Jsm 5200 scanning electron microscope

Chloroauric acid (HAuCl4), Bovine serum albumin (BSA), sucrose (C12H22O11), and potassium carbonate (K2CO3, ≥ 99.0%), skimmed milk, were all purchased from Sigma Aldrich, MO, USA. Recombinant human coronavirus SARS-CoV-2 spike glycoprotein S1 (ab 288546), Recombinant anti-SARS-CoV-2 antibodies (ab 281311), Recombinant angiotensin-converting enzyme 2 (ACE2) (ab151852), all from Abcam, Cambridge, UK. SARS-CoV recombinant protein (MBS569928), MERS CoV spike S1 (MBS434229) antigen from MyBioSource, California, US, SARS-CoV-2 S1 nanobodies (AssayGenie, Dublin, Irland), Pierce DAB substrate kit (cat. no. 34002, Thermo Fisher USA). A sample pad and absorption pad (cat no CFSP173000), a glass fiber conjugate pad (cat no. GFDX103000), high flow nitrocellulose membrane (NC) (cat no HF09002XSS) were purchased from Merch Millipore (Darmstadt, Germany). UV-Vis-spectrophotometer (Thermofisher-USA). Manual dispenser (Nanomat 4-CAMAG-laborto), PH meter (Jenway 3510, UK), High-speed centrifuge (Eppendorf, 5430R, Germany). Ultrapure water used throughout was generated from a Millipore Milli-Q water purification system (Billerica, MA, USA). Gel documentation system (Gel Doc XR+) (Biorad, USA) & data analysis by “Image lab” software. JEOL JSM5200 Scanning Electron Microscope, Japan.

The JSM-5200 scanning electron microscope (Jeol Inc., Tokyo, Japan) was used to examine the morphology of microspheres. They were sprinkled over a double-sided adhesive tape mounted on a metal specimen stub. Prior to examination with SEM, microspheres were coated by using a SC7620 sputter coater (VG Microtech, West Sussex, UK).

SEM was performed as described in^{23 (link)}. Briefly, Ectocarpus gametes were released and grew on a polycarbonate filtration membrane (Nuclepore, diameter 13 mm, Cat N° 110406 N, Whatman). After two weeks, parthenosporophytes were fixed in seawater with 3% paraformaldehyde for 1 h and then washed 10 min in a diluted Artificial Sea Water (ASW) ASW: H₂O (3:2), ASW: H₂O (2:3) and H₂O, followed by successively dehydration steps in 30%, 50%, 70%, 90%, 2 times 95% and 3 times 100% EtOH. They were finally dried using a critical point dryer (Baltec CPD 030, Balzer), covered with a 25 nm thick gold layer and observed with a JEOL JSM 5200 scanning electron microscope.

Microspheres were mounted onto a double adhesive tape attached to a metal stub. The microspheres were sputtered with gold by using a SC7620 sputter coater (VG Microtech, West Sussex, UK). Their morphology was observed by the JSM-5200 scanning electron microscope (Jeol Inc., Tokyo, Japan).

Specimens were photographed using an Olympus Camedia E‐520 camera attached to an Olympus SZX16 stereomicroscope or to the eye piece of an Olympus BH2 transmission microscope, and a SEM JEOL JSM-5200 scanning electron microscope at the Zoological Museum of the University of Turku. Digital images were prepared using CombineZP image stacking software. Illustrations of internal genitalia were made after clearing them in a 10% KOH aqueous solution. Lengths of leg segments were measured on the dorsal side. Measurements of legs are listed as: total length (femur, patella, tibia, metatarsus, tarsus). All measurements are given in millimetres.

P. rhoeas flowers were collected at the Botanical Garden “Angelo Rambelli” of Tuscia University, Viterbo, and the flowers processed in our laboratory under observation with a stereo microscope. Petals and stamens were removed and the pistils excised using a razor blade. The ovaries and their ovule content were observed using a stereo microscope and some images were taken. Samples were then processed for SEM investigation.
For SEM observations, ovaries containing ovules were collected in tubes, washed with phosphate buffered saline, and fixed with 4% paraformaldehyde and 5% glutaraldehyde, pH 7.2 in 0.1 M cacodylate buffer for 1 h at 4 °C [26 (link)]. After rinsing overnight in the same buffer, samples were post-fixed in 1% osmium tetroxide in cacodylate buffer for 1 h at 4 °C. After two washings in the same buffer, samples were dehydrated in a graded ethanol series. Ovaries containing ovules were dried using the critical point drying method with CO₂ in a Balzers Union Critical Point Dryer (CPD) 020, sputter-coated with gold in a Balzers MED 010 unit and observed with a JEOL JSM 5200 Scanning Electron Microscope.