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Are you looking for a postdoc exploring the cell biology of development, defining how cells change shape and move? Do you want to develop a research program & teaching skills preparing you for a position at a college or small University? Our new NIH grant will support those goals 1/n Please share

Lastly, we were also able to visualize the development of transcriptional asymmetry in a growing microcolony. Using a dual auxin- and cytokinin-signaling reporter-gene construct, it was seen that the reporter expression starts to diverge into distinct domains in more mature microcolonies. (10/n)

We managed to capture the donut-shaped structure of microtubules surrounding a phragmoplast forming between two recently separated nuclei. Thanks to Emry Ng for the image processing! (9/n)

We also used a Lattice Lightsheet microscope to perform high-temporal resolution live imaging of cultured cells. This set up allowed us to catch a dividing cell in the act. Our favorite is an example showing the microtubule dynamics during the first division of a regenerating protoplast. (8/n)

We also showcased the use of additional cellular markers, for the Golgi and microtubules. Confocal Z-stacks of developing microcolonies demonstrate the intricate details of intracellular organization of cellular compartments and components during the early stages of regenerative development. (7/n)

Furthermore, the inclusion of fiducial fluorescent beads embedded within the alginate discs allowed us to find and track the same cells over time. (6/n)

To visualize the development of protoplast regeneration microscopically with enhanced detail and resolution, we incorporate fluorescent cellular markers for the plasma membrane and the nucleus. To better distinguish cell-division events, which can be challenging with brightfield microscopy. (5/n)

Part of the novelty of this method is the embedding of protoplasts in alginate discs that fit into the wells of a 24-well plate. This format “enables high throughput experimental design: a single isolation yields more than 24 discs, each potentially representing an independent treatment.” (4/n)

This work was initiated over seven years ago, when Kelsey first started to develop a protoplast regeneration system using Arabidopsis. There were several methods already published, but we struggled to make them work in our hands. We hope that the method used here will be more transferrable. (3/n)

🌱We are looking for an Assistant Professor in Plant Molecular Genetics @irbv.bsky.social If you are passionate about genetics, molecular biology, or imaging, this is your chance to join a community of researchers working on plant development, cell biology, evolution, ecology, and biodiversity. 👇