H1R protein expression in the atheroma was assessed in histological sections by using anti-H1R antibody (Alomone Labs, Israel) followed by Alexafluor-488 staining for immunofluorescence detection. Nucleus was counterstained with DAPI. Images were captured and analyzed using Leica TCS SPE Confocal Microscope as well as a Nikon 80i fluorescent microscope. Integrated density of H1R fluorescence was measured using Adobe Photoshop CC, Adobe Corporation.
80i fluorescent microscope
Manufactured by Nikon
Sourced in Japan
The Nikon 80i fluorescent microscope is a high-performance laboratory instrument designed for fluorescent imaging and analysis. It features advanced optics and illumination systems to provide clear and detailed images of fluorescently labeled samples.
Automatically generated - may contain errors
Lab products found in correlation
Extravidin cy3, by Merck Group (2 mentions)
Image it fx signal enhancer, by Thermo Fisher Scientific (2 mentions)
Elements ar software, by Nikon (2 mentions)
Cm1950, by Leica (2 mentions)
Tcs spe confocal microscope, by Nikon (1 mentions)
Ds qi1mc monochrome camera, by Nikon (1 mentions)
8 m pore inserts, by BD (1 mentions)
24 well companion plate, by BD (1 mentions)
Lyticase enzyme, by Merck Group (1 mentions)
Alexa 488 secondary antibody, by Thermo Fisher Scientific (1 mentions)
Rm2245, by Leica (1 mentions)
Ethidium homodimer 1, by Thermo Fisher Scientific (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
8 well chamber slide, by Thermo Fisher Scientific (1 mentions)
Metamorph digital imaging system, by Nikon (1 mentions)
Stereo investigator software, by MBF Biosciences (1 mentions)
Microfire digital camera, by Optronics (1 mentions)
Ds ri2 camera, by Nikon (1 mentions)
Nis elements ar imaging software, by Nikon (1 mentions)
Dapi 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions)
19 protocols using 80i fluorescent microscope
1
Histological Assessment of H1R Expression
2
Quantifying New Cell Density
3
Myotube Staining and Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stained for MyHC using MyHC polyclonal rabbit anti-mice antibody (sc-20,641, 1:150, Santa Cruze) followed by anti-rabbit TRITC or FITC-labeled secondary antibody (Jackson Lab, 1:400, USA). DAPI was used to stain nuclei. Myotube was defined as 3+ nucleuses within a cellular structure in order to rule out myoblast cells undergoing mitosis. The images of five locations including up, down, left, right sides, and center of each slide were photographed using an 80i Nikon fluorescent microscope (Nikon, Japan). A total of 30 images/group within six repeats were taken by using the same imaging parameters. Images were analyzed by two pathologists using Image J (Java) software (National Institutes of Health, USA) in a double-blind manner. Morphology was assessed by myotube length, number of myotube per view, and number of myoblast fusion (nuclei numbers as the indicator) per myotube [15 (link)–17 (link)]. In order to facilitate the description of myotube characteristics, the myotubes were divided into 3 types including short, medium, and long myotubes. Short myotube was defined as those shorter than 200 μm with less than 5 myoblast fusions, and long myotubes were defined as those longer than 400 μm with more than 10 myoblast fusion; medium myotube were those between the short and long myotubes.
4
Quantitative analysis of microglial markers in Alzheimer's disease
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analysis was performed on sections immunostained against Iba1 or CD68 antigen and counterstained with Th-S. Microphotographs of the brain cortex were taken at × 20 objective magnification under Tetramethylrhodamine (for Iba1/Cy3 and CD68/Cy3) and Fluorescein isothiocyanate (for Th-S) channels of 80i Nikon fluorescent microscope (Nikon Corp. Tokyo, Japan) using DS-Qi1Mc monochrome camera (Nikon Corp. Tokyo, Japan). The load of Iba1 and CD68 positive cells (defined as the percentage of a microphotograph area occupied by anti-Iba1 or anti-CD68 positive immunostaining) was automatically thresholded and filtered according to the preset algorithm to discriminate nonspecific staining using NIH Image J v 1.47 (Bethesda, MD). To control for variable Aβ parenchymal plaque load across APOE genotypes the Iba1 load and the CD68 load was divided by the load of Th-S positive Aβ parenchymal plaques determined on the same microphotograph as described above.
5
Hypoxia Promotes Cell Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single spheroids were placed on 8 µm pore inserts (Falcon) in serum-free medium for 4 h in 24-well companion plates (Falcon) and treated as indicated. Once adherent, HG medium with 4% FBS as chemoattractant was added in the bottom of each well and spheroids were either placed in normoxic (21% O2) or hypoxic (1-2% O2) conditions for 6 h. Spheroids were then gently removed washed from the top layer of the insert with PBS. Each insert was fixed in 100% ice-cold methanol for 15 min at -20 °C, washed 2x with PBS for 5 min and then mounted onto glass slides with Gold Antifade Mounting media with DAPI (ThermoFisher) and covered with 2 mm glass coverslips. Cells were imaged and counted using a Nikon 80i fluorescent microscope for each spheroid that was placed in the insert.
6
Golgi Apparatus Labeling in Yeast Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Golgi apparatus labeling, the pmr1/pmr1 GDT1-GFP/gdt1 cells were grown to log-phase in SD-URA, harvested, washed twice with PBS, and suspended in 200 μl PBS supplemented with 4 μl beta-mercaptoethanol and 15 U lyticase enzyme (Sigma). Cells was incubated at 37 °C for 40 min to partially digest the cell wall, and were washed once with PBS and suspended in 200 μl PBS before they were mixed with 2 μl Golgi-Tracker Red dye (33.3 mg ml− 1) (Beyotime Institute of Biochemistry, China) and incubated at 4 °C for 30 min. Cells were then washed twice and incubated in 1 ml SD-URA at 30 °C for 30 min, before they were visualized by the Nikon 80i fluorescent microscope.
7
Immunohistochemical Visualization of Microglia and Vagal Afferents
8
Immunohistochemistry of Tree Shrew Brains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tree shrew brains were embedded in paraffin and sectioned at a thickness of 5 μm on a microtome (Leica, RM2245). Slides were treated at 60 °C for 30–60 min, then deparaffinized in xylene and a gradient concentration of ethanol. Deparaffinized slides were rinsed with water and antigen retrieved in boiling citric acid buffer (PH = 6.0) for 24 min, which was boiled by microwave. After natural cooling, slides were washed sufficiently with water, immersed in 3 % H2O2 for 10 min, and permeabilized with 0.5 % TritonX-100 at RT for 10 min. Blocked in 5 % BSA at RT for 30 min, then incubated with diluted primary antibody (Rabbit anti HSV1, Cat. RR1190P, American Qualex Antibodies) at 4 °C, incubated overnight and subsequently diluted secondary antibody added (anti-rabbit IgG HRP-labeled Antibody, Cat #7074S, Cell Signaling Technology) at room temperature for 60 min. After development with DAB (Cat. DAB-0031) for 3 min, slides were washed in water and did counterstain with hematoxylin. Slides were dehydrated in a gradient concentration of ethanol and dehydrated with xylene, for 3–5 min each. Finally, slides were mounted with neutral balsam and the signal detected with a Nikon, 80i fluorescent microscope.
9
Cell Viability Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Aliquots for cell counts were taken at the 54 hour time point (described above). In the initial experiments, aliquots of the cells were stained with trypan blue, which only stains dead cells. Live and dead cells were then counted manually under a standard light microscope. Most experiments were done by staining aliquots of the cells with calcein AM (2 μM) and ethidium homodimer-1 (4 μM) from Life Technologies for 30 minutes at 37°C. Photographs were taken with a Nikon 80i Fluorescent Microscope (Tokyo, Japan). calcein AM stains live cells green under fluorescent lighting. ethidium homodimer-1 is impermeable to live cells and fluorescently stains dead cells red. The ITCN function in Image J from NIH was used to automatically count large numbers (1000+) of live and dead cells. The results were nearly identical using either method to count cells (data not shown).
10
Immunofluorescence Microscopy Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells grown on 8 well chamber slides (Nalge Nunc International) were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.2% Triton X-100, and blocked with Image-iT FX signal enhancer (Invitrogen) for 20 min. Cells were then incubated with primary antibodies against the specific proteins and subsequently stained with Alexa 488 or 594 conjugated secondary antibodies. Cells were mounted in mounting medium containing DAPI. Images were acquired using a Nikon 80i fluorescent microscope equipped with a Metamorph digital imaging system.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!