Data for: PIEZO1-HaloTag hiPSCs: bridging molecular, cellular and tissue imaging

PIEZO1 channels play a critical role in numerous physiological processes by transducing diverse mechanical stimuli into electrical and chemical signals. Recent studies underscore the importance of endogenous PIEZO1 activity and localization in regulating mechanotransduction. To enable physiologically and clinically relevant human-based studies, we genetically engineered human induced pluripotent stem cells (hiPSCs) to express a HaloTag fused to endogenous PIEZO1. Combined with super-resolution imaging, our chemogenetic approach allows precise visualization of PIEZO1 in various cell types. Further, the PIEZO1-HaloTag hiPSC technology allows non-invasive monitoring of channel activity via Ca2+-sensitive HaloTag ligands, with temporal resolution approaching that of patch clamp electrophysiology. Using lightsheet imaging of hiPSC-derived neural organoids, we also achieve molecular scale PIEZO1 imaging in three-dimensional tissue samples. Our advances offer a novel platform for studying PIEZ..., Micropatterned neural rosettes (MNRs) of 140 µm in diameter grown on 19 mm square Arena A CYTOO chips (Cat. No.10-020-00-18) were imaged on a modified adaptive optical lattice light-sheet microscope (AO-LLSM). Prior to all imaging, the microscope was calibrated to correct for optical aberrations from the system. The Cytoo chip was mounted on a custom designed sample holder and immersed in 40 ml of phenol-free MNR culture medium. The excitation (Thorlabs 0.6 NA, TL20X-MPL) and detection (Zeiss 1.0 NA, 421452-9800-000) objectives were also immersed into the imaging medium. The MNRs were imaged with a multiBessel square lattice light sheet with the NAsq of 0.35 and a 0.3/0.4 NA annular mask. The 488 nm, 560 nm and 642 nm lasers were used to visualize SPY505-DNA (DNA, Spirochrome Cat# CY-SC101), SPY555-actin (actin, Spirochrome Cat# CY-SC202), and Janelia Fluor® 635-HaloTag Ligand (Non-Bapta) or Janelia Fluor® 646-BAPTA-AM- HaloTag ligand (PIEZO1), respectively, with power at the back pupil..., , # Data for \"PIEZO1-HaloTag hiPSCs: Bridging Molecular, Cellular and Tissue Imaging\"

This data is organized for some of the analysis and visualization results shown in the paper \"PIEZO1-HaloTag hiPSCs: Bridging Molecular, Cellular and Tissue Imaging\" ([https://doi.org/10.1101/2023.12.22.573117](https://doi.org/10.1101/2023.12.22.573117)):

## Description of the data and file structure

### Schema of data archive

three_plane_stack_data.zip
├── three_plane_stack_data
│ ├── BaptaWT
│ │ └── 140um
│ │ ├── 2022
│ │ │ ├── 0609
│ │ │ │ └── exp_*
│ │ │ │ ├── actin_mask
│ │ │ │ │ ├── *_lumen_edge.tif
│ │ │ │ │ └── *_outer_edge.tif
│ │ │ │ ├── Actin_with_DNA
│ │ │ │ │ └── *.tif
│ │ │ │ ├── HaloLigand
│ │ │ │ │ └── *.tif
│ │ │ │ ├── results
│ │ │ │ │ ├── *_locs.csv
│ │ │ │ │ └── *_locs_filtered.csv
│ │ │ │ └── settings_info
│ │ │ │ ├── *_JSONsettings.json
...