Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis - Imaging Flow Citometry Data

Imaging flow citometry (IFC) datasets analysed in "Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis" (under revision).

The folders contain acquisitions of giant unilamellar vesicles (GUVs) for lipid exchange and content exchange controls, with file naming convention DATE_SAMPLE_REPLICATE.rif, content exchange is indicated by CE samples in the 20230228_CE.zip folder, lipid exchange by LE samples in the 20221222_LE.zip folder. 24 samples per set are included, triplicates of isolated P1 (DOPE Af488 0.6% in LE; Dex-Af488 40 uM for CE), P2 (DOPE Cy50.6% in LE; Dex-Af647 10 uM for CE), NC (P1 + P2 1:1), PC (DOPE Af488 0.3% + DOPE Cy5 0.3 in LE; Dex-Af488 20 uM + Dex-Af647 5 uM for CE), and M samples numbered 1 to 4, prepared by mixing P1, P2 and PC in different ratios (M1= 1:1:1; M2= 1:1:0.5; M3= 1:1:0.1; M4= 1:1:0.05).

Only .rif files are provided, they have to be elaborated via compensation and application of an analysis template using the Amnis IDEAS software. Compensation matrices for lipid exchange (20230217_LEcom.ctm) and content exchange (20230217_CEcomp.ctm) are included, as well as the analysis template (Lipid_exchange_analysis_6.2.ast). Gating in the latter may have to be adjusted to analyse LE and CE experiments.

10000 objects in the GUV population or 50000 objects in total were acquired in each file. The files were elaborated in batch mode, outputting the statistic reports (Statistics report CE.txt for CE; Statistics report LE.txt for LE) that were elaborated using an R scirpt (included, IFC_analysis.R)