“Mosquito bites transmit a number of pathogens via salivary droplets deposited during blood-feeding, resulting in potentially fatal diseases. Little is known about the genomic content of these nanodroplets, including the transmission dynamics of live pathogens. Here we introduce Vectorchip, a low-cost, scalable microfluidic platform enabling high-throughput molecular interrogation of individual mosquito bites. We introduce an ultra-thin PDMS membrane which acts as a biting interface to arrays of micro-wells. Freely-behaving mosquitoes deposit saliva droplets by biting into these micro-wells. By modulating membrane thickness, we observe species-dependent differences in mosquito biting capacity, utilizable for selective sample collection. We demonstrate RT-PCR and focus-forming assays on-chip to detect mosquito DNA, Zika virus RNA, as well as quantify infectious Mayaro virus particles transmitted from single mosquito bites. The Vectorchip presents a promising approach for single-bite-resolution laboratory and field characterization of vector-pathogen communities, and could serve as a powerful early warning sentinel for mosquito-borne diseases.”
“a Schematic showing the fabrication process of Vectorchip. Laser patterning was used to obtain through-holes in blocks of PDMS. A thin sacrificial layer of photoresist (Microposit™ S1813 G2, DOW Inc., USA) was spread onto a silicon wafer followed by spin-coating a thin film of PDMS on the wafer surface. The laser-patterned body of the chip was then bonded to the thin film of PDMS using plasma-activated PDMS-PDMS bonding. Membrane-bonded chips were obtained by placing the wafer in an acetone bath, where the sacrificial layer of resist was dissolved. b A PDMS block with laser-generated through-holes (diameter 250 μm). c A PDMS chip bonded to a 1.6 μm thin PDMS membrane on a 4-inch silcon wafer. d A Vectorchip with around 3000 wells (dia: 250 μm) is shown. e Chip with well diameter 1.75 mm and a visible PDMS membrane (thickness 1.6 μm). f Microfluidic channel-integrated chips with a PDMS membrane on top. g These wells can be loaded with feeding solution, reaction reagents, and can store mosquito saliva after biting assays. Wells loaded with feeding media (10% sucrose laced with blue food color) shown in the image. Scale bars represent 1 mm in (b), 5 mm in (e), and 2 mm in (f, g).” Reproduced under Creative Commons Attribution 4.0 International License from Kumar, S., Hol, F.J.H., Pujhari, S. et al. A microfluidic platform for highly parallel bite by bite profiling of mosquito-borne pathogen transmission. Nat Commun 12, 6018 (2021).
Figures and the abstract are reproduced from Kumar, S., Hol, F.J.H., Pujhari, S. et al. A microfluidic platform for highly parallel bite by bite profiling of mosquito-borne pathogen transmission. Nat Commun 12, 6018 (2021). https://doi.org/10.1038/s41467-021-26300-0 under Creative Commons Attribution 4.0 International License
Read the original article: A microfluidic platform for highly parallel bite by bite profiling of mosquito-borne pathogen transmission
Leukocyte differentiation and counting are critical for clinical diagnostics but are hindered by the low…
Screening for microbial proteolytic activity is essential in various biotechnological applications, including bioenergy, food processing,…
Understanding how cellular components, especially chromatin and nuclear condensates, respond to mechanical forces during confined…
In droplet microfluidics, high-throughput screening is critical for analyzing large cellular or molecular libraries at…
In the ever-evolving landscape of biochemical research, protein complexes characterization plays an important role in…
Understanding of a protein’s true behavior in biological systems remains a cornerstone for understanding biological…