An MRC5 cell labelled for microtubules and actin. #FluorescenceFriday

Join the biocontrol seminar tomorrow to hear about the fun things we are doing with controlling cells!

For more details, check out the preprint www.biorxiv.org/content/10.1... - or ask anything here! (+here's a bonus movie that didn't quite make the cut) 🌀

Control was also nicely robust - we demonstrate control of nuclear intensity within the same cell, multiple times!

We could! We showed that we can automatically raise or lower the irradiance over time to maintain specific intensities in the cytosol. We can also use this system to evaluate the efficiency of different controller types.

In order to push outcome-driven microscopy further from the controller side, we also used the same approach with LEXY - the light-inducible nuclear export system (Di Ventura lab). We hoped to control precise concentrations in the nucleus or the cytosol, by placing irradiance under PID control.

Of course, what's the use of making little cell robots if you can't control a few at once with some communication with each other? Here, we added an 'active avoidance' system, giving each cell a searchlight and temporarily reversing migration when a collision is imminent.

If we changed the irradiance over time (in-between loops), we could even slow down and speed up the cells, demonstrating even further control and making use of the dose-responsiveness of optogenetics.

We first tested this with directed cell migration, with the 'outcome' being to guide cells to specific migration paths. Using optogenetic recruitment of TIAM1 (Coppey lab), we could automatically update an area of illumination and precisely guide cells to predefined paths, like little tiny robots! 🤖

First, we had to build the smart microscopy platform. We chose a modular structure, with plug-and-play modules for image analysis, control, and microscope bridging - as required by the user. All this is coordinated by a user-specified 'outcome-driven strategy', with a handy UI for ease-of-use.

In our recent preprint, we address this question - combining smart microscopy with optogenetics to present a platform for 'outcome-driven' microscopy, precisely controlling cells to bring them to the same outcome, despite all the variation. www.biorxiv.org/content/10.1...

Advances in microscopy mean we can now do more than just observe biology—we can control it. But how far can we really push this in mammalian cells with all their (beautiful but annoying) heterogeneity? 🧪🔬(🧵)

Expansion Microscopy is always amazing to me - here is a dividing U2OS cell expanded ~10X and imaged using a 25X objective! 🔬 A total protein stain using maleimide gives further context. #FluorescenceFriday

Sure! 🐌

Klaus Hahn has some classic movies of manually directing migration: tinyurl.com/49ftza3h. The Weiner lab use HL60 (v fast) continually illuminating one side: tinyurl.com/mryvy9m7 . Here is an example of our automatic guidance of HT1080 (slower) to specific paths, look out for a bioRxiv upload soon!🌀

Vimentin DNA-PAINT 🌞 in COS-7 cells #FluorescenceFriday