The lung collected at 10 days after challenge was fixed with 10% normal buffered formalin (NBF) in PBS. After fixation NBF tissues were transferred to 70% alcohol and processed to paraffin using a Shandon Excelsior AS (Thermo Fisher Scientific). A standard program of dehydration was carried out in ethanol 70%, 80%, 90%, 95%, 100%, 100% 1 hour each, followed by xylenes twice for 30 min each. Finally, the tissue was soaked in paraffin for 30 min and repeated once, then held in the second paraffin bath for 1 hour. Left lung tissue sections (4 mice per group) were stained with hematoxylin and eosin (H&E) to assess histopathological changes as described (Ko et al. 2016 (link)). Images were acquired by Axiovert 100 (Zeiss, Oberkochen, Germany) at 100X magnification, and inflammation was blindly scored as previously described (Ko et al. 2016 (link)).
Shandon excelsior as
Manufactured by Thermo Fisher Scientific
The Shandon Excelsior AS is a tissue processor designed for the automated processing and dehydration of tissue samples. It features a compact design and is capable of processing up to 150 cassettes or 300 small samples simultaneously. The Excelsior AS automates the critical steps of tissue processing, ensuring consistent and reliable results.
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Lab products found in correlation
3 protocols using shandon excelsior as
1
Lung Histopathology Assessment Post-Challenge
2
Histopathological Assessment of Lung Samples
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The lungs collected at 5 day after challenge were fixed with 10% formalin in PBS. After fixation, the tissues were transferred to 70% alcohol and processed using a Shandon Excelsior AS (Thermo Fisher Scientific) and a standard program of dehydration. Finally, the tissue was soaked in paraffin for 2×30 min and then in the second paraffin bath for at least 1 h. Lung tissue sections were stained with hematoxylin and eosin (H&E) and periodic acid-schiff stain (PAS) to assess histopathological changes and mucin expression respectively as described previously [31 (link), 33 (link)]. Numerical assessment of histopathology scoring was based on a scale of 0–5 by blinded observers with the severity scoring system similar to the one as previously described [34 (link)]. Based on an ordinal scale, a score “<1” is represented within a normal or naive range whereas a maximum score of “5” is represented as extensive or severe histopathologic changes and infiltrates [34 (link)] (lymphocytes, polymorphonuclear cells, macrophages, eosinophils) in peribronchiolar, perivascular, interstitial, and alveolar spaces [31 (link), 33 (link)]. The mucin expression was detected with PAS-positive area.
3
Histopathological Analysis of Mouse Lungs
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