fspasqualini.bsky.social
@fspasqualini.bsky.social
Everything is open-science here, links in the pre-print, but reach out if you don't want to wait for the final repositories to be updated. www.biorxiv.org/content/10.1...
CALIPERS: Cell cycle-aware live imaging for phenotyping experiments and regeneration studies
Cell cycle progression, migration, and proliferation shape development and regeneration, but simultaneous live-cell imaging remains challenging as conventional fluorescent cell cycle indicators (FUCCI) monopolize the green and red channels used by most structural and functional biosensors. To overcome this, we integrated a spectrally re-engineered FUCCI variant, open-source analysis software, and four-color human stem cell reporter lines into CALIPERS: a method for Cell-cycle-Aware Live-cell Imaging in Phenotyping and Regeneration Studies. ### Competing Interest Statement The authors have declared no competing interest. European Research Council, https://ror.org/0472cxd90, 852560 European Union, https://ror.org/019w4f821, 101140192 Italian Ministry of Enterprises and Made in Italy, F13C23003260003
www.biorxiv.org
July 8, 2025 at 9:36 AM
Everything is open-science here, links in the pre-print, but reach out if you don't want to wait for the final repositories to be updated. www.biorxiv.org/content/10.1...
By tracking rare cycling hiPSC-derived cardiomyocytes, we can monitor cell proliferation (useful in regeneration) and multinucleation and endoreplication (useful for assessing maturation) as well as structure and function integration #CCAwarePhenotyping #CCAwareRegenerativeStudies #MethodInACellLine
July 8, 2025 at 9:36 AM
By tracking rare cycling hiPSC-derived cardiomyocytes, we can monitor cell proliferation (useful in regeneration) and multinucleation and endoreplication (useful for assessing maturation) as well as structure and function integration #CCAwarePhenotyping #CCAwareRegenerativeStudies #MethodInACellLine
New light-sheet capture of 4-color hiPSC-cardioids reveals compaction before calcium transients initiation. 90 % exit cycle by d4.5 → maturation on cue. #CCAwarePhenotyping #MethodInACellLine
July 8, 2025 at 9:36 AM
New light-sheet capture of 4-color hiPSC-cardioids reveals compaction before calcium transients initiation. 90 % exit cycle by d4.5 → maturation on cue. #CCAwarePhenotyping #MethodInACellLine
Safe-harbor genome editing ensures expression across differentiated lineages, and WTC-11 genetic background means full compatibility with the
@alleninstitute.org Cell Collection: the largest repository of GFP/RFP-based hiPSC reporter lines www.allencell.org/cell-catalog...
@alleninstitute.org Cell Collection: the largest repository of GFP/RFP-based hiPSC reporter lines www.allencell.org/cell-catalog...
July 8, 2025 at 9:36 AM
Safe-harbor genome editing ensures expression across differentiated lineages, and WTC-11 genetic background means full compatibility with the
@alleninstitute.org Cell Collection: the largest repository of GFP/RFP-based hiPSC reporter lines www.allencell.org/cell-catalog...
@alleninstitute.org Cell Collection: the largest repository of GFP/RFP-based hiPSC reporter lines www.allencell.org/cell-catalog...
WTC11-based hiPSCs reporter lines. We provide a 4-color CALIPERS hiPSC line with sensors for actin, calcium, and the cell cycle, as well as a blank canvas line to add your favorite sensors. CALIPERS is a #MethodInACellLine!
July 8, 2025 at 9:36 AM
WTC11-based hiPSCs reporter lines. We provide a 4-color CALIPERS hiPSC line with sensors for actin, calcium, and the cell cycle, as well as a blank canvas line to add your favorite sensors. CALIPERS is a #MethodInACellLine!
Phase-locked motility
Each nucleus gets a %-cell-cycle score that rides with its mask and trajectory. Compare shape or speed by cycle progress, not frame time. Watch HaCaTs sprint through physical space and cell cycle (CC) phases! #PhaseLockedMotility #CCAwarePhenotyping
Each nucleus gets a %-cell-cycle score that rides with its mask and trajectory. Compare shape or speed by cycle progress, not frame time. Watch HaCaTs sprint through physical space and cell cycle (CC) phases! #PhaseLockedMotility #CCAwarePhenotyping
July 8, 2025 at 9:36 AM
Phase-locked motility
Each nucleus gets a %-cell-cycle score that rides with its mask and trajectory. Compare shape or speed by cycle progress, not frame time. Watch HaCaTs sprint through physical space and cell cycle (CC) phases! #PhaseLockedMotility #CCAwarePhenotyping
Each nucleus gets a %-cell-cycle score that rides with its mask and trajectory. Compare shape or speed by cycle progress, not frame time. Watch HaCaTs sprint through physical space and cell cycle (CC) phases! #PhaseLockedMotility #CCAwarePhenotyping
Proposed solution: To overcome this, we integrated a spectrally re-engineered FUCCI variant, open-source analysis software, and four-color human stem cell reporter lines into CALIPERS: a method for Cell-cycle-Aware Live-cell Imaging in Phenotyping and Regeneration Studies.
July 8, 2025 at 9:36 AM
Proposed solution: To overcome this, we integrated a spectrally re-engineered FUCCI variant, open-source analysis software, and four-color human stem cell reporter lines into CALIPERS: a method for Cell-cycle-Aware Live-cell Imaging in Phenotyping and Regeneration Studies.
Problem: Cell cycle progression, migration, and proliferation shape development and regeneration, but simultaneous live-cell imaging remains challenging as conventional fluorescent cell cycle indicators (FUCCI) monopolize the green and red channels used by most structural and functional biosensors.
July 8, 2025 at 9:36 AM
Problem: Cell cycle progression, migration, and proliferation shape development and regeneration, but simultaneous live-cell imaging remains challenging as conventional fluorescent cell cycle indicators (FUCCI) monopolize the green and red channels used by most structural and functional biosensors.
Reposted by fspasqualini.bsky.social
The preprint is now available! Check it out... and... if you want to repost, why not 👇the post down here👇, so others get the bioRxiv link right away? Oh, and... 𝐭𝐡𝐚𝐧𝐤𝐬! 🙏
doi.org/10.1101/2025...
bsky.app/profile/flor...
doi.org/10.1101/2025...
bsky.app/profile/flor...
🔬🎤 As pre-announced a few days ago... please let us proudly present to you: 𝑴𝒊𝒄𝒓𝒐𝕊𝒑𝒍𝒊𝒕 - your ticket to imaging more, imaging more gentle, and/or imaging more efficient. 🔬
doi.org/10.1101/2025...
Like ❤️, repost 🔂, and most importantly... please send feedback ✉️ our way! 🙏
doi.org/10.1101/2025...
Like ❤️, repost 🔂, and most importantly... please send feedback ✉️ our way! 🙏
February 11, 2025 at 6:13 PM
The preprint is now available! Check it out... and... if you want to repost, why not 👇the post down here👇, so others get the bioRxiv link right away? Oh, and... 𝐭𝐡𝐚𝐧𝐤𝐬! 🙏
doi.org/10.1101/2025...
bsky.app/profile/flor...
doi.org/10.1101/2025...
bsky.app/profile/flor...
You are too kind! And we definitely couldn't have done this without the help of your lab and @florianjug.bsky.social's!
February 10, 2025 at 8:46 AM
You are too kind! And we definitely couldn't have done this without the help of your lab and @florianjug.bsky.social's!
Reposted by fspasqualini.bsky.social
HYDRA supports fluorescence microscopy, enabling cell-cycle and cytoskeletal analyses with FUCCIplex sensors using CALIPERS (biorxiv.org/content/10.1...). From cell proliferation to mitotic spindle dynamics, this method works with cutting-edge imaging and image-analysis modalities.
January 17, 2025 at 4:14 PM
HYDRA supports fluorescence microscopy, enabling cell-cycle and cytoskeletal analyses with FUCCIplex sensors using CALIPERS (biorxiv.org/content/10.1...). From cell proliferation to mitotic spindle dynamics, this method works with cutting-edge imaging and image-analysis modalities.