Exosome diameter was determined by TEM and NanoSight particle tracking analysis. For TEM, undiluted exosomes were adsorbed onto formvar/carbon-coated copper grids, counterstained with uranyl acetate, and imaged at 50,000× and 150,000× magnification using a Hitachi 7650 analytical TEM paired with an 11-megapixel Erlangshen digital camera. For NanoSight size distribution analysis, exosomes were diluted 1:1,000 in molecular grade water (Corning, 46-000-CM) and analyzed using a NanoSight NS300 (NTA version 3.1, Malvern). For all samples, camera level = 12, frame rate = 25 FPS, acquisition time = 3 × 60 seconds, viscosity = water (~0.9 cP), detection threshold = 5, blur size = auto, max jump distance = auto.
7650 analytical tem
Manufactured by Hitachi
The 7650 Analytical TEM is a transmission electron microscope (TEM) designed for analytical applications. It provides high-resolution imaging and analytical capabilities for a variety of materials and samples. The core function of the 7650 Analytical TEM is to enable detailed examination and analysis of specimen structures at the nanoscale level.
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Lab products found in correlation
3 protocols using 7650 analytical tem
1
Exosome Size Characterization by TEM and NanoSight
2
Ultrastructural Analysis of Tendon Fibrils
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FDL tendons were isolated (n = 3 per diet or genotype) and fixed in Glutaraldehyde Sodium Cacodylate fixative. One-micron axial sections were cut and stained with Toluidine blue. One-micron sections were then trimmed to 70 nm and stained with uranyl acetate and lead citrate. Sections were placed on grids for imaging on a Hitachi 7650 Analytical TEM. Eight to twelve non-overlapping images were taken from mid-substance of each tendon at 15,000x and 40,000x magnification. For measurement of fibril diameter, a region of interest (ROI) of was determined within each image so that a minimum of 80 fibrils could be measured. Diameters were measured along the y-axis. Collagen Fibril density was measured in a 2340 × 1860 pixel area. Collagen fibril density and diameter measurements were made in ImageJ.
3
Ultrastructural Analysis of Tendon Fibrils
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FDL tendons were isolated (N = 4 for WT; N = 3 for ScxLinDTR,10weeks) and fixed in Glutaraldehyde Sodium Cacodylate fixative. One-micron axial sections were cut and stained with Toluidine blue. One-micron sections were then trimmed to 70 nm and stained with uranyl acetate and lead citrate. Sections were placed on grids for imaging on a Hitachi 7650 Analytical TEM. Three non-overlapping images were taken from mid-substance of each tendon at ×40,000 magnification. For measurement of fibril diameter, a region of interest (ROI) was determined within each image so that a minimum of 80 fibrils could be measured. Diameters were measured along the y-axis. The perimeter and the area of the collagen fibrils were quantified. The radii based on the calculated perimeter and area were quantified. The ratio of these two radii represent a measure of fibril roundness (fibril irregularity factor; FIF). An FIF different than one suggest that the fibril is not a perfect circle.
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