Figure 2 | Scientific Reports

Figure 2

From: Lipopolysaccharide-induced neuroinflammation disrupts functional connectivity and community structure in primary cortical microtissues

Figure 2

Three-dimensional self-assembled neural cultures express stable GcaMP6s fluorescence and exhibit spontaneous activity by DIV 14. (A) Dissociated P1 primary rat cells are seeded into a custom injection molded agarose well. (B) Cells, which form a “microtissue” (tan) in the first 24 h, were transfected with a commercially available viral vector to encode the GcaMP6s calcium indicator (AAV1-hSyn1-mRuby2-GSG-P2A-GcaMP6s-WPRE-pA). (C) Microtissues display progressive expression of baseline calcium fluorescence from DIV 8—DIV14. (D) Time-lapse videos of calcium transients were collected for 4 min. (E) Python code was used to perform semi-automated segmentation of cell bodies. (F) Segmented regions of interest (ROIs) were overlaid onto the 4-min video for single-cell calcium trace extraction. (G) A raster plot of calcium events detected from calcium traces were extracted from segmented videos using the FluoroSNNAP application40.

Back to article page