7/7
If you are using mVenus or mCitrine YFPs, consider switching to mGold2s and mGold2t for your imaging experiments!

🧬 Plasmids available on @addgene.bsky.social
🛠️ Need help subcloning for your system? Feel free to reach out
💡 We also have preliminary circular permutation points worth trying!

6/7
➡️ In all cases, replacing photolabile YFPs with mGold2s/t enabled longer imaging without compromising assay performance.

I thank all the co-authors and the funding sources for making this project possible!

5/7

We teamed up with 5 labs to test mGold2s and mGold2t across diverse imaging platforms:

🧠 Imaging actin dynamics (Chen Lab)
🧠 Imaging inhibitory synapses (Tolias Lab)
🧠 cAMP FRET biosensors (Zhang Lab)
🧠 Single-molecule imaging (Ha Lab)
🧠 Widefield microscopy (Shaner Lab)

4/7
🎉 The result? Meet mGold2s and mGold2t:
🟡 ~20–25× more photostable than mVenus and mCitrine
🟡 4× more photostable than our previous champion, mGold
🟡 Retain high brightness and other properties

3/7
To fix this, we used SPOTlight—our pooled, single-cell, microscopy-based screening platform—to screen 1.1 million cells encoding >200,000 variants.

By simultaneously screening for brightness and photostability, we beat the typical trade-off between the two.

2/7
Why do we need more photostable FPs?

Many FPs photobleach rapidly under repeated or prolonged illumination, limiting imaging time and resolution.

Notably, yellow FPs, despite their popularity, photobleach faster than FPs in other spectral classes.

🧵 1/6
Just dropped in @naturecomms.bsky.social

We’ve engineered the most photostable yellow fluorescent proteins (YFPs) to date—mGold2s and mGold2t—with up to 25× greater photostability than mVenus and mCitrine, without compromising brightness.

👇
🔗 www.nature.com/articles/s41...