Filter testing

Thank you for the interest in my poster at CYTO 2017 in Boston about testing dichroics, the 20 handout copies that I had available were all gone so I’ll take that as a good sign.  I’m working on a writeup with more examples and another test to run, but in the meantime have linked my poster here if you wanted to check it out, along with the email contacts for the vendors of the GoSpectro smartphone spectral analyzer:

Let me know if you have any questions, and I’m hoping to add more content in the near future…

Posted in Uncategorized | Leave a comment

NECyto 2017!

Greetings everyone:

The New England Cytometry Users Group’s annual Fall meeting will be held October 19th, 2017, returning to the Mark and Lisa Schwartz Auditorium of the Ragon Institute at 400 Technology Square in Cambridge MA. We hope you will join us! Please refer to this link for more information about the Ragon Institute:
Advanced registration is preferred, participant rate is still $50.00. On-site registration will be available, so if you have a last-minute sort cancellation and you find yourself free, come on by!

We are very honored to host the following speakers:
Bruce Bagwell MD., PhD. Verity Software
Dave Langenau PhD. Dept of Pathology MGH
Kurt Schalper MD., PhD. Yale Medical School Depts of Pathology and Medicine
J. Phillip McCoy, PhD NIH
Margery Pelletier UMass Lowell, Dept. Biomedical Engineering and Biotechnology
David Wilbur, MD. Dept. of Pathology MGH

There will be parking on-site at the 800 Technology Square Garage, rate to be determined. Take a ticket when you enter the garage and tell the attendant on the way out that you were at the meeting.
Doors open for vendor setup at 7 am and attendees are welcome at 8:00 am for breakfast to be provided in 100/200 Technology Square.
Return here for future updates!
All my best regards and hope to see you in October,
Rich Konz
President; New England Cytometry

Posted in Uncategorized | Leave a comment

NECyto 2016 follow-up

Thanks to all of you who attended the 2016 NECyto meeting this year, we hope you enjoy the t-shirts!  Jennifer Wilshire has shared her Forensic Flow talk and a quiz that has “Crimes of FACSing” in it that we didn’t get to during the meeting.  She welcomes you to use the slides and quiz in your own teaching and edit however you need.

Forensic Flow


Posted in Uncategorized | Leave a comment

Fall Meetings Information

Plenty of educational and networking opportunities in Boston this fall!

Online registration is now open for this year’s NECyto Fall Meeting!  We will have 8 talks, lunch, and a vendor fair, returning to the Ragon Institute after our successful meeting last year.  Please find the speaker list, schedule, and additional information (including the link for online registration) here:

While you are in Boston/Cambridge, why not stick around for the weekend for some additional events?  The NERLSCD Meeting will be held from Oct 13 through Oct 15th
NERLSCD 2016 Registration is open!
If you are a manager or director of a core facility, you should consider attending the 11th annual NERLSCD meeting, to be held Oct 13-15, 2016 , in Boston MA.
The North East Regional Life Sciences Core Directors meeting (often called the NERDS meeting) is an outstanding regional forum for core facility administrators, directors, managers and staff to network with colleagues, to learn about biotechnology advances and their applications, and to discuss the challenges of implementing shared research resources.
Registration link:

Finally, the Biacore Users Group meeting will be held on October 14th at the Ragon Institute.  Learn about applications for SPR (Surface Plasmon Resonance) technologies. More details to follow, contact if you are interested.

We hope to see you in Boston this year!
The NECyto Organizing Committee

Posted in Uncategorized | Leave a comment


facs pain

I experienced #10 the other day, and thought it presented a good opportunity to demonstrate the usefulness of spectraviewers and the importance of knowing your reagents. I’ve compiled a list of spectraviewers on this site, feel free to suggest additional links: (, but even then as I have lamented earlier, you can’t always compare your fluorochromes that are proprietary dyes from different vendors.  Biolegend has some cross-vendor spectra, but their selection is not complete (only a portion of the Live/Dead fixable dyes are shown).  The panel selection program Fluorofinder ( can show spectra for each of the dyes in their multi-vendor database, but don’t yet have a spectraviewer tool to view multiple fluors on the same plot (but that should be coming soon).

So to address the comment about fluors in the same channel, lets use BD Biosciences’s spectraviewer to look at the 2 dyes in question, and we’ll add in the next detector up that is used for BV510 on our system.  The colored bands are the default emission filters used on the system with a long pass in between to split the signal.facspain2

You can see that the 440/40 filter only captures just over half of the peaks of BV421 and Pacific Blue signals and catches the tail of spillover from BV510. This prevents you from having to change the filters around with every change in fluorochrome, but you can see that if you always run BV421, you might want to switch to something like 425/20:


This catches the whole peak of BV421  while avoiding the majority of BV510 spillover.  If you look at the spillover you get from BV510 into each of these filters there is a clear difference (same voltages for both plots):


And when you calculate the compensation matrix, the BV421-%BV510 drops from 30.6 for the 440/40 filter to 0.1% for the 425/20 at the same voltages.  Less spillover means less spread, so comparing compensated BV510 stained beads using either filter and keeping the gate in the same position for both plots shows:


This spread makes it harder to distinguish dim double positives, so if we look at single and double positive events, more double positives don’t resolve themselves from the single positives in the 440/40 setup (2.7% vs 0.7%).


(I ran the populations separately so I could tell how much overlap there was).

With so many reagents to choose from, and different vendors using different names for the same thing, even the experts can lose track, and new tools are always welcomed.  If you know of any programs, platforms, or websites that you find helpful in your flow cytometry research, please post it in the comments and I’ll add it to our links.  Be sure to make use of the existing tools to learn about the properties of your reagents, and talk to your core managers to learn about the setup for your particular systems to get the best results from your experiments.

Posted in Uncategorized | Leave a comment

Flow Ornithology?


(That would be a pair of Buteo jamaicensis, but we have also seen a few Falco peregrinus and various members of the Laridae family…)

Posted in Uncategorized | Leave a comment


Two quick meeting announcements to share for an old colleague, and don’t forget about our NECyto meeting on October 6th!

NERLSCD 2015 Registration is open!
If you are a manager or director of a core facility , you should consider attending the 10th annual NERLSCD 2015 meeting, to be held Oct 14-16, 2015 , in Burlington VT.
The NERLSCD meeting (often called the NERDS meeting) is an outstanding regional forum for core facility administrators, directors, managers and staff to network with colleagues, to learn about biotechnology advances and their applications, and to discuss the challenges of implementing shared research resources.
The early bird deadline for discounted registration and hotel rates is Sept 18th, so register soon!
NERLSCD is a regional chapter of the ABRF.

MetroFlow 2015 Meeting Announcement
Registration is now open for the MetroFlow 2015 meeting!
Join us on October 22, 2015 for another great full-day meeting, to be held at the NY Academy of Medicine (1216  5th Avenue , between 102 and 103rd St.)
We have a content-rich meeting planned, with confirmed speakers including:

  • Bob Balderas (BD Biosciences)
  • Chris Springs (University of Toronto)
  • Jennifer Wilshire (MSKCC)
  • Mark Edinger (Q2 Solutions)
  • Mark Fereshteh (Bristol-Myers Squibb)
  • Pratip Chattopadhyay (Vaccine Research Center, NIAID, NIH)
  • Teresa Hawley (The George Washington University)

Topics presented will include multicolor experimental setup and optimization, fluorescent proteins in flow, compensation and standardization, extracellular vesicle detection, sentinel gating, and high throughput analysis.For more information, please visit the MetroFlow website:
We hope to see you there!

The MetroFlow Steering Committee
Peter Lopez
Research Assistant Professor of Pathology
Director, Core Cytometry Facility, Office of Collaborative Science
NYU School of Medicine
540 First Ave Skirball 2nd floor Administration
New York, NY 10016
212.263.0635 (office)
212.263.5907 (lab)
NYUSOM OCS Core Cytometry Online

Posted in Uncategorized | 1 Comment