Sc7620 mini sputter coater system

At day 14 one scaffold from each condition was removed and divided between SEM and fluorescence imaging. For SEM the scaffold was fixed overnight in glutaraldehyde (2.5% v/v in deionized water) before undergoing sequential dehydration in a graded ethanol series, further dehydration using hexamethyldisilazane and drying overnight. Dehydrated scaffolds were mounted on an aluminum stub using a carbon adhesive disk and gold coated using a SC7620 Mini Sputter Coater System (Quorum Technologies Ltd, East Sussex). High‐resolution images were taken using an environmental scanning electron microscope (ESEM, Carl Zeiss Evo LS15 Variable Pressure Scanning Electron Microscope). Three different areas selected randomly were observed on each sample with three different magnifications (400×, 1000×, and 5000×). Cell coverage at day 14 was calculated using ImageJ by manually drawing around cells and calculating the relative surface area occupied. fiber diameter and pore area were calculated using ImageJ. For fiber diameter 15 fibers per SEM image were measured using ImageJ and the average diameter calculated, and mean fiber diameter for n = 3 scaffolds of each type was calculated. For pore area the image was flattened and binarised before manually drawing around 15 pores per image and calculating average pore area, mean pore area for n = 3 scaffolds of each type was calculated.

The fibres were imaged using SEM as described in our previous study. Samples were mounted on an aluminium stub using a carbon adhesive disk and then gold coated using a SC7620 Mini Sputter Coater System (Quorum Technologies, Ltd, UK). High-resolution images were taken using an environment SEM (Carl Zeiss Evo LS15 Variable Pressure, Germany). Six areas of the fibre mesh were observed and fibre diameters (5 per area, 30 measurements in total) were measured using ImageJ (National Institutes of Health, US) software.

An optical micrometer (Keyence LS-7010MR laser with Keyence LS-7601 monitor, Milton Keynes, United Kingdom) was used to measure suture diameter by averaging five measurements, which was then converted to United States Pharmacopeia (USP) suture sizing standardization. Scanning electron microscopy (SEM) was used to measure fiber diameter. Suture materials measuring 2–3 cm were cut and mounted on aluminum stubs (Agar Scientific, Essex, United Kingdom) using carbon tape (Agar Scientific).
These stubs were coated with gold for 120 s using a SC7620 Mini Sputter Coater System (Quorum Technologies Ltd, Laughton, United Kingdom). SEM images were acquired using an EVO LS15 VP-SEM (Carl Zeiss, EVO LS15, Oberkochen, Germany) in high vacuum mode to examine suture morphology. Fiber diameter was measured from SEM images using ImageJ software (National Institutes of Health, Bethesda, MD). For each experimental condition, 3 samples were imaged and 10 measurements per image were captured at magnifications between 200 × and 1000 × .

Dentine slices and cartilage pieces were fixed in 4% formalin, sputter-coated with gold using the SC7620 Mini Sputter Coater System (Quorum Technologies, Lewes, UK) and imaged using a Philips XL 30/SEM with field emission gun.

Samples were analysed, after gold coating using a SC7620 Mini Sputter Coater System (Quorum Technologies Ltd, East Sussex, UK), by scanning electron microscopy (SEM) using a Zeiss Evo LS15 Variable Pressure Scanning Electron Microscope (Carl Zeiss Microscopy GmbH, Oberkochen, Germany). Images were taken in triplicates at 5000X magnification to allow a minimum of 30 fibres to be measured for their diameter.