DropBase:droplet electrosorting 6
Overview
Description
Fibre-FADS device enabling droplet detection based on laser light delivered to the channel by a pair of optical fibres.
Reference
Matthew Penner, Oskar James Klein, Maximillian Gantz1, Friederike E. H. Nintzel, Anne-Cathrin Prowald, Sally Boss, Paul Barker, Paul Dupree, Florian Hollfelder.
Fluorogenic, sub-single-turnover monitoring of enzymatic reactions involving NAD(P)H provides a generalised platform for directed ultrahigh throughput evolution of biocatalysts in microdroplets
unpublished
Submitted by: Matt Penner
Developed by: Tomasz Kamiński & Marta Napiorkowska
Downloads
- Fibre-FADS chip; zip
link temporarily broken due to site maintenance: please find chip design here: https://www.dropbox.com/scl/fi/0cbkkc632ezvvwhw6y5vs/FibreFads.zip?rlkey=epvh2y5o2y2545fh636aakvzj&st=31rzchex&dl=1
Usage Notes
Please enter any comments that other users may find useful below this note (such as flow rates that worked well for particular oil/aqueous phases). When providing usage notes please provide as much detail as useful. We would request that you 'sign' any comments with your initials.
- Using this chip, we found that spacing oil flow rates of around 40 ul/min were a good starting point, along with droplet flow of around 2 ul/min and offset between 1-10 ul/min. The droplet size range tested is between 20-33 pl, but we have found that the chip seems to work better at the higher end of this range. Any higher, and you will get the droplets being split in two by the spacing oil. Any lower, and you will get droplets 'stacking' in the sorting channel. Our work used a laser with wavelength 375 nm and a 380 nm filter. We noted that it is important to excite the droplet using the fibre that is NOT facing towards the incoming droplets. This is to avoid photobleaching, which is actually visible on the trace using this incorrect configuration. We also found (not surprisingly) that the fibres must not have an excessive bend at any point as this may lead to light loss. - MP
