This file contains the datasets for the Nature Communications article “Thermophoretic Migration of Vesicles Depends on Mean Temperature and Head Group Chemistry” by Emma L. Talbot, Jurij Kotar, Lucia Parolini, Lorenzo Di Michele and Pietro Cicuta. DOI: 10.1038/NCOMMS15351.

The dataset contains raw images relating to the motion of vesicles in thermal gradients. Each compressed folder contains full resolution .tiff images of the fluorescence microscopy data for these vesicles with reference to the figure that data was used in within the main manuscript or supporting information. Each pixel is equal to 0.145 micrometers. Data for fluorescence measurements or zeta-potential measurements (Fig. 5 and Supplementary Fig. 9) are given in an excel spreadsheet. Please refer to the paper for further details.

Images are labelled as follows for figures in the main manuscript: Fig 1c – “State_z_zh=X” (e.g. Initial_z_zh=0.05) where “State” can be either initial or steady state and refers to whether the data was recorded at time t=0 min or after steady state was reached, “X” refers to the height fraction z/zh above the cold plate i.e. z_zh=0.00 would be the cold plate.

Fig 1d and Supplementary Fig 8– “DiPhyPC_TcX_ThY_dT=Z“ (e.g. DiPhyPC_Tc15_Th45_dT=3.0K) where DiPhyPC is the lipid type, “X” is the temperature of the cold plate in ˚C, “Y” is the temperature of the hot plate in ˚C, and “Z” is the temperature difference from that of the cold plate.

Fig 3 - “Lipid_Tbar_X_runY_sliceZ” (e.g. DGDG_Tbar_10_run1_slice1) where “Lipid” is the lipid type (DGDG, SQDG, DiPhyPC, DPPC, DMPC, DOPC, DOPG, or POPG), “X” is the mean temperature between the hot and cold plates e.g. 10 refers to 10˚C, “Y” is the run number (between 1 and 5), “Z” is the slice number where slice 1 is at the cold plate and slice 19 at the hot plate.

Fig 4– Folders contain data for “Lipid_T=XC_frameY” (e.g. DMPC_T=6C_frame1), where “Lipid” is the lipid type or mixture, “X” is the temperature of the sample and “Y” is the frame number. Frames were acquired at 10fps.

Fig 6 and Supplementary Fig 7 – “Lipid_TcX_ThY_t=Zmin_sliceA” (e.g. DOPC_Tc25_Th55_t=0min_slice1) where “Lipid” is the lipid type, “X” is the temperature of the cold plate in ˚C, “Y” is the temperature of the hot plate in ˚C, “Z” is the time in minutes and “A” is the slice number where slice 1 is at the cold plate and slice 19 at the hot plate.

Fig 7 – “Lipid1_Lipid2_Tbar_X_runY_sliceZ” (e.g. DOPC_DOPG_Tbar_15_run1_slice1) where “Lipid1” is the first lipid type and “Lipid2” the second lipid type in vesicles of lipid 1/lipid 2/chol. “X” is the mean temperature between the hot and cold plates e.g. 10 refers to 10˚C, “Y” is the run number (between 1 and 5), “Z” is the slice number where slice 1 is at the cold plate and slice 19 at the hot plate.

Fig 8 – Folders contain data for “Lipid_T=XC_Yfps_frameZ” (e.g. DMPC_T=6C_frame1), where “Lipid” is the lipid type or mixture, “X” is the temperature of the sample, “Y” is the number of frames per second images were acquired at, and “Z” is the frame number.

Supplementary Fig 3 – “Lipid_TcX_ThY_z_zh=Z” (e.g. DOPC_Tc25_Th55_z_zh=0.00) where “Lipid” is the lipid type, “X” is the temperature of the cold plate in ˚C, “Y” is the temperature of the hot plate in ˚C, and “Z” refers to the height fraction z/zh above the cold plate i.e. z_zh=0.00 would be the cold plate.

Supplementary Fig 4 – “Lipid_pH_X_TbarY_runZ_sliceA” (e.g. DOPC_pH_5_5_Tbar30_run1_slice1) where “Lipid” is the lipid type, “X” is the pH e.g. 5_5 is pH 5.5, “Y” is the mean temperature between the hot and cold plates e.g. 10 refers to 10˚C, “Z” is the run number (between 1 and 5), “A” is the slice number where slice 1 is at the cold plate and slice 19 at the hot plate. Alternatively “Lipid_chol_TbarY_runZ_sliceA” for membranes containing chol.

Supplementary Fig 5 – Size distribution data are contained in folders specific to the lipid type with images “Lipid_frameX” where “X” refers to the frame number and “Lipid” the lipid type e.g. DGDG. Images were acquired at 10fps at a temperature of 40 ˚C.

Supplementary Fig 6 – “Lipid_TcX_ThY_z=Zum_frameA” (e.g. DGDG_Tc5_Th15_z=8um_frame1) where “Lipid” is the lipid type, “X” is the temperature of the cold plate in ˚C, “Y” is the temperature of the hot plate in ˚C, “Z” is the distance in µm above the cold plate, and “A” is the frame number. Frames were acquired at 10fps.

This record is pending publication in Nature communications ("Thermophoretic Migration of Vesicles Depends on Mean Temperature and Head Group Chemistry. Experimental dataset."). This record is embargoed until publication and it will be updated.