Formalin-fixed tissue specimens were embedded in paraffin. Tissue sections (5-μm thick) were deparaffinized in xylol (534056; Sigma-Aldrich) and rehydrated in serial alcohols and were later used for hematoxylin-eosin (HE) (hematoxylin, 104302; eosin, 115935; Merck KGaA, Darmstadt, Germany) staining and double immunostaining (DakoCytomation EnVision®, Doublestain System [HRP-AP], Dako Inc., Carpinteria, CA, USA). Digital images of HE and immunostained sections were made on a Photomicroscope Olympus (Olympus BX41TF; Olympus, Tokyo, Japan) with a digital camera (Olympus C5060; Olympus) for the acquisition and analysis of the images.
C 5060
Manufactured by Olympus
Sourced in Japan, United States
The C-5060 is a digital camera designed for laboratory and scientific applications. It features a 5 megapixel CCD sensor and a high-quality optical system for capturing detailed images. The camera provides connectivity options for integration with various laboratory equipment.
Automatically generated - may contain errors
Lab products found in correlation
Ckx41, by Olympus (3 mentions)
Th4 200, by Olympus (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
M165 fc, by Leica (2 mentions)
Xylol, by Merck Group (1 mentions)
Matrigel, by Solarbio (1 mentions)
Cetylpyridinium chloride solution, by Merck Group (1 mentions)
Synergy h1, by Synergy Software (1 mentions)
Alizarin red, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Szx12 microscope, by Olympus (1 mentions)
Dmlb microscope, by Leica (1 mentions)
24 well culture plate, by Corning (1 mentions)
6 well culture plate, by Corning (1 mentions)
Cultrex matrix, by Bio-Techne (1 mentions)
Ck2 inverted microscope, by Olympus (1 mentions)
Uc6 ultra microtome, by JEOL (1 mentions)
Nanodrop, by Eppendorf (1 mentions)
Histostain plus kits, by Bioss Antibodies (1 mentions)
Vascular endothelial growth factor (vegf), by Bioss Antibodies (1 mentions)
41 protocols using c 5060
1
Paraffin-Embedded Tissue Histology Analysis
2
Histological Evaluation of Kidney Injury
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For light microscopic evaluations, the kidney samples were fixed in 10% buffered formalin for 48 h and processed for routine paraffin embedding. For general morphological evaluation, approximately 5-µm-thick sections were stained with hematoxylin and eosin (H&E). All of the stained sections were observed and photographed with a digital camera (Olympus C-5060; Olympus, Japan) attached to a photomicroscope (Olympus BX51; Olympus, Japan).
The microscopic scoring of the right kidneys was carried out in a blinded fashion by a histologist who was unaware of the treatment groups and assigned a score that represented the approximate extent of hemorrhage, tubular epithelial cell damage, and necrosis according to Abdelrahman et al. [13]. These parameters were evaluated on a scale of 0–3, which ranged from absent (0) and mild (1) to moderate (2) and severe (3) (Table 2).
The microscopic scoring of the right kidneys was carried out in a blinded fashion by a histologist who was unaware of the treatment groups and assigned a score that represented the approximate extent of hemorrhage, tubular epithelial cell damage, and necrosis according to Abdelrahman et al. [13]. These parameters were evaluated on a scale of 0–3, which ranged from absent (0) and mild (1) to moderate (2) and severe (3) (Table 2).
3
In Vitro Tube Formation Assay
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96-Well plates were pre-chilled. Each well was coated with 50 µl of 50% Matrigel (Solarbio, M8370, China), which was polymerized for 30 min at 37 °C. HUVECs were harvested from each group (3 × 104 cells/well) in 100 µl DMEM medium supplemented with 1% FBS. These aliquots of cell cultures were added to each well and incubated at 37 °C, 5% CO2 for 12 h. Tube formation was observed using a light microscope (×100 magnification) and photographed with a digital camera (Olympus C-5060, Japan).
4
Alizarin Red Staining for Osteoblast Mineralization
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alizarin red staining was used to evaluate the mineralized nodule formation on the final days of treatment (days 6 and 13). This assay was based on quantifying the mineralized matrix produced by osteoblast-like cells. The treatments were aspirated, and the cells were fixed in 70% cold ethanol for 2 h. The ethanol was then aspirated, and each well received 100 µL of alizarin red (pH 4.2; Sigma-Aldrich), after which the samples were incubated under continuous shaking for 20 min. The wells were rinsed twice with deionized water, and incubated at room temperature to dry for 24 h. Next, mineral nodules of each group were identified and photographed by using a light microscope (Olympus BX51; Olympus, Miami, USA) equipped with a digital camera (Olympus C5060; Olympus, Miami, USA). Lastly, 150 µL of cetylpyridinium chloride solution (Sigma-Aldrich) was added to each well, and maintained under continuous shaking for 15 min to dissolve the nodules. The quantitative analysis was performed based on the mineralized nodule formation measured by absorbance at 570 nm in a spectrophotometer (Synergy H1). The mean absorbance value of the control group (Control DMEM) on day 6 was considered as 100% of mineralized matrix deposition. 18 (link)
5
Beating Characteristics of Differentiated Embryoid Bodies
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The beating characteristics of differentiated EBs were determined using CBAnalyser (for more details, see [19 (link)]). Briefly, the digital video of beating EBs or individual cells was recorded in phase contrast for approximately 5 minutes per sample using a digital camera (Olympus C-5060) mounted on an inverted microscope (Olympus IX41) at room temperature. The beating parameters were then analyzed from the recordings by CBAnalyser. Representative videos documenting the beating of HIF1alpha+/+ and HIF1alpha-/- mESC-CMs (S1 Mov and S2 Mov ), together with figures showing the analysis performed by CBAnalyser (S2 Fig ) are included in the Supporting information files.
6
Cell Adherence Quantification Assay
7
Cytogenetic Analysis of Allium cepa Root Tips
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Root tips of (1–1.5 cm) A. cepa bulbs were used for cytogenetic analysis. Root tips were pretreated with saturated para-dichlorobenzene solution for 4 h and then fixed for a day in solution of ethanol–acetic acid (75–25%). Fixated tips were hydrolyzed using 1 N HCl at 60 °C for 17 min before stained with Feulgen for 1.5–2 h. To prepare squash slides, stained root tips were crushed with a drop of 45% acetic acid (Sharma and Gupta 1982 ). Mitotic index (MI), frequencies of micronucleus (MN), and chromosomal abnormalities (CAs) were observed at 500 × magnification with a microscope (Olympus CX41) and photographed with a digital camera (Olympus C-5060). MI was calculated by analyzing 1,000 of cells from ten slides prepared from each treatment (10,000 for each treatment). CAs and MN frequencies of each treatment were observed on ten slides (1,000 cells for each treatment) (Çavuşoğlu et al. 2021 ).
8
Anatomical Alterations in Root Tip Meristems Under HgCl2 Exposure
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Root tips were investigated for HgCI2-induced anatomical alterations in meristematic cells. Before cross-sections were taken manually, root tips were thoroughly rinsed with distilled water to remove residues. Cross-sections were stained with a drop of 1% methylene blue. Meristematic damages were observed at 500 × magnification with a light microscope (Olympus CX41 with Olympus C-5060 digital camera). Meristematic cell damages were classified as no damage, little damage, moderate damage, and severe damage (Yalçın et al. 2021 ).
9
Meristematic Cell Damage by UV Exposure
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Meristematic cell damages caused by UV applications were investigated using the cross-sections of decapitated roots. Each root tip was taken from 10 randomly selected bulbs from each group. 1 ml of methylene blue (1%) was utilized to stain the cross-sections. Three slides for each bulb were analyzed for meristematic cell integrity under a research microscope (Olympus CX41) at 500 × magnification. A digital camera (Olympus C-5060) was utilized to photograph the slides. The severity of the damages was classified as “no damage,” “light damage,” “moderate damage,” and “intense damage.”
10
Wound Healing Assay with CNP and UVA
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The L929 cells were treated with 100 nM of CNP for 24 h and irradiated with UVA (30 J/cm2). Immediately after irradiation, a sterile 200 μL pipette tip was used to make a straight scratch on the monolayer of cells attached. The pictures were taken at 0, 24, and 48 h after the scratch. Wound repopulation was assessed with a light microscope (Olympus BX51, Miami, FL, United States) equipped with a digital camera (Olympus C5060, Miami, FL, United States). Photomicrographs were taken at 5× magnification and cell proliferation area was measured using Image-J 1.45S software (Wayne Rasband, National Institutes of Health, Bethesda, MD, United States).
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