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Fifty μm thick sequential sections were cut on a saw microtome (SP1600, Leica, Germany).
All biopsy specimens for histological examination were fixed in 10% formalin, embedded in paraffin wax on the oriented edge, and cut into 5 μm thick sequential sections.
For single immunohistochemical analysis, formalin fixed (4%, 24 hours) paraffin embedded 5 μm thick sequential sections were mounted on coated glass slides (Menzel Gläser Superfrost PLUS, Thermo Scientific, Braunschweig Germany), dried over night at 37°C and deparaffinised.
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Formalin-fixed, paraffin-embedded specimens were cut into 4 μm-thick sequential sections.
As described previously [ 11- 14], formalin-fixed, paraffin-embedded specimens were cut into 4 μm-thick sequential sections.
For this study, PEM films, only a few hundred nanometers thick, were assembled by sequential adsorption of poly acrylic acid) and poly allylamine hydrochloride).
For the hind limbs, the ROIs were drawn around sequential 1-voxel thick transverse slices using the blended SPECT/CT to define limb boundaries from the proximal femur to the distal tibia/fibula, and the activity from all the slices were summed for each limb.
Scanning was done in sequential 3 mm thick layers (for aorta starting scanning 3 cm proximal to the bifurcation of the aorta).
Sequential 10 μm thick coronal sections were made within 0.5 mm of the Bregma line for sections including striatum and between Bregma −1.5 and −2.0 for sections including hippocampus [ 21].
Sequential 10-μm thick coronal brain slices were made within 0.5 mm of the Bregma line for striatal sections and between Bregma −5.5 and −6.25 for brain stem sections (Franklin and Paxinos, 1997).
The acquisition parameters were: TR/TE = 3400/12 msec, TI1/TI2 = 700/1900 msec, FOV = 256 mm, 24 sequential 4-mm thick slices with a 25% gap between the adjacent slices, partial Fourier factor = 6/8, bandwidth = 2368 Hz/pix, and imaging matrix = 64 × 64.
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