NOCC

Hi and welcome to NOCC project's very own Wiki. Here, Nigar Abbasova (NA) will try to summarise her experimental progress on micropatterning very small fibronectin patterns with topological defects to guide cellular motion.

Enjoy!

Experimental notes

13.06.2024 - NA

19.06.2024 - NA

20.06.2024 - NA

Protocols (to be continued...)

Stenciling protocol (originally by Meng Pan)

Stamping protocol (inspired by Meng Pan, revised by NA)

Short summary for NOCC group meeting 20.06.2024 @UM from NA:

Progress September 2023 - June 2024

Stenciling techniques proves to work best when the intern-pattern distance is anything above 30-50 microns. Anything smaller than that becomes difficult to make a stencil out of.

NA was able to make stencils out of bigger patterns, and even get some data of MDCK motion confined by circular or square patches. After these successful experiments, AAD started preparing wafers with patterns of smaller features for the NOCC project, such as the topological patterns inspired by https://journals.aps.org/pre/abstract/10.1103/PhysRevE.103.022703 amongst other papers.

These patterns have been incredibly difficult to turn into stencils. Some of NA's thoughts on this are:

• it is difficult to make stencils of our micropatterns using our current technique. Using less viscous glue (NOA89 instead of NOA73) did not help much.
• Stamping could be a good alternative - NA is currently testing it out to see if it can work. NA has been doing some stamping with Silja on gels, and has got a bit more practice with stamping protocol. But since we want to stamp our patterns on polymer dishes for now, NA had to modify the protocol a little bit after talking to Meng Pan (MP) about it.

Updates as per 20.06.2024 15:00 Oslo time:

• Stamping that NA tried on 19.06.2024 was not very successful - NA was not able to find any patters on the petri dish... This was the first time executing the protocol, and there are lots of things to be improved.
• However, NA was able to image the PDMS stamps with fluorescent fibronectin to visualise the patterns. The image to the right below shows one of the patterns imaged using fluorescent GPF fibronectin.

Goals for next week (24.06-28.06)

• redo stamping experiments and see if we get better attachment of fibronectin to the petri dishes!

Fluorescent PDMS stamps with GFP fibronectin (prepared on 19.06.2024, imaged on 20.06.2024)

Fluorescent GFP fibronectin on one of the PDMS stamps. From wafer #2 made by AAD. Magnification: 10x

Almost successful stencil of micropattern from AAD's wafer #2. The patterns on the top and bottom of the stamp were transferred well using NOA89 glue, but not the patterns in the middle. This leaves the stencil with a big hole in the middle where there are no patterns, and can therefore not be used further in the experiments.