatted to be easily parsed by analytics software.

Speaking of which... does anybody know what laser/filter combination SpheroTech used for their blue channels?  In the email to Brian that's recorded in the Drive, I only see 488nm and 561nm channels, which wouldn't be their BFP or Pacific Blue.

Thanks,
-Jake


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Jake Beal

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Jan 9, 2017, 7:53:26 AM1/9/17
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By the way, an additional complication that I have noticed regarding the multi-fluorophore beads: unlike what one might have reasonably assumed, the relative location of different peaks is often different from channel to channel.

For example, in SpheroTech Lot AA01, the two top brightest beads have the following values in FITC and PE-Cy5 channels:

FITC (488, 530/40):  126914 291016
PE-Cy5 (488, 680/30): 261672 1069885

The FITC/PE-Cy5 ratio for bead 7 is 0.49, while that for bead 8 is 0.27.

While, I do not believe that this is a blockage to using the beads for translation, it again points out that these beads are a complex material, and that we really need to use them *only* based on their actual characterized values and not on projection into behaviors that have not actually been measured.

Thanks,
-Jake

On Mon, Jan 9, 2017 at 6:14 AM, Jake Beal <jake...@gmail.com> wrote:
Hi, John:

I've been looking up some of this myself, as I've been improving my analytics software to differentiate between different bead lots. The bead lot does play an unfortunately large role in the values observed, up to about 20% variation in the reported values.

I've placed all of the ME* material that I've gathered from the SpheroTech and Clontech data sheets into a new directory in the Google Drive folder.  I've also systematized some of it into an Excel spreadsheet, BeadCatalog.xls, which is formatted to be easily parsed by analytics software.

Speaking of which... does anybody know what laser/filter combination SpheroTech used for their blue channels?  In the email to Brian that's recorded in the Drive, I only see 488nm and 561nm channels, which wouldn't be their BFP or Pacific Blue.

Thanks,
-Jake

On Sun, Jan 8, 2017 at 9:06 PM, John Sexton <john.t...@rice.edu> wrote:
Howdy folks. I've been playing around with the mini inter-lab data, and I was unable to find documentation of the bead lots used by David and Ariel in their respective experiments. If you guys could let me know or update your documentations, that would be much appreciated! (sorry if it's there and I missed it!)

(I ultimately need the MEFL and MECY table values for each of those bead lots from their respective data sheets, so that might be nice to include. I've already got scanned digital copies of the data sheets for the following lots: AC03, AE01, AF02, AG01, AH01, AH02).

Thanks!

-John

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Ariel Haim Hecht

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Jan 9, 2017, 11:45:15 AM1/9/17
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My bead lot was AF02.

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Ross, David J. (Fed)

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Jan 9, 2017, 12:40:36 PM1/9/17
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The bead lot for NIST Gaithersburg is AH02.

Jake Beal

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Jan 9, 2017, 12:43:33 PM1/9/17
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That's nice; both are expected to have the same peaks.

What were the bead lots for MIT and Rice?

Thanks,
-Jake

On Mon, Jan 9, 2017 at 11:40 AM, Ross, David J. (Fed) <david...@nist.gov> wrote:

The bead lot for NIST Gaithersburg is AH02.

 

From: sbsc-flow-cytometry@googlegroups.com [mailto:sbsc-flow-cytometry@googlegroups.com] On Behalf Of Ariel Haim Hecht
Sent: Monday, January 09, 2017 11:45 AM
To: sbsc-flow-cytometry@googlegroups.com
Subject: Re: [sbsc-fc] Documenting Bead Lots in Experiment Descriptions

 

My bead lot was AF02.

On Jan 8, 2017, at 7:06 PM, John Sexton <john.t...@rice.edu> wrote:

 

Howdy folks. I've been playing around with the mini inter-lab data, and I was unable to find documentation of the bead lots used by David and Ariel in their respective experiments. If you guys could let me know or update your documentations, that would be much appreciated! (sorry if it's there and I missed it!)

 

(I ultimately need the MEFL and MECY table values for each of those bead lots from their respective data sheets, so that might be nice to include. I've already got scanned digital copies of the data sheets for the following lots: AC03, AE01, AF02, AG01, AH01, AH02).

 

Thanks!

 

-John

 

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Nicholas DeLateur

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Jan 9, 2017, 12:47:37 PM1/9/17
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lot AH01. This information is also contained in my electronic lab notebook on the google drive with the data acquired. 

John Sexton

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Jan 9, 2017, 1:11:19 PM1/9/17
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Sebastian documented the lot number in his FlowCal files. Specifically, for both his "20160918" experiment and his "20161010" the lot number was AH01. You'll see "AH01-06" written in his FlowCal XLSX files because we assign an additional unique identifier per bottle (the "-06" in this case).

Thanks all.

-John

On Mon, Jan 9, 2017 at 11:47 AM Nicholas DeLateur <nichola...@gmail.com> wrote:
lot AH01. This information is also contained in my electronic lab notebook on the google drive with the data acquired. 

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