Tm4000plus

Scanning electron microscopy was used to map the distribution of crystalline substance in the tissue sections using an energy dispersive X-ray analyzer (SEM-EDX, TM4000Plus, Hitachi High-Technologies, Tokyo, Japan). Briefly, the 4 μm thick FFPE tissue sections were deparaffinized with xylene and washed well with 100% ethanol to eliminate wax and xylene remaining on the sections and dried. Then, well-dried sections were coated at 3 nm thickness with osmium using an osmium coater Neoc (Meiwafosis Co., Ltd., Japan) instrument and subjected to SEM-EDX analysis.

As previously described [6 (link)], the LFA kit’s cellulose pad was coated with a modified NanoSuit^®® solution with Tween-20-based components (Nisshin EM Co., Ltd., Tokyo, Japan), mounted on the wide stage of the specimen holder, and then placed under a desktop scanning electron microscope (TM4000Plus, Hitachi High-Technologies, Tokyo, Japan). Backscattered electron detectors, operating at 10 or 15 kV and 30 Pa, were used to capture the images [6 (link)].

The microstructure of the S₂Ce powder and the cement specimen cured at 37 °C with 100% relative humidity (RH) for 28 d after mixing were observed using SEM (TM4000Plus, Hitachi High-Technologies Corp., Tokyo, Japan); the observation was performed under an acceleration voltage of 15 kV and magnification of 1000.

Paraffin sections (20-µm thick) obtained after plunge freezing and freeze substitution with two needles were deparaffinized in xylene, rehydrated, stained with 1.0% uranyl acetate in 70% methanol for 5 min, washed with distilled water, and stained with Reynolds’ lead citrate solution for 3 min. After washing with distilled water, the specimens were dried at room temperature for 2 h. The microscope slides were placed on the wide stage of the specimen holder using adhesive conductive tape and then placed in an LVSEM (TM4000Plus; Hitachi High-Tech, Tokyo, Japan) operating at 15 kV. Topographic images were reconstructed using Hitachi map 3D software (Hitachi High-Tech).

An optical microscope (Keyence VHX-7000, Keyence, Osaka, Japan) was utilized to capture optical images of the printed traces and substrate. Scanning electron microscopy (SEM) images were obtained using a HITACHI tabletop microscope (TM4000Plus, Hitachi High-Technologies Corp., Tokyo, Japan). Photographs and videos of the sensors in the application demonstration were captured with a smartphone camera. The pressure-sensing tests were performed using a force gauge with an auto stage (IMADA). Resistance measurements were recorded using a digital multimeter (KEITHLEY DMM6500, Tektronix Inc., Cedar Hills, OR, USA) with a 2-wire measurement setup. For the demonstration of human pulse wave and finger tape monitoring, the developed sensors were affixed to the hand of a male volunteer (34 years old). The experiments involving the monitoring of human pulse waves and finger tapping were conducted with the approval of the Yamagata University’s institutional review board (R05-16).

To compare the microscopic features of the different fungal structures (hyphae, spores, and vesicles), fresh cultures of the six strains on SDA GC plates were first examined using optical microscopy. The slides were prepared by gently dabbing the surface of the fungal colony with adhesive tape. The tape was then mounted with one drop of lactophenol cotton blue between the slide and the slipcover. Photographs were taken using a DM 2500 (Leica Camera SARL, Paris, France).
Scanning electron microscopy (SEM) was performed using the TM4000 Plus (Hitachi High-Technologies, Tokyo, Japan) microscope via the 15 KeV lens mode 4 with a backscattered electron detector. A fungal colony sample was cut from the Petri dish and placed on a microscopy slide. A volume of 400 μL of 2.5% glutaraldehyde in a 0.1 M sodium cacodylate buffer was poured over the fungal cut for fixation and stored at 30 °C until completely dry. The standardised fungal structures (hyphae, vesicle, sporangiola, merosporangium, and the number of sporangiospores within the merosporangial sack) were measured using a specific tool for the distance measurement included in the TM4000 Plus microscope. The results were represented in a principal component analysis (PCA) computed using the XLSTAT (Addinsoft, Paris, France) software V.2022.4.1.