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Reposted by Juanma García-Arcos

Claire Dessalles @clairedessalles.bsky.social · 1d

All you've ever wanted to know about FLIM analysis (and its application in Flipper-TR)!😱🤩🔬

Juanma García-Arcos @juanmagararc.bsky.social · 3d

🚨 New preprint out!
“Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM” with Tithi Mandal and @rouxlab.bsky.social
We benchmark and describe step-by-step Flipper-TR FLIM measurements across different set-ups and and strategies.

www.biorxiv.org/content/10.1...

Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM

Flipper-TR is a membrane dye sensitive to lipid packing widely used to probe membrane tension in live cells via fluorescence lifetime imaging microscopy (FLIM). However, no consensus currently exists ...

www.biorxiv.org

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Reposted by Juanma García-Arcos

Spirochrome @spirochrome.com · 1d

An incredibly comprehensive and meticulous study of Flipper-TR lifetime analysis to make your membrane tension measurements using Flipper-TR more robust and reliable.

Thanks to @juanmagararc.bsky.social and @rouxlab.bsky.social for this very valuable resource!

Juanma García-Arcos @juanmagararc.bsky.social · 3d

🚨 New preprint out!
“Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM” with Tithi Mandal and @rouxlab.bsky.social
We benchmark and describe step-by-step Flipper-TR FLIM measurements across different set-ups and and strategies.

www.biorxiv.org/content/10.1...

Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM

Flipper-TR is a membrane dye sensitive to lipid packing widely used to probe membrane tension in live cells via fluorescence lifetime imaging microscopy (FLIM). However, no consensus currently exists ...

www.biorxiv.org

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Reposted by Juanma García-Arcos

Joachim Goedhart @joachimgoedhart.bsky.social · 12d

Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM by Juan Manuel Garcia-Arcos and team: www.biorxiv.org/content/10.1...

Frequency dependence of fast-lifetime FLIM and fits for C2C12 cells.

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Reposted by Juanma García-Arcos

KOJAMF🤘🖤🤘 @kojamf.bsky.social · 3d

Dr. Jane Goodall filmed an interview with Netflix in March 2025 that she understood would only be released after her death.

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

*ALL*source data and Python notebooks are available (you can reproduce all figures!)
🔗 doi.org/10.6084/m9.f...
We hope this helps make lifetime analysis transparent and comparable across setups.
📄 www.biorxiv.org/content/10.1...

@rouxlab.bsky.social
#FLIM #FlipperTR #Microscopy #OpenScience

Data and codes

Data and codes corresponding to the preprint:

doi.org

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

Even at realistic photon-limited conditions (~5–10 photons/pixel):
• Phasor analysis remains robust and resilient to noise, although harder to quantify in commercial software.
• Fast FLIM gives clean kinetics but depends on setup
• Fitting yields the most amount of information in ideal situations

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

We also compared Fast FLIM, Fitting and Phasor estimators during osmotic shocks, a direct test of membrane tension.
All track the expected lifetime shifts, but with distinct sensitivities and noise behaviors.

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

We quantified the shot-noise constant (c), the factor translating photon number (N) to lifetime standard deviation (σ): σ ∝ c N⁻⁰·⁵.
📊 Shot-noise constant results: Leica yields consistent photon efficiency; PicoQuant varies by estimator.

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

We show the behavior of tau average and tau 1 across a range of packing standards (GUV with known lipid compositions), showing that tau average has less variability than tau 1.

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

Flipper-TR is now a standard probe for lipid packing and membrane tension, but everyone analyses it differently.
We compared:
• Fast FLIM (mean photon arrival)
• Bi-exponential reconvolution & tail fits
• Phasor analysis
…across photon counts, repetition rates (20–80 MHz), and different set-ups.

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

We first review what we know about the photophysics of the Flipper-TR probe and how this lipid packing probe can be used as a tension reporter

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Juanma García-Arcos @juanmagararc.bsky.social · 3d

🚨 New preprint out!
“Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM” with Tithi Mandal and @rouxlab.bsky.social
We benchmark and describe step-by-step Flipper-TR FLIM measurements across different set-ups and and strategies.

www.biorxiv.org/content/10.1...

Fluorescence lifetime estimation: a practical approach using Flipper-TR FLIM

Flipper-TR is a membrane dye sensitive to lipid packing widely used to probe membrane tension in live cells via fluorescence lifetime imaging microscopy (FLIM). However, no consensus currently exists ...

www.biorxiv.org

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Reposted by Juanma García-Arcos

Spirochrome @spirochrome.com · Feb 6

📢New Actin Probe available at Spirochrome!

Introducing SPY650-FastAct_X our latest actin probe with highly improved kinetics and no toxicity!

spirochrome.com/product/spy6...

The product is now available from our distributors

Preprint from the @kjohnsson.bsky.social lab: doi.org/10.1101/2025...

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Reposted by Juanma García-Arcos

Pablo J. Sáez @pjsaez.bsky.social · Jan 18

How cells decide where to go? #CellMigration

Last year we published our 1st paper @cellcommlab.bsky.social, showing how actin-based membrane oscillations define a new direction ( #cancer & non-cancer #cells )

Full www.nature.com/articles/s41...
@naturephysics.bsky.social @focalplane.bsky.social 🧪🔬

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Reposted by Juanma García-Arcos

Pablo J. Sáez @pjsaez.bsky.social · Feb 6

#CellMigration happens all the time in our bodies. This may (or not) rely on adhesion molecules

#Cancer cells switch strategies during metastasis to invade other tissues

Naomi Wee (naomiwee.com) did this neat movie to explain it:
www.youtube.com/watch?v=D9aD...
#SciSky 🧪🔬 @cellcommlab.bsky.social

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Reposted by Juanma García-Arcos

Ricard Alert Zenón @ricardalert.bsky.social · Jan 15

New preprint! When and how do active flows become turbulent? We found that, in unconfined active nematics, the transition to turbulence is discontinuous! With @the-chaotician.bsky.social @mpipks.bsky.social

arxiv.org/abs/2501.060...

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Reposted by Juanma García-Arcos

Bryan Welm @bryanwelm.bsky.social · Nov 23

I mostly see the protein sculptures I build in the lighting of my workshop. It’s fairly static and dull lighting. When they are close to being finished, I bring them into the house and enjoy watching how they change under different natural lighting throughout the day. They are so much more dynamic!

Light reflecting off a sculpture of different structural domains found in proteins. Made from steel.

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Reposted by Juanma García-Arcos

Itai Yanai @itaiyanai.bsky.social · Nov 22

Would you prefer to lead a lab alone or together with a science buddy? How interesting would be a research institute where joint labs are the standard?!

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Reposted by Juanma García-Arcos

Clausen Lab @clausenlab.bsky.social · Nov 19

i might be biased, but this place is amazing: great colleagues, vibrant campus, brilliant students/postdocs - and Vienna! perfect for starting an ambitious project. apply now for a GL position at the IMP …

www.imp.ac.at/career/open-...

Group leader positions at the IMP | Research Institute of Molecular Pathology

www.imp.ac.at

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Reposted by Juanma García-Arcos

Ute Hellmich @hellmichgroup.bsky.social · Nov 16

Ok, trying to bring my community back together... I created a starter pack for lipids and membrane protein aficionados!

Feel free to share, add and distribute, so we can come together again! 😍

go.bsky.app/AEeXh86

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Reposted by Juanma García-Arcos

Guillaume Jacquemet @guijacquemet.bsky.social · Nov 11

I started a list of people working broadly on cell adhesion and cell migration. I likely missed many, please drop me a message if you want to be added

go.bsky.app/5Gzo7oZ

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Juanma García-Arcos @juanmagararc.bsky.social · Nov 19

Hey super cool
Followed them all
Please add me too !

Reposted by Juanma García-Arcos

MwahahahahahadScientist @mads100tist.bsky.social · Nov 19

I have created a starter pack of scientists in der Schweiz so we can all hopefully find each other 🇨🇭🧪

go.bsky.app/8rF765Z

Scientists in Switzerland starter pack, with our avatars

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Juanma García-Arcos @juanmagararc.bsky.social · Nov 19

Add me please :D

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Reposted by Juanma García-Arcos

Krishna Mudumbi @krishnamudumbi.bsky.social · Nov 17

Sunday #NewPI thoughts

Four panel Star Wars meme of Anakin and Padme in the grass. Anakin looking to his left at Padme. Starts off with Padme smiling but ends with her looking crestfallen

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