Liffner and his team optimized an expansion microscopy workflow to physically enlarge whole mosquitoes, enabling detailed visualization of malaria parasites within their tissues. Left is the mosquito at natural scale; right shows the expanded view composed of hundreds of tiled images, with protein-rich areas (white) and DNA (light blue) highlighting parasite location.
Image credit:Joe van den Bergh, Benjamin Liffner, Adelaide Microscopy.
When Benjamin Liffner, a parasite biologist at Adelaide University, adopted expansion microscopy during his postdoctoral work, he recognized its potential to transform parasite research. “Malaria parasites are barely 1.5 microns wide,” he explains, “and their complex organelles demand advanced imaging techniques.” This realization shifted his focus toward combining parasitology with microscopy expertise.
Liffner’s lab now prioritizes microscopy-driven studies of parasite biology. Their recent breakthrough involved adapting expansion microscopy to whole mosquitoes, preserving anatomical context while detecting *Plasmodium* species inside the insect’s body.
The development required extensive refinement. Initial 2023 protocols imaged parasites in cultured cells, followed by 2024 methods targeting dissected organs. However, whole-mosquito imaging presented unique challenges. “Extracting organs altered the parasite’s natural environment,” Liffner notes. Their solution? Crosslinking the mosquito exoskeleton with hydrogel, allowing controlled hydration to expand the sample for microscopy. After iterated experiments, including a water-immersion imaging setup suggested by Cambridge’s Miguel Cardoso Mestre, the team achieved deep-tissue visualization.
The process involved trial-and-error to balance reagent concentrations and incubation times. Staining for proteins and DNA then revealed parasite locations within the mosquito’s organs. While Liffner initially pursued this as a technical challenge, the results sparked scientific interest. “Seeing parasites in their natural habitat was transformative,” he admits, “and now we hope others use this tool to address unforeseen questions about parasite-host interactions.”
