Pelco sc 6 sputter coater

Specimens for SEM analyses were preserved in 70% ethanol for several days. For preparation a method modified from that by Kanturski et al. [13 (link)] was used. From ethanol the specimens were transferred into 6% phosphotungstic acid (PTA) solution in 70% ethanol for 24 hours. Dehydration was provided by ethanol series of 80%, 90%, 96% and two changes of absolute ethanol for 30 minutes each. Dehydrated specimens were dried using hexamethyldisilazane (HMDS) solution with absolute ethanol in proportions of 1:3, 1:2; 2:3 for 30 minutes each followed by two changes of undiluted HMDS. Samples were mounted on aluminium stubs with double-sided adhesive carbon tape and sputter-coated in a Pelco SC-6 sputter coater (Ted Pella Inc., Redding, CA, USA). The specimens were imaged by the Hitachi SU8010 field emission scanning electron microscope FESEM (Hitachi High-Technologies Corporation, Tokyo, Japan) at 5, 10 and 15 kV accelerating voltage with a secondary electron detector (ESD). Sensilla terminology follows Bromley et al. [14 , 15 ].

To confirm the presence of the attached cells on the carrier surface during naproxen degradation, scanning electron microscopy (SEM) was used. The carrier with immobilized bacteria was prepared for imaging with SEM using 3% v/v glutaraldehyde incubation (fixative, 24 h) and subsequent ethanol dehydration (30, 50, 70, 80, 90, 95, and 100% v/v, each for 15 min). The samples were subsequently critical-point-dried in the Leica EM CPD300 Automated Critical Point Dryer (Leica Microsystems, Vienna, Austria), mounted on aluminum stubs with double-sided adhesive carbon-tape and sputter-coated in a Pelco SC-6 Sputter Coater (Ted Pella Inc., Redding, CA, USA) with a thin film of gold to improve the electrical conductivity of the sample surface. After processing, samples were imaged using the Hitachi SU8010 field emission scanning electron microscope (FESEM) (Hitachi High-Technologies Corporation, Tokyo, Japan) at 10 kV accelerating voltage with a secondary electron detector (SED) and at a working distance (WD) of 3–300 μm.

Imaging of the bacterial film present on the surface of the nylon monofilaments used in this study was performed with Hitachi UHR FE-SEM SU 8010 scanning electron microscope with an acceleration of 10 kV using a secondary electron detector (SED). Samples of a sterile nylon fibers and fibers immersed in 85 × 10⁶ CFU/ml solution of B. subtilis were placed on aluminum stubs with adhesive carbon tape and allowed to dry completely at 22 °C for 12 h. After this time, the prepared material was covered with a 0.4 mm layer of gold in a Pelco SC-6 sputter coater (Ted Pella Inc., Redding, CA, USA) and observed in a scanning electron microscope.

To confirm the presence of the attached cells on the carrier surface, scanning electron microscopy (SEM) was used. Fragments of the carrier were prepared for imaging with SEM using 3% v/v glutaraldehyde incubation (fixative, 24 h) and subsequent ethanol dehydration (30, 50, 70, 80, 90, 95 and 100% v/v, each for 15 min). The samples were subsequently critical-point dried in the Pelco CPD2 apparatus (Ted Pella Inc., Redding, CA, USA), mounted on aluminium stubs with double-sided adhesive carbon-tape and sputter-coated in a Pelco SC-6 Sputter Coater (Ted Pella Inc., Redding, CA, USA) with a thin film of gold to improve the electrical conductivity of the sample surface. After processing, samples were imaged using the Hitachi SU8010 field emission scanning electron microscope (FESEM) (Hitachi High-Technologies Corporation, Tegama, Japan) at 5, 10 and 15 kV accelerating voltage with a secondary electron detector (SED) and at a working distance (WD) of 3–300 μm.

Apical parts of roots (in R roots, a fragment protruding from the tube) were cut off and immediately fixed in 100% methanol for 1 h at room temperature [28 (link)]. Next, the samples were dehydrated in 100% ethanol for 30 min, transferred to fresh 100% ethanol and left overnight. After dehydration, a critical point drying procedure with the EM CPD300 Automated Critical Point Dryer (Leica Microsystems, Wetzlar, Germany) was done following the ‘Rice Root’ protocol according to the manufacturer’s instructions. Dried apices were mounted on aluminum stubs with double sided adhesive carbon tabs and sputter coated with a 10–15 nm thick gold film in a Pelco SC-6 sputter coater (Ted Pella, Inc., Redding, CA, USA). The specimens were observed using a UHR FE-SEM Hitachi SU8010 field emission scanning electron microscope (Hitachi High-Technologies Corporation, Tokyo, Japan) at accelerating voltages of 7 kV and 10 kV and a working distance of 8–10 mm. In C and S roots, micrographs were collected in the meristematic zone (about 0.5–1 mm from the root tip). In case of R roots, two regions were imaged: below and above the boundary between two shades of Evans Blue stain. Five to seven apices from each of the three groups (C, S, R) were analyzed.