Fixed brain tissues were washed with tap water for overnight, dehydrated by gradient of ethyl alcohol from 70 to 100%, and cleaned with xylene. Tissues were embedded with paraffin using embedding center (Leica, Wetzlar, Germany), sectioned into 4 μm coronal section on a rotary microtome (Leica), placed on slide glass, and dried on a slide warmer (Thermo Fisher Scientific, Waltham, MA, USA). Sections were depaffinized with xylene, rehydrated by gradient of ethyl alcohol from 100 to 70%, and stained with Harris’ hematoxylin solution (Sigma-Aldrich) and eosin Y solution (Sigma Aldrich). Stained sections were dehydrated by gradient of ethyl alcohol from 70 to 100%, cleaned with xylene, and mounted with permount mounting solution (Thermo Fisher Scientific). They were observed and photographed with Olympus microscope (Olympus, Tokyo, Japan).
Embedding center
Manufactured by Leica
Sourced in Germany
The Embedding Center is a laboratory equipment designed for the preparation of tissue samples for microscopic analysis. It allows for the embedding of tissue samples in a support medium, typically paraffin, to facilitate sectioning and further processing. The core function of the Embedding Center is to provide a controlled and consistent environment for the embedding process.
Automatically generated - may contain errors
Lab products found in correlation
Permount mounting solution, by Thermo Fisher Scientific (4 mentions)
Harris hematoxylin solution, by Merck Group (4 mentions)
Rotary microtome, by Leica (3 mentions)
Eosin y, by Merck Group (3 mentions)
Slide warmer, by Thermo Fisher Scientific (2 mentions)
Eosin y solution, by Merck Group (1 mentions)
Eosin solution, by Merck Group (1 mentions)
Hematoxylin solution, by Merck Group (1 mentions)
Permount solution, by Thermo Fisher Scientific (1 mentions)
6 protocols using embedding center
1
Paraffin-Embedded Tissue Sectioning and Staining
2
Histological Preparation of Brain Tissues
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Brain tissues were fixed in formaldehyde 4% with PBS (0.1 M) and washed with water. Tissues were dehydrated by graded ethyl alcohol series from 70 to 100%, cleaned with xylene and fixed in paraffin using an embedding center (Leica, Westlar, Germany). Paraffin blocks were cut into 4 μm segments, deparaffinized with xylene and hydrated by graded ethyl alcohol series (from 100 to 70%). Segments were stained with Harris’ hematoxylin solution (Sigma-Aldrich, St. Louis, MO, USA) for 3 min and Eosin Y (Sigma-Aldrich) for 1 min. Segments were washed with water, dehydrated with graded ethyl alcohol series, mounted (Thermo Fisher Scientific, Waltham, MA, USA) and photographed using an Olympus microscope (Olympus, Tokyo, Japan).
3
Tissue Preparation for Histological Analysis
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Fixed brain tissues were washed with flowing tap water, dehydrated by series of graded ethyl alcohol from 70 to 100%, and cleaned with xylene. Brain tissues were embedded in paraffin tank using embedding center (Leica, Westlar, Germany) and hardened as tissue blocks. Tissues were cut into 4 μm sections and placed on glass slides in tissue bath (Leica). Sections were dried on slide warmer (Thermo Fisher Scientific, Waltham, MA, USA), deparaffinized with xylene, and rehydrated by series of graded ethyl alcohol from 100 to 70%. They were stained with hematoxylin solution (Sigma-Aldrich) and eosin solution (Sigma-Aldrich), subsequently. Stained tissues were washed with tap water and dehydrated with series of graded ethyl alcohol. They were mounted with permount mounting solution (Thermo Fisher Scientific) and photographed using Olympus microscope (Olympus, Tokyo, Japan).
4
Histological Analysis of Fixed Cerebral Cortex
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Fixed cerebral cortex tissues were washed with tap water, dehydrated with gradient ethyl
alcohol series from 70% to 100%, and cleaned with xylene. Tissues were embedded into
paraffin using Embedding center (Leica, Westlar, Germany) and made into paraffin tissue
blocks. Tissue blocks were sectioned 4 µm thick using a rotary microtome (Leica) and
sliced sections were placed on slide glass. Tissue slides were deparaffinized with xylene
and hydrated with ethyl alcohol series from 100% to 70%, and were kept in water. Tissue
slide were stained with Harris’ hematoxylin solution (Sigma-Aldrich) for 3 min, washed
with tap water for 10 min, reacted with Eosin Y (Sigma-Aldrich) for 1 min. After staining
process, tissue slides were dehydrated with gradient ethyl alcohol series, cleaned with
xylene, and sealed with a permount mounting solution (Fisher scientific, Fair Lawn, NJ,
USA). The morphological changes of the cerebral cortex were observed using Olympus
microscopy (Olympus, Tokyo, Japan) and microscopic images were taken.
alcohol series from 70% to 100%, and cleaned with xylene. Tissues were embedded into
paraffin using Embedding center (Leica, Westlar, Germany) and made into paraffin tissue
blocks. Tissue blocks were sectioned 4 µm thick using a rotary microtome (Leica) and
sliced sections were placed on slide glass. Tissue slides were deparaffinized with xylene
and hydrated with ethyl alcohol series from 100% to 70%, and were kept in water. Tissue
slide were stained with Harris’ hematoxylin solution (Sigma-Aldrich) for 3 min, washed
with tap water for 10 min, reacted with Eosin Y (Sigma-Aldrich) for 1 min. After staining
process, tissue slides were dehydrated with gradient ethyl alcohol series, cleaned with
xylene, and sealed with a permount mounting solution (Fisher scientific, Fair Lawn, NJ,
USA). The morphological changes of the cerebral cortex were observed using Olympus
microscopy (Olympus, Tokyo, Japan) and microscopic images were taken.
5
Histological Processing of Brain Tissue
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Brain tissues were fixed in 4% formaldehyde with 0.1 M phosphate buffered saline) and
washed with tap water. Tissues were dehydrated by series of graded ethyl alcohol from 70
to 100%), cleaned with xylene, embedded in paraffin using embedding center (Leica,
Westlar, Germany). Paraffin blocks were cut into 4 µm sections and
sections were placed on glass slides. Sections were deparrafinized with xylene and
hydrated by graded with ethyl alcohol series from 100 to 70%. Sections were stained with
Harris’ hematoxylin solution (Sigma-Aldrich, St. Louis, MO, U.S.A.) for 3 min and Eosin Y
(Sigma-Aldrich) for 1 min. Sections were washed with tap water, dehydrated with graded
ethyl alcohol series, mounted with permount mounting solution (Thermo Fisher Scientific,
Waltham, MA, U.S.A.), photographed using Olympus microscope (Olympus, Tokyo, Japan).
washed with tap water. Tissues were dehydrated by series of graded ethyl alcohol from 70
to 100%), cleaned with xylene, embedded in paraffin using embedding center (Leica,
Westlar, Germany). Paraffin blocks were cut into 4 µm sections and
sections were placed on glass slides. Sections were deparrafinized with xylene and
hydrated by graded with ethyl alcohol series from 100 to 70%. Sections were stained with
Harris’ hematoxylin solution (Sigma-Aldrich, St. Louis, MO, U.S.A.) for 3 min and Eosin Y
(Sigma-Aldrich) for 1 min. Sections were washed with tap water, dehydrated with graded
ethyl alcohol series, mounted with permount mounting solution (Thermo Fisher Scientific,
Waltham, MA, U.S.A.), photographed using Olympus microscope (Olympus, Tokyo, Japan).
6
Histological Analysis of Neuronal Damage
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Fixed tissues were washed with tap water, dehydrated with gradient ethanol series from 70 to 100%, cleaned with xylene, and embedded in paraffin using embedding center (Leica, Westlar, Germany). Paraffin blocks were sliced into 4 μm thickness using a rotary microtome (Leica) and paraffin ribbons were placed on slide glass. Tissues were deparaffinized with xylene, rehydrated with gradient ethanol series from 100 to 70%, and immersed in water. They were stained with Harris’ hematoxylin solution (Sigma) for 3 min, washed with tap water for 10 min, stained with eosin Y solution for 1 min, and dipped with water. They were dehydrated with gradient ethanol series from 70 to 100%, cleaned with xylene, and mounted with permount solution (Thermo Fisher Scientific, Waltham, MA, USA). Stained tissues were observed under Olympus microscope (Olympus, Tokyo, Japan) and microscopic images were taken. Total neurons and damaged neurons were counted in five random areas (500 × 500 μm) and percentage of damaged neurons was calculated by following formula: The number of damaged neurons / Total neurons × 100.
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