Bal tec scd 500

The implants were washed for 1 min with PBS (phosphate-buffered saline solution, Dulbeco, Biochrom GmbH, Berlin, Germany) and later with 1 ml of 4% glutaraldehyde. Afterwards, the implants were washed three times for 5 min each time with PBS. The dehydration was carried out by means of an ascending series of 30%, 50%, 70%, 90% and 100% ethanol. The implants were then air-dried until ethanol had completely evaporated. The implants were then attached to SEM sample plates (Agar Scientific Ltd, Stansted, Essex, United Kingdom) and stored overnight in a desiccator (Erich Eydam KG, Kiel, Germany). Gold sputtering with a thickness of 5 nm (BAL-TEC SCD 500, Leica Microsystems GmbH, Wetzlar, Germany) and examination using a scanning electron microscope (Philips XL 30 ESEM, Philips GmbH Market DACH, Hamburg, Germany) has been performed. For each implant, five corresponding area at a magnification of 5000 and five area at a magnification of 8000 were recorded. A semi-quantitative evaluation was performed as described by Acil et al. [12 (link)]. According to that, classification of colonization was classified as: (a) grade 1: no microorganisms detectable; (b) grade 2: sparse microorganisms; (c) grade 3: many microorganisms and conglomerates.

The membranes were washed for 1 min with PBS (phosphate-buffered saline solution, Dulbeco, Biochrom GmbH, Berlin, Germany) and later with 1 mL of 4% glutaraldehyde. Afterward, the membranes were washed three times for 5 min, each time with PBS. The dehydration was carried out by means of an ascending series of 30%, 50%, 70%, 90%, and 100% ethanol. The membranes were then air-dried until the ethanol was completely evaporated. The membranes were then attached to SEM sample plates (Agar Scientific Ltd., Stansted, Essex, UK) and stored overnight in a desiccator (Erich Eydam KG, Kiel, Germany). Gold sputtering at a thickness of 5 nm (BAL-TEC SCD 500, Leica Microsystems GmbH, Wetzlar, Germany) and examination using a scanning electron microscope (Philips XL 30 ESEM, Philips GmbH Market DACH, Hamburg, Germany) was performed. For each membrane, five corresponding areas at a magnification of 5000× and five areas at a magnification of 8000× were recorded.

An environmental scanning electron microscope (GeminiSEM 300, Carl Zeiss, Germany) was applied to evaluate the granule morphology at an accelerating voltage of 10.0 kV. Before observation, every starch sample was spread onto circular metal stubs and sputtered with gold in a sputter coater (BAL‐TEC SCD 500, Leica, Liechtenstein).