Jsm it300 scanning electron microscope

The sample preparation for SEM included hydration in water and dehydration with either 2,2-dimethoxypropane (DMP) [79 (link)] or a series of ethanol (EtOH, 70–85–96%) and critical point dryings. The pollinia/pollinaria of Bulbophyllum retusiusculum, Dendrobium × delicatum, Oncidium crocidipterum, and Polystachya cultriformis were dehydrated according to the DMP Direct Method. The pollinia/pollinaria of Cephalanthera longifolia were dehydrated using a series of ethanol (EtOH, 70–85–96%). All dehydrated samples were transferred into 100% acetone for 10 to 15 min. Critical point drying was performed with an “Autosamdri^®-815—Series A tousimis Critical Point Dryer”. Dried pollinia/pollinaria/anther caps were mounted on aluminium stubs and sputter coated with gold in a “BAL-TEC SCD 050 Sample Sputter Coater”. All samples were investigated using a JEOL JSM-IT300 scanning electron microscope.

Mouse sperm from the cauda epididymis were fixed in 2.5% phosphate-buffered glutaraldehyde (GA) (Zhongjingkeyi Technology, Beijing, China) at room temperature for 30 min and then deposited on coverslips. As described in a previous protocol (Zhang et al., 2022a (link)), the coverslips were dehydrated via an ascending gradient of 50%, 70%, 95%, and 100% ethanol and air-dried. Specimens were then attached to specimen holders and coated with gold particles using an ion sputter coater before being viewed with a JSM-IT300 scanning electron microscope (JEOL, Tokyo, Japan).

Samples for SEM were incubated in 2% glutaraldehyde for 10 min and then stored at −80°C until processing. The samples were then dehydrated with an ethanol dilution series of 30%, 50%, 70%, 80%, 90%, 95% each for 10 min and three times in 100% absolute ethanol for 10 min. The dehydrated samples were CO₂ critical‐point dried with a CPD 300 auto critical‐point dryer (Leica Microsystems). The dried pieces were gold‐coated using a JFC‐2300HR sputter coater (JEOL) for 80 s. Pictures were taken using a secondary electron detector with a JEOL JSM‐IT300 scanning electron microscope with a 15 kV acceleration voltage in ultra‐high vacuum.

The surface morphology of the prepared samples was detected by a JSM-IT300 scanning electron microscope (JEOL Co., Ltd., Tokyo, Japan). Its main technical indicators are (1) Secondary electron image resolution: 3.0 nm. (2) Backscattered electron image resolution: 4.0 nm. (3) Magnification range: 5 times to 300,000 times (continuously adjustable). (4) Accelerating voltage: 0.3~30 kV (multi-level adjustable). The KEYENCE VK-250K laser confocal microscope (KEYENCE Co., Ltd., Osaka, Japan)was used to characterize the three-dimensional topography of the superhydrophobic surface. The laser confocal microscope can obtain the microscopic topography at different depths by fast scanning imaging.
The surface wetting properties were characterized by measuring the static contact angle of water droplets. The surface static contact angle was measured by SDC-80 optical contact angle measuring instrument (Shengding Precision Instrument Co., Ltd., Dongguan, China). The reliability of the measurement data was ensured by measuring the contact angle of the five regions of the sample. The contact angle was obtained by taking pictures of water droplets and analyzing them through software ImageJ-v1.8.0 (National Institutes of Health, Bethesda, MD, USA).

The surface of the lyophilically dried, grafted CC-PBA-PVBE copolymer was studied using a JSM-IT300 scanning electron microscope (SEM) (JEOL Ltd., Tokyo, Japan) with a ≤5 nm diameter electron probe (operating voltage 20 kV), using low-energy secondary electron and back-scattered electron detectors in low vacuum mode to eliminate any charge from the samples.

To study the morphology and the internal structure of CMCh films, a JSM-IT300 Scanning Electron Microscope (JEOL Ltd., Tokyo, Japan) was used. Prior to SEM analysis, the film was fractured in liquid nitrogen and vacuum-sputtered with gold.