Full text loading...
Abstract
The use of membrane-specific dyes for in vivo fluorescent microscopy is commonplace. However, most of these reagents are non-specific and cannot track specific lipid species movement, instead often acting as non-covalent lipid-associated probes or requiring the uptake of whole lipids and acyl tails into the membrane. This issue has been solved in eukaryotic cell biology by the use of click-chemistry-liable phospholipid headgroup pulse labels. Here, we describe a method for in vivo phospholipid labelling by fluorescent imaging in Pseudomonas aeruginosa using a phosphatidylcholine mimic, ‘propargyl-choline’ (PCho). This click-chemistry-liable headgroup mimic is visible by microscopy and allows the covalent labelling of lipids. Fluorescence of the cell membranes, visible in heterogeneous patches, is dependent on PCho concentration and is localized in the membrane fraction of cells, demonstrating that it is suitable for membrane labelling and cell imaging.
- Received:
- Accepted:
- Published Online:
Funding
-
Biotechnology and Biological Sciences Research Council
(Award BB/M01116X/1)
- Principle Award Recipient: DavidI Roper
-
Natural Environment Research Council
(Award NE/T014717/1)
- Principle Award Recipient: ChrisLB Graham
-
Antibiotic Research UK
(Award ANTRUK_SRG_05-2021)
- Principle Award Recipient: ChrisLB Graham