Adrian Liston
@labliston.bsky.social
Professor of Pathology at the University of Cambridge. Equalities Fellow, Welfare Tutor and Director-of-Studies at St Catharine's College. Educator, scientist, immunologist, daddy. Founder and CSO of Aila Biotech Ltd.
Fascinating talk tonight from Malala Yousafzai
November 2, 2025 at 9:52 PM
Fascinating talk tonight from Malala Yousafzai
The cumulative effect of these improvements (and others not mentioned!) is enormous. For tough samples, like lung, incorrectly assigned signals are reduced by up to 9000-fold, and a 10- to 3000-fold improvement is common. Look at this ground truth experiment, showing wildtype vs GFP lung:
October 28, 2025 at 8:06 AM
The cumulative effect of these improvements (and others not mentioned!) is enormous. For tough samples, like lung, incorrectly assigned signals are reduced by up to 9000-fold, and a 10- to 3000-fold improvement is common. Look at this ground truth experiment, showing wildtype vs GFP lung:
Not often discussed, but fluorophores also emit differently on different cells. Just as with autofluorescence we can test the different fits on a per cell basis, and use the fit that leaves the least residual. It means more signal is attributed to the correct fluorophore!
October 28, 2025 at 8:06 AM
Not often discussed, but fluorophores also emit differently on different cells. Just as with autofluorescence we can test the different fits on a per cell basis, and use the fit that leaves the least residual. It means more signal is attributed to the correct fluorophore!
We fix this by identifying all autofluorescence patterns in the unstained sample, then applying each pattern to each individual cell in the real sample. We select the autofluorescence index that leaves the least residual, subtract that signal and unmix the rest. The results are gorgeous!
October 28, 2025 at 8:06 AM
We fix this by identifying all autofluorescence patterns in the unstained sample, then applying each pattern to each individual cell in the real sample. We select the autofluorescence index that leaves the least residual, subtract that signal and unmix the rest. The results are gorgeous!
Second, autofluorescence variation. An unmixing matrix that doesn't account for autofluorescence will force all signal into one of the flurophore channels, giving misassigned signal. Past approaches only use autofuorescence index, which means heterogenous mixtures have cells with misassigned signal
October 28, 2025 at 8:06 AM
Second, autofluorescence variation. An unmixing matrix that doesn't account for autofluorescence will force all signal into one of the flurophore channels, giving misassigned signal. Past approaches only use autofuorescence index, which means heterogenous mixtures have cells with misassigned signal
First, pos-neg gating. Ideal controls are your actual antibodies on the actual cells. However different antibodies bind different cells (see: CD4 vs XCR1 below), meaning they have different backgrounds to control. AutoSpectral matches the FSC-SSC of each pos to a suitable neg: with results!
October 28, 2025 at 8:06 AM
First, pos-neg gating. Ideal controls are your actual antibodies on the actual cells. However different antibodies bind different cells (see: CD4 vs XCR1 below), meaning they have different backgrounds to control. AutoSpectral matches the FSC-SSC of each pos to a suitable neg: with results!
Three addition problems:
1)This unmixing solution still requires ideal positive-negative matching to find the right linear regression.
2) Cells have variation in background fluorescence
3) Fluorophores actually stuck on cells have variation in emissions
#AutoSpectral solves these!
1)This unmixing solution still requires ideal positive-negative matching to find the right linear regression.
2) Cells have variation in background fluorescence
3) Fluorophores actually stuck on cells have variation in emissions
#AutoSpectral solves these!
October 28, 2025 at 8:06 AM
Three addition problems:
1)This unmixing solution still requires ideal positive-negative matching to find the right linear regression.
2) Cells have variation in background fluorescence
3) Fluorophores actually stuck on cells have variation in emissions
#AutoSpectral solves these!
1)This unmixing solution still requires ideal positive-negative matching to find the right linear regression.
2) Cells have variation in background fluorescence
3) Fluorophores actually stuck on cells have variation in emissions
#AutoSpectral solves these!
We've all seen the problems - spreading, skewing, autofluorescence intrusion. Unmixing errors are so ubiquitous in high parameter panels they are often thought of as unavoidable, intrinsic to the way the hardware works.
Surprisingly, they are largely artefacts of the unmixing software being used.
Surprisingly, they are largely artefacts of the unmixing software being used.
October 28, 2025 at 8:06 AM
We've all seen the problems - spreading, skewing, autofluorescence intrusion. Unmixing errors are so ubiquitous in high parameter panels they are often thought of as unavoidable, intrinsic to the way the hardware works.
Surprisingly, they are largely artefacts of the unmixing software being used.
Surprisingly, they are largely artefacts of the unmixing software being used.
Pre-print alert! And this one really is a must read for anyone that does spectral #flowcytometry. It is a complete, fully-automated spectral unmixing pipeline that reduces error up to 9000-fold.
Want to get a tough sample, like lung, looking like this? Read more!
www.biorxiv.org/content/10.1...
Want to get a tough sample, like lung, looking like this? Read more!
www.biorxiv.org/content/10.1...
October 28, 2025 at 8:06 AM
Pre-print alert! And this one really is a must read for anyone that does spectral #flowcytometry. It is a complete, fully-automated spectral unmixing pipeline that reduces error up to 9000-fold.
Want to get a tough sample, like lung, looking like this? Read more!
www.biorxiv.org/content/10.1...
Want to get a tough sample, like lung, looking like this? Read more!
www.biorxiv.org/content/10.1...
Another chat to my students about not using AI for research!
October 16, 2025 at 9:30 AM
Another chat to my students about not using AI for research!
Ouch! @xkcd.com hits some truth.
Honestly, it is way too common to see academics pick the “path of ruin”. It is okay for us to not be superstars in our field :)
imgs.xkcd.com/comics/physi...
Honestly, it is way too common to see academics pick the “path of ruin”. It is okay for us to not be superstars in our field :)
imgs.xkcd.com/comics/physi...
October 16, 2025 at 5:59 AM
Ouch! @xkcd.com hits some truth.
Honestly, it is way too common to see academics pick the “path of ruin”. It is okay for us to not be superstars in our field :)
imgs.xkcd.com/comics/physi...
Honestly, it is way too common to see academics pick the “path of ruin”. It is okay for us to not be superstars in our field :)
imgs.xkcd.com/comics/physi...
Completely independent of this, we had the field of regulatory T cells. There were some misleading experiments on "suppressive T cells" early on, but Nicole Le Douarin had used chicken/quail chimeras to prove that some T cells worked to suppress immune responses in the periphery.
October 6, 2025 at 11:21 AM
Completely independent of this, we had the field of regulatory T cells. There were some misleading experiments on "suppressive T cells" early on, but Nicole Le Douarin had used chicken/quail chimeras to prove that some T cells worked to suppress immune responses in the periphery.
A small primer on the #NobelPrize awarded to Mary E. Brunkow, Fred Ramsdell and Shimon Sakaguchi today. This prize was for combining two separate fields of immunology research - genetic research on IPEX and immunology research of regulatory T cells (#Tregs), with enormous impact on biology/medicine
October 6, 2025 at 11:21 AM
A small primer on the #NobelPrize awarded to Mary E. Brunkow, Fred Ramsdell and Shimon Sakaguchi today. This prize was for combining two separate fields of immunology research - genetic research on IPEX and immunology research of regulatory T cells (#Tregs), with enormous impact on biology/medicine
I don't know anyone who worked harder to make the most of opportunities or who learned more set-backs.
James taught me the heart of science. To go head-to-head with the unknown with determination and experience, treating every failure as a challenge to try harder and try smarter.
James taught me the heart of science. To go head-to-head with the unknown with determination and experience, treating every failure as a challenge to try harder and try smarter.
October 1, 2025 at 7:41 AM
I don't know anyone who worked harder to make the most of opportunities or who learned more set-backs.
James taught me the heart of science. To go head-to-head with the unknown with determination and experience, treating every failure as a challenge to try harder and try smarter.
James taught me the heart of science. To go head-to-head with the unknown with determination and experience, treating every failure as a challenge to try harder and try smarter.
Today I'm celebrating Prof James Dooley being made Research Professor at the University of Cambridge!
It is hard to say just how incredible his journey has been. James wrote about his story here, from foster kid to research professor:
issuu.com/adrian.listo...
It is hard to say just how incredible his journey has been. James wrote about his story here, from foster kid to research professor:
issuu.com/adrian.listo...
October 1, 2025 at 7:41 AM
Today I'm celebrating Prof James Dooley being made Research Professor at the University of Cambridge!
It is hard to say just how incredible his journey has been. James wrote about his story here, from foster kid to research professor:
issuu.com/adrian.listo...
It is hard to say just how incredible his journey has been. James wrote about his story here, from foster kid to research professor:
issuu.com/adrian.listo...
Great to see alumni Dr Orian Bricard on the big stage here in Oxford!
September 30, 2025 at 10:36 AM
Great to see alumni Dr Orian Bricard on the big stage here in Oxford!
Oriel College, Oxford
September 29, 2025 at 5:52 PM
Oriel College, Oxford
Misty morning over the Cam
September 29, 2025 at 6:02 AM
Misty morning over the Cam
For the tandem dyes, Tandem Stabilizer is good, but you can make it easier through panel design. Tandem breakdown is not purely chemical - it is higher on monocytes than lymphocytes, and is largely abolished in fixed cells. So move those tandem dyes to post-fix T cells if you can!
September 25, 2025 at 5:06 PM
For the tandem dyes, Tandem Stabilizer is good, but you can make it easier through panel design. Tandem breakdown is not purely chemical - it is higher on monocytes than lymphocytes, and is largely abolished in fixed cells. So move those tandem dyes to post-fix T cells if you can!