Figure 5: Multi-colour X-RASP in vitro. | Nature Communications

Figure 5: Multi-colour X-RASP in vitro.

From: Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation

Figure 5

(a–c) Diagram depicting the mutations introduced in the spXFP1–10 fragment to obtain yellow and cerulean X-RASP when reconstituting fluorescence with the invariant spXFP11 (represented in pink). (d) In vitro X-RASP fluorescence in HEK293 cells. Each coloured X-RASP was produced by transfecting the syb:spGFP1–10, syb:spYFP1–10 or syb:spCFP1–10 with spXFP11 into separate wells. After 24 h of incubation to allow expression, the cells were mixed together, fixed and imaged by confocal microscopy. Left panels show magnified individual G-, Y- and C-RASP cells. Scale bar, 20 μm. (e) Normalized emission spectra measured by confocal microscopy using spectral detection settings (see Methods for details). C-RASP (blue trace), G-RASP (green trace) and Y-RASP (yellow trace) were obtained by co-transfection of HEK293 cells as in d.

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