Nova nano 450 sem

Observation of the surface morphology of the Gynemesh and Restorelle was performed using a scanning electron microscope (FEI Nova Nano 450 SEM). Samples were not subject to deposition of conductive coatings, in contrast to usual polymers SEM analysis practice. To avoid surface charging and consequent damage to the sample a low accelerating voltage (1 kV) with typical vacuum pressure of 10⁻⁵ mbar at a working distance of 3 mm was applied. An Everhart–Thornley detector (ETD) for low magnification images and a through lens detector (TLD) for high magnification images were used for the collection of SE images.

The specimens were rinsed twice with 70% ethanol using an ultrasonic cleaner (KQ118, Kunshan, China), twenty seconds for each time. And then, samples were dehydrated in agraded series of 80%, 90% ethanol for 20 min each and 99.9% ethanol for 30 min twice before being transferred to a mixed solution of ethanol and tertbutanol (3:1, 1:1, and 1:3, by volume) for 15 min each, and finally dipped into 100% tert-butanol for 30 min. After that, the samples were dried with a freeze-drier (VFD-21S, SHINKKUVD, Japan) for 3 h. The dried antennae were then separated from bodies and mounted on the aluminum stubs under a stereomicroscope with double-sided copper sticky tape and coated with gold/palladium (40/60) in a high-resolution sputter coater (MSP-1S, Hitachi, Tokyo, Japan). Fine morphology of four female and four male antennae were examined with a Nova Nano 450 SEM (FEI, America) operated at 15 kV.

The FEI Nova Nano 450 SEM is provided with a through lens detector which includes a voltage controlled deflector electrode. The deflector electrode channels the signal into the SE detector. The deflector electrode is set to predetermined deflector voltages and an image is generated for each deflector voltage. Spectra and hyperspectral images are acquired through post-processing of such image series. Stage bias experiments have been performed to allow energy calibration of this process through experiments. Fiji software was utilised to perform image processing and extract S-curves from the SEHI stacks. Histogram and spectral off-set normalisation were applied retrospectively to optimise all images for brightness and contrast, and to limit the effects of differing sample work functions. Once S-curves were obtained they are differentiated to produce the SE spectra. Component analysis was then preformed to image stacks utilising non-negative matrix factorization (nnmf) to isolate components of interest.