The storage medium of the filamentous bacterial fraction was replaced by Dulbecco's Phosphate Buffered Saline liquid (DPBS, Thermo Fisher Scientific) with 1.5% ultra-low gelling temperature agarose A5030 (Sigma-Aldrich). To reduce the risk of contamination introduced by manual picking of single filaments, bacteria were encapsulated into microdroplets at a concentration of 0.1 particle/droplet by a previously fabricated microfluidic device (19 (link)). The cell suspension with 1.5 × 103 particles/μl and 2% Pico-Surf 1 in Novec 7500 (Dolomite) as carrier oil were loaded to the microchannel through polytetrafluoroethylene (PTFE) tubing (AWG24) by a Mitos P-pump (Dolomite), and droplets (50 µm in diameter) were generated. Collected droplets were solidified on ice and moved into the water phase from the oil phase as previously reported (48 (link)). Trapping the filaments in the gel microdroplets can reduce the risk of contamination caused by the simultaneous aspiration of multiple filaments. Droplets containing single filaments were manually picked using a micropipette (Drumond) under microscopic observation and isolated into a 384-well glass bottom (Corning) plate with 1.5 µl of DPBS.
Dulbecco s phosphate buffered saline liquid dpbs
Manufactured by Thermo Fisher Scientific
Dulbecco's Phosphate Buffered Saline (DPBS) is a balanced salt solution commonly used in cell culture and other laboratory applications. It is a sterile, isotonic buffer that helps to maintain the pH, osmolarity, and ionic composition of the sample or environment. DPBS is available in liquid form.
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Lab products found in correlation
2 protocols using dulbecco s phosphate buffered saline liquid dpbs
1
Bacterial Filament Encapsulation and Isolation
2
Filamentous Bacteria Encapsulation and Isolation
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The storage medium of the filamentous bacterial fraction was replaced by Dulbecco's Phosphate Buffered Saline liquid (DPBS, Thermo Fisher Scientific) with 1.5% ultra-low gelling temperature agarose A5030 (Sigma-Aldrich). To reduce the risk of contamination introduced by manual picking of single filaments, bacteria were encapsulated into microdroplets at a concentration of 0.1 particle/droplet by a previously fabricated microfluidic device 19 . The cell suspension with 1.5 × 10 3 particles/μL and 2% Pico-Surf™ 1 in Novec™ 7500 (Dolomite) as carrier oil were loaded to the microchannel through polytetrafluoroethylene (PTFE) tubing (AWG24) by a Mitos P-pump (Dolomite), and droplets (50 µm in diameter) were generated. Collected droplets were solidified on ice and moved into the water phase from the oil phase as previously reported 43 . Droplets containing single filaments were manually picked using a micropipette (Drumond) under microscopic observation and isolated into a 384-well glass bottom (Corning) plate with 1.5 µl of DPBS.
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