The surface morphology of albumin, and albumin-SNAP, was evaluated using optical microscopy and atomic force microscopy. Samples were viewed using an Olympus BX50 optical microscope with a camera at 20× magnification. Samples were also measured by a Bruker atomic force microscope (Dimension Icon or Multimode) with ScanAsyst at 40 μm scan size.
Bx50 optical microscope
Manufactured by Olympus
Sourced in Japan
The Olympus BX50 is an optical microscope designed for a variety of laboratory applications. It features a sturdy, ergonomic design and provides high-quality, clear images through its optical system. The BX50 is capable of various magnification levels to accommodate different sample types and examination needs.
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Lab products found in correlation
Dimension icon, by Bruker (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
Image pro plus 4, by Media Cybernetics (1 mentions)
Axio scan z1 slide scanner, by Zeiss (1 mentions)
Zen blue edition software, by Zeiss (1 mentions)
Expose rabbit specific hrp dab detection ihc kit, by Abcam (1 mentions)
Image pro plus version 6, by Media Cybernetics (1 mentions)
He staining kit, by Solarbio (1 mentions)
Szx12 stereomicroscope, by Olympus (1 mentions)
Microtome, by Leica (1 mentions)
Cell a software, by Olympus (1 mentions)
Dp71 camera, by Olympus (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Sytox green nucleic acid stain, by Thermo Fisher Scientific (1 mentions)
Bx41 epifluorescence microscope, by Olympus (1 mentions)
18 protocols using bx50 optical microscope
1
Characterizing Albumin Surface Morphology
2
Bead Hydration and Morphology Analysis
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The lyophilized beads were hydrated with 1.5 mL purified water and left for 30 minutes to hydrate. The volume of beads was determined by transferring them to a 5 mL measuring cylinder and reading the sedimented volume. Bead morphology was observed under an Olympus BX50 optical microscope at X 10 magnification. The diameter of 200 beads per vial was measured.
3
Quantitative Bone Regeneration Analysis
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The specimens were fixed in 10% formalin for six weeks before being washed and demineralized in 2.5% NaOCl and 17% EDTA solution for 18 days. After embedding, the specimens were sliced to create sections of 3–4 μm thickness. The slices were then cut, along the sagittal plane, passing through the center of the bone defect in the anterior and posterior directions. The specimens were subjected to hematoxylin-eosin (H and E), Russell–Movat pentachrome staining (American MasterTech, USA), and tissue slides were prepared. The specimens were photographed at 12.5× and 40× magnifications using a BX50 optical microscope (Olympus, Tokyo, Japan). The new bone, remaining bone graft material (RB), fibrovascular connective tissue (FCT), and shrinkage volume (SV) (which is the unfilled volume of the defect) were measured within the defect area using Image-Pro Plus software (Media Cybernetics, Silver Spring, MD, USA). Each area was described as a percentage of the total defect area.
4
COVID-19 Immunohistochemical Analysis
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The slides immunolabeled with anti-IL-33, anti-CASP-1, anti-B1R, anti-B2R and anti-ACE2 were scanned with the assistance of the Axio Scan Z1 slide scanner (Zeiss, Jena, Germany) and submitted to the generation of 30 HPF (COVID-19 group) and 20 HPF (H1N1 and CONTROL groups) by the ZEN Blue Edition software (Zeiss, Jena, Germany). Analyses were performed blindly by an observer. The areas of immunoexpression were quantified using Image Pro-Plus 4.5 software (Media Cybernetics, Rockville, MD, USA), and subsequently, these areas were converted to percentages. Data were subjected to statistical analysis.
The slides immunolabeled with anti-IgE and anti-Tryptase were observed exclusively in the alveolar septum and perivascular spaces by counting immunostained MCs in 20 randomized HPF (40X, Olympus Objective, 0.26 mm2 per sample), using a BX50 optical microscope (OLYMPUS, Tokyo, Japan). Average scores were obtained by screening 20 randomized HPFs (total area of 5.2 mm2 per case). Data were subjected to statistical analysis.
The slides immunolabeled with anti-IgE and anti-Tryptase were observed exclusively in the alveolar septum and perivascular spaces by counting immunostained MCs in 20 randomized HPF (40X, Olympus Objective, 0.26 mm2 per sample), using a BX50 optical microscope (OLYMPUS, Tokyo, Japan). Average scores were obtained by screening 20 randomized HPFs (total area of 5.2 mm2 per case). Data were subjected to statistical analysis.
5
Immunohistochemical Analysis of Twist1 in Lung
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Sagittal sections of the right lung were placed in 4% paraformaldehyde and processed for paraffin embedding. Sections (5 μm) were prepared and mounted on glass slides prior to overnight incubation at 4°C with anti-Twist1 antibody (1:500). Slides were washed and incubated with corresponding secondary antibodies conjugated with alkaline phosphatase (Expose Rabbit specific HRP/DAB detection IHC kit; ab80437; Abcam). Sections were evaluated using an Olympus BX50 optical microscope (Olympus Corporation, Tokyo, Japan) equipped with an image analysis program (Image Pro Plus version 6.0; Media Cybernetics, Inc., Rockville, MD, USA).
6
Direct Planktonic Cell Counting
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The density of planktonic cells was quantified by direct cell counting as described previously (Igarashi et al., 2020 (link)). Briefly, during cultivation, 100 μL of culture was periodically sampled with a disposable syringe. The sample was mixed with a LIVE/DEAD BacLightTM Bacterial Viability Kit and then cells were directly counted using a bacterial counting chamber and a BX50 optical microscope (Olympus, Tokyo, Japan).
7
Assessing Sea Urchin Embryo Development
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Exposed and control P. lividus animals were sacrificed and gonads were gently washed to allow gametes collection. In particular, for each experimental condition, eggs from treated females (3 females/treatment/experiment) were pooled and maintained in ASW until fertilisation with sperms (3 males/treatment/experiment) collected and diluted (1:500) in ASW. Similarly, pooled eggs from three untreated animals were fertilised in presence of pooled sperms from control males. All the performed fertilisation occurred in the condition of sperms/eggs ratio corresponding to 100:1. Eggs were rinsed after fertilization. At least 6000 embryos were allowed to develop in 250 mL tanks at 18 °C under gentle agitation in ASW and they were examined until the pluteus stage (48 hpf). The percentages of embryos showing normal or abnormal phenotype were determined by counting about 100 embryos/replicate in each treatment using the Olympus BX50 optical microscope.
8
Detailed H&E Staining Procedure
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An H&E staining kit (G1120, Solarbio, Beijing, China) was used to stain the tissue sections. The sections were deparaffinized in xylene twice (2–5 min each time), and rehydrated with successive washes in 100% ethanol for 1 min, 95% ethanol for 1 min, 95% ethanol for 2 min, 75% ethanol for 1 min, tap water for 2 min, and distilled water for 2 min. The tissue sections were then stained with hematoxylin for 5 min, rinsed with distilled water for 1 min, differentiated with 75% hydrochloric acid ethanol for 30 s, rinsed with 50°C tap water for 5 min, rinsed with distilled water for 5 min, then stained with eosin for 1–2 min, and rinsed again with distilled water. Furthermore, sections were dehydration with graded alcohol: 95% ethanol (2×1 min), 100% ethanol (2×1 min), and clearing in xylene (2×1 min). The mounted slides were then examined and photographed using an Olympus BX50 optical microscope (Olympus, Tokyo, Japan) with an ISCapture microscope imaging system (Tucsen Photonics). The staining intensity was analyzed by Image-Pro Plus 5.0 software and expressed as an integrated optical density value. Ten fields were randomly selected at 100× magnification for each section.
9
Paralarvae Morphological Analysis
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Morphological analysis was carried out using an Olympus BX50 optical microscope. Live paralarvae were sampled and anaesthetized with 0.35 mM MgCl2, 1 : 2 diluted with seawater. A number of 50 paralarvae per treatment were scored for mortality. Measurements of mantle (ML) and arm length (AL) were taken in both dorsal and ventral side of paralarvae. Images were analysed using NIH-Image public domain software (version 1.61 ).
10
Growth Dynamics and Cellular Analysis
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The growth period was divided into three stages. The first growth phase was from sprouting to the development of unifoliolate leaves. The secondary growth phase was from the development of the unifoliolate leaves to that of the first trifoliolate leaves. The third growth phase was from the development of the first trifoliolate leaves to the reproductive R3 stage (1 cm-sized pod). The size of the unifoliolate leaves of the mutant lines was measured, and leaf area was determined as leaf size multiplied by the length and width of the unifoliolate leaf. SPAD values of the first trifoliolate leaves were measured using a chlorophyll analyzer every 2 days from 14 to 36 days after flowering (DAF). The cell size of the unifoliolate leaves was determined. Briefly, manicure was applied to the surface of unifoliolate leaves and allowed to dry. Then, the epidermal cells in the leaves were peeled off along with the manicure and observed under the BX50 optical microscope (Olympus, Tokyo, Japan). The cell size of the epidermal cells in the microscopic images was measured using ImageJ software (https://imagej.nih.gov/ij/index.html).
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