Bz h4xf

An inverted fluorescence microscope (ECLIPSE, Ti-S, Nikon, Co., Ltd.) with an oil immersion objective (100×, Plan Flour, Nikon) was used. Movies of the sample were taken while moving the viewing field, and the number of clusters having various m values was counted. Approximately 1000 clusters were counted in the total for PS samples. For the titania sample, more than 50 clusters were counted because most of the particles formed large aggregates.
Large aggregates of titania samples were observed using a confocal laser scanning microscope (type C2, Nikon) and an all-in-one microscope (BZ-X800, KEYENCE, Osaka, Japan) equipped with an optical sectioning module (BZ-H4XF, KEYENCE, Osaka, Japan).
The particles’ surface structure and elemental analysis were performed using a transmission electron microscope (type S-4800, Hitachi, Tokyo) and a EDS/EDX detector (ULTIM MAX65, Oxford Instruments, Tokyo, Japan) at the Analysis Center, Nagoya City University.

Live yeast cells expressing Vph1-GFP, Atg32-3HA-3mGFP, or Far8-3×GFP, and mito-DHFR-mCherry or Sec63-mCherry were observed using structured illumination microscopy (SIM). Differential interference contrast (DIC) and fluorescence images were obtained under a Pulse-SIM BZ-X800 microscope (Keyence) equipped with a 100 × objective lens (CFI Apochromat TIRF 100XC Oil, NA: 1.49; Nikon), filter sets for GFP and mCherry (BZ-X filter GFP and BZ-X filter TRITC, respectively; Keyence) and an optical sectioning module (BZ-H4XF; Keyence).

We observed the morphology of SPC-induced contracting cells using the inverted microscope CKX53. Live-cell imaging was performed by first mounting HCASMCs onto glass coverslips, and staining the nuclei with DAPI (Sigma Aldrich, St. Louis, MO, USA) for >30 min after sample preparation, as mentioned in Section 2.1. Thereafter, we stained the plasma membranes with PlasMem Bright Red (Dojindo, Kumamoto, Japan) for 5 min or endosomes with 4 μM of FM4-64 (Sigma Aldrich, St. Louis, MO, USA), which can stain in live cells. The cells were then washed with HEPES buffer, and SPC-induced contraction was induced by 30 μM of nitrobenzoxadiazole (NBD)-SPC for 10 min; that is, SPC with guaranteed fluorescence, with NBD bound to the C-6 position of SPC (Cayman, Ann Arbor, MI, USA). Since SPC is a non-fluorescent compound, we evaluated its intracellular behavior using NBD-SPC, which is commonly used to study the metabolism and transport of sphingolipids [17 (link)]. Cell staining and cross-section images of cells were obtained using an all-in-one fluorescence microscope (BZ-X810, KEYENCE, Osaka, Japan) equipped with an optical sectioning module (BZ-H4XF, KEYENCE, Osaka, Japan).