Super Res Sunday! And we are doing all the techniques:)

Hari Shroff giving the QI keynote on structural illumination microscopy, adaptive optics and new biosensors. Wow!

Light sheet lab. We image anything from cells to plants to shrimp.
@fedegasparoli.bsky.social @excitedemissions.bsky.social are running the machines!

Light Sheet Saturday is here.
@jencwaters.bsky.social and @talley.codes lecture on how to improve your photon budget by illuminating from the side.

AI for image analysis.
@florianjug.bsky.social @bethcimini.bsky.social @damiandn.bsky.social
are showing off the power of the machines:)

Pizza and image analysis. Because there’s no hackin‘ without snackin‘.
@damiandn.bsky.social @bethcimini.bsky.social

Cake and image analysis. True QI bliss:)
@damiandn.bsky.social @bethcimini.bsky.social @emiglietta.bsky.social teaching 3D segmentation.

Looking for samples for our lightsheet microscope and expansion microscopy lab, @fedegasparoli.bsky.social and @lycasworks.bsky.social go on an expedition during low tide.

Hands on TIRF microscopy and machine learning segmentation has students up all night.

Hard to decide what’s better the beautiful weather or the lectures by @talley.codes and @jencwaters.bsky.social on confocal microscopy.
(it’s obviously the lectures)

We are getting serious about analysis. Today Hunter Elliott and @florianjug.bsky.social introduce machine learning and AI. It’s not magic, it’s computers.

Today’s lectures on live cell imaging and fluorescent proteins by @jencwaters.bsky.social and @talley.codes
Every photon counts!

We tried other brands but nothing works quite like a Guinness for our light sheet microscope.
Future collab?

Today Hunter Elliott is lecturing on image processing. Key take-home before lunch: There is no free lunch:)

This morning‘s lecture is about Koehler illumination.
Pop quiz: What happens when you close the condenser aperture?

Check out the fantastic perspective by @jellevda.bsky.social & @prudentlab.bsky.social on our #pearling work, out in the same @science.org issue!

www.science.org/doi/10.1126/...

The longstanding mystery for how mitochondria segregate and position their DNA has been solved!

Read all about it:
www.science.org/doi/10.1126/...

@jclandoni.bsky.social

Here is Juan and I at the 4th of July parade at MBL physiology

Our paper is now out in Science! Super excited to share our discovery that #mitochondria #pearling is the elusive mechanism driving the regular distribution and inheritance of #mtDNA nucleoids 🧬 [1/6]

Delighted to share our preprint describing COMET, a new approach to drift correction for #SMLM, which performs exceptionally well in practice. Check out the benchmarking results compared to other methods! :) Thanks to @lreinkensmeier.bsky.social for a superb effort.

www.biorxiv.org/cgi/content/...

All set for some expanding

🧪 We are excited to share this novel open access paper on Phasor Mixing Coefficient to analyze colocalization, developed by folks @i2janelia.bsky.social and @aicjanelia.bsky.social. This is also the first technical paper jointly published with our sister imaging center @malacridalab.bsky.social!

www.biorxiv.org/content/10.6...

You rock!! Sorry bout that thanks!

Found the unbroken link:

www.biorxiv.org/content/10.6...

@lycasworks.bsky.social developed quantitative expansion (qExM) to estimate endogenous protein quantities in situ. His ExM protocols offer epitope preservation and accessibility so that standard Western Blot antibodies can be used. Also, check out the cristae, supercomplexes, specialized mitos 🤩

🎸Why this is exciting!!🦖

Quantitative super-resolution often requires genetically encoded tags to get labeling sufficient for quantitative imaging. Here with qExM we use commercial antibodies for endogenous targets.

This opens HUGE opportunities to count in tissues, organisms, or clinical samples!

We then used qExM to measure the abundance of mitochondrial respiratory chain complexes in situ!

We measured the abundances for Complex I, III, and IV.

How they differ in absolute abundances in metabolically distinct subpopulations

And even how T-cell activation changes supercomplex density

We validated qExM first on nuclear pore complexes, attempting to quantify the 8 fold symmetry without using this prior knowledge.

We achieved a mean error of just 9.4% 😍🔬🎸

With these tools in place we moved to tackle protein complex abundances that were unknown....

How does qExM work?

We use cryo-fixation + GMA anchoring to boost antibody labeling efficiency, then apply capture-recapture statistics to dual-labeled samples.

The expansion makes epitopes more accessible while reducing label size relative to targets - perfect for quantitative imaging!

Thank you to our amazing group for this project
@sulianamanley.bsky.social
@kmdouglass.bsky.social
@jclandoni.bsky.social
@tittanoferi.bsky.social
@zimmerli.bsky.social