Jsm 6510

Five individuals of each species from each sampling date were examined they were first dissected in order to acquire spines from the middle section of the interambulacral zones and interambulacral plates near the peristome. The plates and spines were then cleaned of internal organs using a 1% bleach treatment in distilled water for 30 min and dried at 60 °C for 3 days [33 (link)]. Specimens were coated in carbon by a Q150R Plus-rotary pumped coater carbon thread evaporator, reducing picture clarity in comparison to a gold coating, but enabling Energy-Dispersive X-Ray Spectroskopy (Figure 2). The samples were examined and imaged by SEM JEOL JSM 6510. For the cross-section of the spines, the middle part of the shaft was selected. Five observations for each species were carried out to determine the number of wedges. To study the chemical composition of the spines and tests, an EDS analysis was carried out by a JEOL JSM 6510 scanning electron microscope, equipped with an Oxford Link ISIS 300 system. Each measurement lasted 240 sec and was made in a 0.10² mm area. The characterization of the various stereom was carried out according to Smith, 1980.

The ultrastructure of the cECM hydrogel scaffolds was visualized with a JSM-6510 (JEOL, Tokyo, Japan) scanning electron microscope (SEM). cECM scaffolds of 20 × 9 × 3 mm³ were produced via 3D bioprinting casting
The 3D scaffolds were fixed in 4% paraformaldehyde (PFA) in PBS for 48 h and then washed three times with 0.1 M phosphate buffer (PB). Next, the samples were incubated in 4% osmium tetroxide for 90 min and then rinsed with deionized water. Subsequently, samples were dehydrated by washing them with ethanol 80% (×2), 90% (×3), 96% (×3), and 100% (×3) and preserved in absolute ethanol at 4 °C until critical point drying (Autosamdri-815 critical point dryer, Tousimis, Rockville, MD, USA). Samples were then carbon coated and mounted using conductive adhesive tabs (TED PELLA, Redding, CA, USA). Imaging was performed by using an SEM (JSM-6510, JEOL, Tokyo, Japan) at 15 kV.
The diameter of the fibers was assessed following the method developed in [47 (link)]. Briefly, 10 fibers of three different zones of each sample were randomly selected, and their diameter was computed with ImageJ Software v1.53 (National Institute of Health, Bethesda, MD, USA).

To observe the plastic surface erosion or decomposition after microbial degradation, we used scanning electron microscopy [39 (link)] (Jeol, JSM-6510, Tokyo, Japan). The SEM sample preparation protocol followed that of the Joint Center for Instruments and Researchers, College of Bio-Resources and Agriculture, NTU. Plastic films cultured with A. fumigatus L30 or A. terreus HC for 30 days were removed from the flask, and each film was cut into 2 pieces and divided into washed and unwashed groups. For the washed group, the plastic film was washed with distilled water to remove the mycelia attached to the plastic surface. The plastic films were immediately soaked in 2.5% (w/v) glutaraldehyde (Sigma–Aldrich Co.) at 4 °C and then shaken overnight for cell fixation. The fixed films were washed with 0.1 M sodium phosphate buffer (pH = 7.3) and postfixed in 1% osmium tetroxide (w/v) in an ice bath for 1 h. The samples were dehydrated gradually with different concentrations of ethanol (30%, 50%, 70%, 85%, 90%, 95%, 100%) for 60 min at each concentration, except for 70% and 100% ethanol for which overnight dehydration was conducted. The dehydrated samples were placed in a critical point dryer for critical spot drying, and subsequently, the samples were coated with gold and observed under a scanning electron microscope (Jeol, JSM-6510) [40 (link)].