Vhx 900

PHH/HSC sheets were imaged on day 11 using digital microscopy (VHX-900, KEYENCE Ltd., Osaka, Japan) and optical coherence tomography (IVS-2000, KEYENCE Ltd.). The area of the PHH/HSC sheets was analyzed using ImageJ ver. 1.53 (NIH, Bethesda, MD, USA). The sheet thickness of was calculated from the cross-sectional area by randomly imaging three different areas in each sheet using optical coherence tomography.

The appearance of the samples after the Charpy test was taken through a digital microscope, VHX-900 (Keyence corporation, Osaka, Japan), and digital camera. The microstructures were observed through a scanning electron microscope (SEM), VE-7800 (Keyence Corporation, Osaka, Japan), operated at 15 kV. The electron backscattered diffraction analysis was conducted using a 7001F (JEOL, Tokyo, Japan) equipped with a TSL-OIM analytical system.
For each steel bar, full-size 2 mm V-notched specimens with a notch radius of 0.25 mm were machined along the RD, as shown in Figure 3b, and instrumented Charpy impact tests were conducted under a 500 J capacity using CIEM-500D (Tokyo Koki Testing Machine Co., LTD., Sagamihara, Japan). The P and u during the impact test were recorded every 2.0 μs. The value of DBTT was determined from the Charpy curve, i.e., DBTT denoted the absorbed energy transition temperature corresponding to the half value of the vEu.

To fabricate the micro-nanofluidic device, electron beam lithography (ELS-7500; Elionix, Tokyo, Japan) and the dry etching apparatus (NLD-550; ULVAC Co., Ltd., Kanagawa, Japan) were used. A pH meter (F-72; HORIBA, Kyoto, Japan) and an ultrapure water system (Direct-Q UV3; Merck, Tokyo, Japan) were used to prepare the HEPES and PBS buffers. A digital microscope and a fluorescence microscope equipped with a CCD camera (VHX-900 and VB-7010, respectively; KEYENCE, Osaka, Japan) were used to observe the fluorescence. The constant voltage applied to the sorting device came from a high-voltage power supply (HVS448LC-6000D; LabSmith, CA, USA). A UV/O₃ apparatus (Filgen, Aichi, Japan) was used for bonding the silica device and the PDMS reservoir. The particle sizes before and after the proposed sorting were measured using qNano (IZON Science Ltd., Christchurch, New Zealand). A dynamic light scattering (DLS) apparatus (ELSZ-DN2; Otsuka Electronics Co., Ltd., Osaka, Japan) was used to measure the sizes of fluorescent NPs (Figures S2–S5). A tube rotator (TR-350, AS ONE Co., Ltd., Osaka, Japan) and aluminum block bath with a cool-thermo unit (CTU-mini, TAITEC Co., Ltd., Saitama, Japan) were employed for labeling exosomes

DsRNA synthesis and microinjection into larvae were performed as described previously^{19 (link)}. In brief, the cloned cDNA fragments in DNA vectors were amplified by PCR using the primers flanked with the T7 promoter sequence (Supplementary Table 6), and used as templates for dsRNA synthesis. Amplified PCR products were separated by electrophoresis on 1% agarose gels and purified using MagExtractor PCR & Gel Clean up kit (Toyobo). DsRNAs were synthesised using the MEGA script T7 kit (Ambion). Approximately 1.4−2.7 μg and 1.4−2.0 μg of the dsRNAs of Ha-pnr and Cs-pnr were injected into 2-day-old forth (final) instar larvae, respectively. Approximately 2.0−2.7 μg and 1.4–2.7 μg of the EGFP dsRNA were injected into H. axyridis and C. septempunctata larvae as negative controls, respectively. For other genes in the initial small screening, approximately 1 µg of dsRNA was injected into each early last instar larva. Different amount of dsRNA for each gene in this range gave no difference in phenotypic effects. In order to give enough time for the completion of pigmentation, images of adults were captured more than 2 days after eclosion using a digital microscope (VHX-900, Keyence).

Dynamic mechanical analysis (DMA) was carried out on DMA/SDTA861e (Mettler-Toledo, Greifensee, Switzerland) and DMA Q800/RSA3 (TA Instruments, New Castle, DE, USA) in a tension mold. The pre-designed nanocone arrays images of the PVA films surface were obtained by field emission environmental scanning electron microscopy (SEM, Quanta 200 FEG, FEI, Hillsborough, OR, USA), Atomic Force Microscope (AFM, Dimension Icon, Bruker, Karlsruhe, Germany), and optical microscopy (VHX-900, Keyence, Osaka, Japan). The transmission spectra of the SMP films were obtained using a UV–VIS spectrophotometer (Shimadzu UV-3600, Shimadzu, Kyoto, Japan). The reflectance of the SMP films with and without nanocone structures was recorded by a home-built reflectance measurement set up.