The number of Aβ42-positive cells was determined from sections of brain tissue from four animals in each of the treatment groups. Sections of paraffin-embedded tissue were immunostained with anti-Aβ42 antibodies under identical conditions. Five random images at 20X magnification from each animals’ cortical region were recorded under identical illumination, magnification and camera settings using a Nikon FXA microscope equipped with a Nikon CCD camera. Similarly, Aβ42-positive cells were visualized in the five random images at 20X magnification of the frozen brain cortical region using fluorescence settings of Nikon FXA microscope and Nikon Immunofluorescence camera. The total number of Aβ42-positive cells in each 20X viewing field was counted. Various statistical tools were used to test the significance of differences among the different treatment groups.
Fxa microscope
Manufactured by Nikon
Sourced in Japan
The Nikon-FXA microscope is a high-performance laboratory instrument designed for detailed observation and analysis. It features advanced optics and precision engineering to provide clear, high-resolution images. The core function of the Nikon-FXA microscope is to enable detailed examination and study of specimens across a wide range of scientific and research applications.
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7 protocols using fxa microscope
1
Quantifying Amyloid-β Positive Cells
2
SIV+ RNA Cell Quantification in Lymph Nodes
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Productively infected cells (SIV+ RNA cells) were assessed in LNs by in situ hybridization, as previously described.16 (link) Infected cells were detected and counted in the paracortical zone on a minimum of three sections using a Nikon-FXA microscope. A 35S-labeled RNA probe derived from the SIVmac nef gene was used. To enhance probe penetration into the tissue sections, the 35S-labeled RNA was subjected to mild alkaline hydrolysis to obtain a majority of fragments in the 150- to 200-nucleotide range. The number of positive cells was then divided by the surface of the entire LN section, and the results were expressed as the number of positive cells per 2-mm2 section. The mean count was calculated for three slides of the same LNs obtained in a blinded fashion by two different investigators.
3
Histological Analysis of IBA and 3,4-MDPU Effects
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For histological examination, basal segments (0.5 cm) were collected from the shoots treated with 2450 or 4900 μM IBA for 1 min and supplemented with 0, 0.1, 1 or 10 μM 3,4-MDPU for 24 h. Samples were collected after treatment for 5 days. Four samples were obtained for treatment, and the entire experiment was repeated twice. Histological analysis was performed following the protocol described by Ballester at al. (1999) (link). Briefly, explants were fixed in a mixture of formalin, glacial acetic acid and 50% ethanol [1:1:18 (v/v/v)], dehydrated through a graded n-butanol series and embedded in paraffin wax. Tissues were sectioned (10 μm) on a Reichert-Jung rotatory microtome. Sections were double-stained with safranin-fast green (Jensen 1962) . The stained sections were mounted with Euckit®, and images were captured with an Olympus DP71 digital camera fitted to a Nikon-FXA microscope.
4
Leaf Ultrastructure Preparation and Imaging
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Leaf samples were fixed at 4º C in a mixture of 2% glutaraldehyde and 2% paraformaldehyde in 0.05M sodium phosphate buffer (pH 6.8). After 48 h, samples were rinsed in the same buffer three times (20 min each), dehydrated in a graded aqueous ethanol series (30, 50, 70, 96, 100% 24h at 4ºC each step) and were progressively infiltrated with LR White resin (LRW; London Resin Company, England), according to the following schedule: 25, 50, 75% LRW/ethanol for at least 24 h each step, and two incubations in 100% LRW. Finally, samples were placed in plastic capsules containing 100% LRW and incubated for approximately 14 h at 60°C. Two-µm thick sections were cut using a Reichert-Jung ultra-microtome (Leica, Heidelberg, Germany).
Sections were stained with 0.05% Toluidine Blue (O'Brien and McCully, 1981) .
The stained sections (paraffin and resin) were mounted with Eukkit®, and photomicrographed with an Olympus DP71 digital camera (Japan) fitted to a Nikon-FXA microscope (Japan). 5 to 10 representative samples of each stage were analyzed.
Sections were stained with 0.05% Toluidine Blue (O'Brien and McCully, 1981) .
The stained sections (paraffin and resin) were mounted with Eukkit®, and photomicrographed with an Olympus DP71 digital camera (Japan) fitted to a Nikon-FXA microscope (Japan). 5 to 10 representative samples of each stage were analyzed.
5
Quantitative Ki-67 Expression Evaluation
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The evaluation of Ki-67 expression was conducted using a quantitative method. Immunoreactivity was considered when labeling occurred in the nucleus, regardless of the intensity [52 (link)]. Detailed observation of each preparation was conducted, choosing the tumor area with the highest nuclear positivity and staining homogeneity. Each selected area was analyzed using a 40× objective, and the fraction of positive nuclei was determined in terms of percentage, accounting for at least 1000 tumor cells in 8 to 10 fields. The same observer performed all counts using a Nikon FXA® microscope with a checkerboard eyepiece, considering cell morphology [52 (link)]. Subsequently, tumors were classified into two groups (low and high), using the mean Ki-67 positivity values as the cutoff.
6
Angiogenesis Regulation in 3D Fibrin Matrices
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Four independent hMVEC donors were cultured in normoxia until confluency, pooled together and seeded on top of 3D fibrin matrices. The hMVECs were transfected with specific si-RNA against the individual genes 4 h after seeding and 18 h later stimulated with VEGF-A/TNFα and transferred to hypoxia. Seven days after stimulation with VEGF-A/TNFα, two researchers evaluated the number of sprouts independently by eye and only the genes that were scored as more or less sprouts compared with the scrambled control by both researchers were selected for further investigation. In addition, invading cells and the formation of tubular structures of endothelial cells in the 3D fibrin matrix were analyzed by phase contrast microscopy. The total length of tube-like structures of triplicate wells was measured using 4 randomly chosen semi-dark field pictures/well using a Nikon FXA microscope equipped with a monochrome CCD camera (MX5). After threshold setting and skeletonization, the sprout formation was expressed as mm/cm2 using Optimas image analysis software (Adept Turnkey, Sydney, Australia).
7
Visualizing Mitotic Spindle Defects
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