Mice were sacrificed by the intraperitoneal injection of sodium pentobarbital and exsanguination by aortic transection. After aortic transection, a thoracotomy was performed, the right bronchus was ligated, and the right lungs were removed and snap frozen. The tracheas were cannulated, and the left lungs were inflated and fixed with 4% formalin (G1101, Servicebio, China) at a pressure of 25 cmH2O for ≥15 min. After equilibration, the left lungs were removed and fixed in 4% formalin overnight. Subsequently, lung tissues were cut into 5-μm-thick sections on a Leica model 2165 rotary microtome (Leica, Nussloch, Germany) and stained with hematoxylin and eosin (H&E) as previously described45 (link),46 (link).
Model 2165 rotary microtome
Manufactured by Leica
Sourced in Germany
The Leica model 2165 rotary microtome is a laboratory instrument designed for the precise sectioning of samples for microscopic examination. It features a rotary mechanism that allows the user to obtain thin, uniform sections of materials such as biological tissues or other solid samples.
Automatically generated - may contain errors
9 protocols using model 2165 rotary microtome
1
Lung Tissue Fixation and Histology
2
Histological Analysis of Wound Tissue
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After 30 days, the wound tissues were harvested and then fixed in 10% buffered formaldehyde solution for 48 hours. Then, the product was decalcified in 10% neutral EDTA for approximately 4 weeks and finally embedded in paraffin. The paraffin blocks were sectioned into 5‐µm‐thick slices by a Leica model 2165 rotary microtome (Leica) and placed on glass slides. The deparaffinized tissues were stained with haematoxylin for 15 minutes, and then washed with distilled water. Afterwards, the tissues were immersed in 1% hydrochloric acid‐alcohol for 5 minutes and stained with eosin for another 5 minutes. After dehydration, the slices were sealed with neutral gum and observed by microscopy.
3
Lung Histological Examination Protocol
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After the BALF was collected, PBS was pumped into the right ventricle to clear the blood in the pulmonary vasculature. Then, the upper right lung lobe was removed and fixed in 10% neutral buffered formalin for 24 h; then, the specimens were dehydrated and embedded in paraffin. To carry out a histological examination, 5 μm sections of fixed embedded tissues were cut on a Leica model 2165 rotary microtome (Leica, Nussloch, Germany) and stained with hematoxylin and eosin. Histological analyses were performed by two independent pathologists blinded to the treatment groups.
4
Lung Tissue Fixation and Histological Analysis
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Mice were sacrificed by intraperitoneal injection of sodium pentobarbital. After aortic transection, a thoracotomy was performed, the right bronchus was ligated, and the right lungs were removed and snap frozen. The tracheas were cannulated, and the left lungs were inflated and fixed with 4% paraformaldehyde (G1101, Servicebio, China) at a pressure of 25 cm H2O for ≥15 min. After equilibration, the left lungs were removed and fixed in 4% formalin overnight. Subsequently, paraffin embedded lung tissues were cut into 5 μm-thick sections on a Leica model 2165 rotary microtome (Leica, Nussloch, Germany) and stained with H&E as previously described25 (link),26 (link).
5
Lung Tissue Histological Analysis
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After BALF was obtained, the upper right lung lobe was removed, fixed in 10% neutral buffered formalin for 24 h, and then specimens were dehydrated and embedded in paraffin. Five-micrometer sections of fixed embedded tissues were cut on a Leica model 2165 rotary microtome (Leica, Nussloch, Germany) and stained with Hematoxylin and Eosin. Histological analyses were performed by two independent pathologists blinded to the treatment groups.
6
Histological Examination of Lung Tissue
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Four hours after the last inoculation with saline or VP1, PBS was pumped into the right ventricle to clear the blood in the pulmonary vasculature. Then, the left lower lung lobe was excised, fixed in 4% paraformaldehyde for 24 h, embedded in paraffin, cut into 5 μm sections on a Leica model 2165 rotary microtome (Leica, Nussloch, Germany), and stained with H&E. For each animal, five fields were measured randomly from the H&E staining at a magnification of ×200.
7
Histological Analysis of Mouse Lung and Trachea
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Forty-eight hours after the last OVA challenge, mice were killed. The lung and trachea of mice were filled with 10% (v/v) neutral buffered formalin intratracheally and then were removed from the mice. For fixation, the neutral buffered formalin was also used. Specimens were dehydrated and embedded in paraffin. For histological examination, 4-μm sections of fixed embedded tissues were cut on a Leica model 2165 rotary microtome (Leica Microsystems Nussloch GmbH, Wetzlar, Germany), placed on glass slides, deparaffinized and stained sequentially with hematoxylin 2 and eosin-Y (Richard-Allan Scientific, Kalamazoo, MI, USA). Stained slides were analyzed using a light microscope (Axio Imager M1, Carl Zeiss) under identical conditions, including magnification ( × 20), gain, camera position, and background illumination.51 (link)
8
Histological Assessment of LPS-Induced Lung Injury
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At 48 hours after the instillation of LPS, mice were euthanized for histological assessment. The lung and trachea of mice were removed from the mice, and then for fixation, 10% (v/v) neutral buffered formalin was used. Specimens were dehydrated and embedded in paraffin. For histological examination, 4-μm sections of fixed embedded tissues were cut on a Leica model 2165 rotary microtome (Leica Microsystem Nussloch GmbH, Wetzlar, Germany), placed on glass slides, deparaffinized, and stained sequentially with H&E (Richard-Allan Scientific, Kalamazoo, MI, USA). Stained slides were analyzed with a light microscope (Axio Imager M1, Karl Zeiss, Goettingen, Germany) under identical conditions, including magnification (× 10), gain, camera position, and background illumination.
9
Histological Evaluation of Mouse Lungs
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Forty-eight hours after the HDM challenge, mice were euthanized. The lung and trachea of each mouse were filled with 10% (volume/volume) neutral buffered formalin intratracheally and were then removed. For fixation, neutral buffered formalin was also used. Specimens were dehydrated and embedded in paraffin. For histological examination, 4-μm sections of fixed and embedded tissues were cut on a Leica model 2165 rotary microtome (Leica Microsystems Nussloch GmbH, Wetzlar, Germany), placed on glass slides, deparaffinized, and stained sequentially with haematoxylin and eosin (Richard-Allan Scientific, Kalamazoo, MI, USA) or periodic acid-Schiff (PAS). Stained slides were analyzed using a light microscope (Axio Imager M1; Karl Zeiss, Goettingen, Germany) under identical conditions including magnification (× 20), gain, camera position and background illumination.18 (link)
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