Bz x700 light microscope

Samples of each fraction were smeared on glass slides (Muto Pure Chemicals, 5116-20F, Tokyo, Japan) using a wedge method and stained with Papanicolaou solution¹⁹ . The images were obtained either with a BX50F light microscope (Olympus, Tokyo, Japan) using 10 × and 20 × objective lenses (for the 0.3K, 2K, 10K, and 160K fractions) or with a Keyence BZ-X700 light microscope (Keyence, Osaka, Japan) using 20 × and 40 × objective lenses (for the OFs).

After the application of LISW, the mice were deeply anesthetized with an intramuscular injection of 100 mg/kg ketamine and 10 mg/kg xylazine. The brain was removed after decapitation, fixed in 4% paraformaldehyde for 24 h, and then embedded in paraffin. Sagittal sections (4-μm thick) were cut from the embedded brain and the lung coronal tissues and stained with haematoxylin–eosin (HE). To investigate the cause of lethality in the Upper neck group, Bodian staining was performed on the mice brains of this group as well as those of intact mice for pathological analysis, and the sections were observed using the BZ-X700 light microscope (Keyence Corporation, Osaka, Japan) under 2 × , 10 × and 100 × magnification.