Timelapse fluorescence and DIC microscopy of a crawling zoospore

Chytrid fungi make motile spores called zoospores that can crawl like an amoeba or swim with a flagellum. In the Buchler lab, we study a soil chytrid, Spizellomyces punctatus, and use Agrobacterium to insert genes that make the fungi fluoresce or glow (Medina et al, eLife 2020). The video shows a glowing zoospore (left = fluorescence image, right = DIC image), recorded at one frame every 2 seconds. The zoospore expresses a Lifeact-tdTomato fusion protein that highlights the actin filaments at the leading edge of the crawling spore. Scale bar is 5 microns.  If the zoospore were not confined between a glass slide and a gel pad, it could swim at speeds of up to 100 microns per second using the prominent flagellum visible in the DIC image!

References:
Edgar M Medina, Kristyn A Robinson, Kimberly Bellingham-Johnstun, Giuseppe Ianiri, Caroline Laplante, Lillian K Fritz-Laylin, Nicolas E Buchler (2020) Genetic transformation of Spizellomyces punctatus, a resource for studying chytrid biology and evolutionary cell biology eLife 9:e52741