Astrogliosis was evaluated using glial fibrillary activating protein (GFAP; 1:1000; Sigma) immunohistochemistry. Mature neuron survival was evaluated using NeuN (1:500; Abcam) immunohistochemistry. A Nikon Eclipse Ti microscope with attached to the charge coupled device (CCD) camera and Nikon NIS Element BR software was used to image stained sections at 20× magnification. Imaged brain sections were analyzed for stain intensity via positive pixel density quantification using Photoshop software. Stroke‐affected region was morphologically identified and corresponds to the core and peri‐infarct, a 500‐µm boundary extending from the edge of the infarct core, medial and lateral to the infarct within the cortex.
Nis element br
Manufactured by Nikon
Sourced in Japan
NIS Element BR is a microscope imaging software developed by Nikon. It provides a comprehensive platform for acquiring, processing, analyzing, and managing microscope image data. The software offers a range of tools and functionalities to support various microscopy techniques and research applications.
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Lab products found in correlation
Neuronal nuclei (neun), by Abcam (2 mentions)
Eclipse ni u, by Nikon (2 mentions)
Sheep anti th antibody, by Merck Group (2 mentions)
Eclipse ti microscope, by Nikon (1 mentions)
Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions)
Plan fluor, by Nikon (1 mentions)
Ds ri2, by Nikon (1 mentions)
Plan fluor 100x, by Nikon (1 mentions)
Planfluor4, by Nikon (1 mentions)
Plan fluor 60, by Nikon (1 mentions)
Cryostat, by Leica (1 mentions)
Eclipse 50i, by Nikon (1 mentions)
Fda pi, by Merck Group (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Eclipse e400 microscope, by Nikon (1 mentions)
Alexa fluor conjugated goat anti rabbit or goat anti mouse antibody, by Thermo Fisher Scientific (1 mentions)
Anti c5b 9, by Abcam (1 mentions)
Fitc conjugated goat anti mouse igg h l, by LGC (1 mentions)
Ds qi2 camera, by Nikon (1 mentions)
Cage incubator, by Okolab (1 mentions)
12 protocols using nis element br
1
Evaluating Astrogliosis and Neuron Survival
2
Light Microscopy Measurements of Moss Cell Areas
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An upright light microscope (Nikon Eclipse Ni-U) was used in bright field and interference contrast mode. The instrument was equipped with the objectives Plan Fluor 4× (NA 0.13), Plan Apo 10× (NA 0.45), Plan Apo 20× (NA 0.75), Plan Fluor 40× (NA 0.75), Plan Fluor 60× oil immersion (NA 0.50–1.25), Plan Fluor 100x oil immersion (NA 1.30) and an attached camera (Nikon DS-Ri2). For picture processing, the software NIS-Element BR (Nikon), including an “extended focus” tool, was used. The calibrated measurements were directly exported to Excel (Microsoft Office 365).
The cell areas of P. patens, H. cupressiforme, P. schreberi, and P. purum were calculated using the formula of a rectangle. For P. affine, we used the area of a hexagon [36 ] as it fitted best to the cell shape of this species (Figure 4 C).
The cell areas of P. patens, H. cupressiforme, P. schreberi, and P. purum were calculated using the formula of a rectangle. For P. affine, we used the area of a hexagon [36 ] as it fitted best to the cell shape of this species (
3
Astrogliosis and Neuron Survival in Stroke
4
Islet Viability Analysis by FDA/PI
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Viability of 10 islets from each condition was analyzed by fluorescein diacetate/propidium iodide staining (FDA/PI, Sigma) by three independent investigators. The ratio of green to red cells provided the percentage of islet viability. Images were obtained on a Nikon Eclipse 50i microscope with Nis-Element-BR software (Nikon, Amstelveen, Netherlands).
5
Quantifying Sympathetic Axonal Density
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Mouse embryo thoraxes were dissected from paraformaldehyde fixed embryos, dehydrated by methanol series (50–100%), and incubated overnight in 3%H2O2 in 20%DMSO in methanol solution to quench endogenous peroxidase activity. Tissues were then rehydrated, and incubated with sheep anti-TH antibody (1:400; Millipore) for 3 days at 4°C, followed by HRP-conjugated secondary antibody for 3 days at 4°C. Immunoreactive signal was visualized by DAB detection system. Tissues were dehydrated by methanol series and cleared with benzyl benzoate/benzyl alcohol (2:1). Sympathetic axonal density was quantified by binary threshold level selection of the TH+ area in the superior vena cavae and the sinus venosus area in wholemount digital images using NIS-Element BR software (Nikon).
6
Immunofluorescence Staining of C5b-9 in Cells
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Cells were seeded onto a microscopy cover glass in 24-well tissue culture plates at a density of 100,000 cells/well. After culturing overnight, the culture medium was removed and washed with PBS. The cells were fixed in 4% paraformaldehyde and blocked with 3% bovine serum albumin (BSA) in PBS. Rabbit polyclonal anti-C5b-9 (Abcam) was used as the primary antibody. Cells were incubated with a primary antibody and then incubated with Alexa Fluor-conjugated goat anti-rabbit or goat anti-mouse antibody (Invitrogen, Carlsbad, CA, USA). Nuclei were stained using 4,6-diamidino-2-phenylindole (DAPI). Cells were mounted with Vectashield® (Vector Laboratories Inc., Burlingame, CA, USA) and examined under an Eclipse E400 microscope (Nikon Instruments Inc., Melville, NY, USA). Images were captured using a Nikon Digital site Fi3 and analyzed using NIS element BR (Nikon Instruments Inc.).
7
Quantifying Sympathetic Axonal Density
8
Quantification of Adipocytes and CLS
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Epididymal adipose tissue was fixed in a 10% neutral formalin solution and embedded in paraffin. Tissues were cut to a thickness of 6 μm and stained with hematoxylin and eosin (H&E). By using light microscopy (Eclipse Ni-U, Nikon, Tokyo, Japan) and an image analysis program (NIS-Element BR, Basic Research software, Nikon), the stained sections were analyzed to quantify the number and diameter of adipocytes and the number of CLS.
9
Neurite Length Measurement Protocol
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After selecting the concentrations of NA and NOB, cells were treated with NA 50 µM, NOB 50 µM, and the co-exposure of NA and NOB for 48 h. A phase-contrast microscope equipped with NIS Element BR software from Nikon (Melville, NY, USA) was used for the measurement of neurite length.
10
Immunofluorescence Microscopy of Chitosan Microparticles
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For immunofluorescence microscopy, 10 mg of freeze-dried chitosan microparticles were blocked overnight in 200 μl of PBS/5% skim milk at 4°C and then incubated overnight at 4°C with a mouse anti-Cap PCV2-specific monoclonal antibody (isotype IgG2a, Jeno Biotech Inc.) diluted 1:100 in PBS/0.1% Tween-20 (PBST). After washing with PBST, the microparticles were incubated with FITC-conjugated goat anti-mouse IgG (H + L) (Kirkegaard & Perry Laboratories Inc.) for 1 h. After further washing, the microparticles were visualized under a Nikon Eclipse E400 fluorescence microscope interfaced to a PC running capture software (Nis-Element Br, Nikon).
Microparticles loaded with extracts of S. cerevisiae transfected with an empty plasmid (S. cerevisiae/pYES2) were subjected to the same treatment and used as a negative fluorescence control.
Microparticles loaded with extracts of S. cerevisiae transfected with an empty plasmid (S. cerevisiae/pYES2) were subjected to the same treatment and used as a negative fluorescence control.
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