Liver was fixed overnight in 4% paraformaldehyde in 0.1 M phosphate-buffered saline (PBS). After being embedded in paraffin, the specimens were cut cross-sectionally into 4-µm sections. All sections were incubated in sodium citrate solution (pH 6.0) at 95 °C for 40 min and then blocked with 5% fetal bovine serum in PBS containing 0.1% Tween-20 at room temperature for 2 h. Sections were then incubated with anti-CDKN2A/p16Ink4a antibody (catalog no. 54210; Abcam, Waltham, MA, USA) with 1:250 dilution at 4 °C for overnight in moist chamber. They were then washed and incubated with goat anti-mouse IgG (catalog no. ab150116; Abcam, Waltham, MA, USA) with 1:500 dilution at room temperature for 1 h. Vectashield Plus antifade mounting medium (catalog no. H-1900; Vector Laboratories, Newark, CA, USA) containing DAPI at 1:5,000 dilution was applied, and fluorescent signals were visualized under a fluorescent microscope (Ni-U model eclipse, Nikon, Japan).
Model eclipse ni u
Manufactured by Nikon
Sourced in Japan
The Nikon ECLIPSE NI-U is a biological microscope designed for laboratory use. It features a sturdy, ergonomic design and provides high-quality optical performance. The microscope is equipped with a range of illumination methods, including transmitted and reflected light, to accommodate various specimen types. The ECLIPSE NI-U is suitable for a variety of applications in the life sciences and research fields.
Automatically generated - may contain errors
Lab products found in correlation
Vectashield plus antifade mounting medium, by Vector Laboratories (1 mentions)
Anti cdkn2a p16ink4a antibody, by Abcam (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Cellsens image acquisition software, by Olympus (1 mentions)
Dylight 594, by Vector Laboratories (1 mentions)
Diamond antifade mounting medium, by Thermo Fisher Scientific (1 mentions)
Prism software, by GraphPad (1 mentions)
5 protocols using model eclipse ni u
1
Immunohistochemical Analysis of p16Ink4a Expression
2
Histological Analysis of Aortic Tissue
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After euthanasia, the aorta was collected and washed with ice-cold sterile normal saline solution, fixed in 4% paraformaldehyde for 12 h at 4 °C. Tissues were dehydrated and cleared by graded ethanol and xylene, respectively, and embedded in paraffin. Paraffin blocks were cut into 5-µm thickness and stained with hematoxylin and eosin (H & E) for histological examination. Slides were visualized under a light microscope (Ni-U model eclipse; Nikon, Tokyo, Japan) with cellSens image acquisition software (Olympus, Tokyo, Japan).
3
Quantifying Mycorrhizal Colonization in Plants
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Plants were harvested on July 30, 2017 when they reached the highest plant biomass. After harvest, we measured aboveground plant biomass of each plant, and root N concentration (Vario EL CUBE, Germany) and mycorrhizal colonization rate for native plants. Mycorrhizal colonization was determined following previous procedures (Trouvelot et al., 1986 ; Liu et al., 2015b (link)). Briefly, 50 randomly selected root segments were washed with distilled water and then soaked in 10% KOH solution for 50 min at 90 °C. Afterwards, the roots were soaked in 5% HCl solution for 5 min and then washed with distilled water. The roots were stained in 0.05% acidic magenta solution, flattened on a slide, and then examined for mycorrhizal fungi colonization using a 200 × microscope (Nikon MODEL ECLIPSE NI-U, Japan). The roots were classified into five categories of mycorrhizal colonization, each assigned with a weight of 0.95, 0.70, 0.30, 0.05 and 0.01. The final colonization rate of the native roots was calculated as follows: where are the number of root segments belonging to each of the five categories.
4
Quantifying Duodenal PMCA1 Expression
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Duodenal segments were fixed overnight in 0.1 M phosphate-buffered saline (PBS) containing 4% paraformaldehyde. After being embedded in paraffin, the specimens were cut longitudinally into 4-µm sections which were later incubated at 95 °C for 20 min in sodium citrate buffer solution (pH 6.0). Non-specific bindings were blocked for 2 h by 4% bovine serum albumin, 5% normal goat serum, and 0.1% Tween-20 in PBS. Thereafter, sections were incubated at 4 °C overnight in moist chamber with rabbit monoclonal anti-PMCA1 (plasma membrane Ca2+ ATPase) at 1:500 dilution (Cat# ab190355, Abcam, RRID:AB_2893200). After being washed with 0.5% Tween-20 in PBS, sections were incubated for 1 h at room temperature in the dark with anti-rabbit Dylight 594 (Cat#DI-1594, Vector Laboratories) in blocking buffer. Sections were mounted with slow fade Diamond antifade mounting medium containing DAPI (Cat# S36964, Invitrogen) visualized under a fluorescent microscope (model eclipse Ni-U; Nikon). Fluorescent intensity was measured from 5 images per rat, 3–4 villi per image (total of 15–20 villi/rat), 5 rats/group by NIS-elements BR imaging software (version 4.0).
5
Automated Microplastic Quantification via YOLO v5
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In this study, six different types of MP sheets are applied: PC, PET, PVC, PP, PS, and PMMA. These samples with varying birefringence values are purchased from Xinsheng Plastic Material Company, China. To ensure sample diversity, additional materials such as glass, lens paper, and algae are included in the control group.
The samples are imaged using a stereo microscope (Nikon Model Eclipse Ni-U, Japan) with a digital camera. The captured MP images by the polarization CMOS camera are accurately labeled using ImageJ software (National Institutes of Health, US), and the dataset is divided into training, validation, and test sets at a ratio of 8:1:1. The YOLO v5 model, a lightweight CNN, is combined with a Strong-SORT network to segment, measure, and count the MP samples. The DL model is further fine-tuned. To fine-tune the hyperparameters, specifically the learning rate, batch size, and whether augmentations should be used, a manual grid search is performed. The area and MFD values are calculated by a self-developed Python script. The scatter diagram and line chart are visualized by GraphPad Prism software (GraphPad Inc, USA).
The samples are imaged using a stereo microscope (Nikon Model Eclipse Ni-U, Japan) with a digital camera. The captured MP images by the polarization CMOS camera are accurately labeled using ImageJ software (National Institutes of Health, US), and the dataset is divided into training, validation, and test sets at a ratio of 8:1:1. The YOLO v5 model, a lightweight CNN, is combined with a Strong-SORT network to segment, measure, and count the MP samples. The DL model is further fine-tuned. To fine-tune the hyperparameters, specifically the learning rate, batch size, and whether augmentations should be used, a manual grid search is performed. The area and MFD values are calculated by a self-developed Python script. The scatter diagram and line chart are visualized by GraphPad Prism software (GraphPad Inc, USA).
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