For PAS staining, conjunctival tissue was fixed in 4% paraformaldehyde overnight and embedded in paraffin. Serial sections, 6-μm thick, were cut from each sample. The sections were deparaffinized and stained with the 0.5% PAS for identification of goblet cells. Sections from each group were photographed with a microscope equipped with a digital camera. The number of positively stained goblet cells in the superior and inferior conjunctiva was counted in 3 sections from each eye by using image analysis software (Media Cybernetics, Silver Spring, MD). Data are presented as the average number of goblet cells per millimeter.
Image analysis software
Manufactured by Media Cybernetics
Sourced in United States
Image analysis software is a computer program designed to process and analyze digital images. It provides tools for users to perform various image processing tasks, such as enhancing, segmenting, and extracting information from images.
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Lab products found in correlation
8 protocols using image analysis software
1
Conjunctival Goblet Cell Quantification
2
Femoral Head Protein Analysis and cGMP/PKG Quantification
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Total protein was isolated by crushing and homogenizing the femoral head with RIPA buffer (Thermo Scientific, USA). Proteins were separated and transferred to membranes following standard protocols and probed using mouse monoclonal anti-VEGF (1:500; Abcam, USA) antibody. Protein expression was visualized on a film with an enhanced chemiluminescence kit (NEN Life Science Products Inc., Boston, MA, USA). Relative protein expression was determined using image analysis software (Media Cybernetics). For the quantitative analysis of protein, the intensity of each test band was expressed as the optical density (OD). β-actin was detected by mouse monoclonal anti-actin antibody (1:1000; Santa Cruz, USA) and was used as an internal control.
Measurement of Tissue cGMP and PKG Activity. Bone tissue cGMP level was measured by enzyme-linked immunosorbent assay using cGMP Enzyme Immunoassay Kit (R&D systems, Minneapolis, MN) according to the manufacturer's directions. PKG activity was measured according to the manufacturer's instructions (Cyclex, Nagano, Japan). Spectrophotometric absorbance was measured at 450 nm, and the results were normalized per mg of protein.
Measurement of Tissue cGMP and PKG Activity. Bone tissue cGMP level was measured by enzyme-linked immunosorbent assay using cGMP Enzyme Immunoassay Kit (R&D systems, Minneapolis, MN) according to the manufacturer's directions. PKG activity was measured according to the manufacturer's instructions (Cyclex, Nagano, Japan). Spectrophotometric absorbance was measured at 450 nm, and the results were normalized per mg of protein.
3
Conjunctival Goblet Cell Quantification
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The eye and adnexa were surgically excised, fixed in 4% paraformaldehyde, and embedded in paraffin. Six-micron sections were stained with PAS reagent. Sections obtained from four animals in each group were examined and photographed using a microscope (Olympus Corp., Tokyo, Japan) equipped with a digital camera. Goblet cell density in the superior and inferior conjunctiva was measured in three sections from each eye using image analysis software (Media Cybernetics, Silver Spring, MD, USA) and expressed as the number of goblet cells per 100 μm.
4
Wound Healing Assay of Transfected Cells
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The transfected SK-Hep1 cells from different groups were seeded into 6-well plates. A sterile 200-μL pipette tip was used for creating linear scratches on cell monolayers. Images of wounded areas were photographed at 0 and 24 hrs after the scratch and analyzed by Image analysis software (Media Cybernetics, MD, USA).
5
Quantifying Conjunctival Goblet Cells
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Eyes and adnexa were surgically excised, fixed in 4% paraform-aldehyde, and embedded in paraffin. Six-micrometer sections were stained with Periodic acid-Schiff reagent. Sections from each group were examined and photographed with a microscope (BX53; Olympus, Tokyo, Japan) equipped with a digital camera (F2; Foculus, Finning, Germany). Goblet cells in the conjunctiva were counted in three sections from each eye using image analysis software (Media Cybernetics, Silver Spring, MD, USA) and expressed as the number of goblet cells per 100 µm [18 (link),19 (link)].
6
Histological Evaluation of Achilles Tendon
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Harvested Achilles tendon specimens were fixed in 4% paraformaldehyde for 24 h and decalcified for 7 days using 10% ethylenediaminetetraacetic acid (pH 7.0). The specimens were then paraffin‐embedded and sectioned with a paraffin microtome (RM2125 RTS, Leica) to obtain 4 μm thick pathological sections. Haematoxylin and eosin (HE) staining and immunofluorescence staining were performed according to protocol and guidelines. Histological images were acquired by a scanning tissue microscope (Olympus BX51) and Image analysis software (Media Cybernetics Inc) was used to track and quantify HO areas based on the stained images.
7
Conjunctival Goblet Cell Density Analysis
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Eyes were surgically excised, washed immediately with ice-cold saline to remove as much blood as possible, fixed in 4% paraformaldehyde, and embedded in paraffin. Next, six-micrometer conjunctival sections were stained with periodic acid-Schiff (PAS) reagent. Sections were examined and photographed with a microscope (BX53; Olympus, Tokyo, Japan) equipped with a digital camera (F2; Foculus, Finning, Germany). Goblet cell density in the superior and inferior conjunctiva was measured in three sections from each eye using image analysis software (Media Cybernetics, Silver Spring MD) and expressed as the number of goblet cells per 100 μm.
8
Conjunctival Goblet Cell Analysis
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Conjunctival tissue was surgically excised, fixed in 4% paraformaldehyde, and embedded in paraffin. We stained 6lm sections with periodic acid-Schiff (PAS) reagent. Sections from four animals from each group were examined and photographed with a microscope (Olympus Corp., Tokyo, Japan) equipped with a digital camera. Goblet cell density in the superior and inferior conjunctiva was measured in three sections from each eye using image analysis software (Media Cybernetics, Silver Spring, MD, USA) and was expressed as the number of goblet cells per 100 lm.
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