Journal of neuroscience methods · 2025
Optimized methyl green-pyronin Y staining for layer visualization in frozen mouse cerebellum.
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Kim Hyeju, Um Soo-Jong
Аннотация
BACKGROUND: Conventional histological stains, such as hematoxylin and eosin (H&E) or toluidine blue O (TBO), have a limited ability to clearly delineate the layered architecture of the cerebellar cortex.
NEW METHOD: We applied methyl green-pyronin Y (MGP) staining, which is traditionally used for nucleic acid differentiation, to frozen mouse cerebellar sections to enhance visualization of cortical layers and neuronal subtypes.
RESULTS: MGP staining yielded strong contrast between cell types: Purkinje cells stained distinctly pink, while granule cells appeared green. This enabled clear identification of cerebellar lamination and neuronal distribution.
COMPARISON WITH EXISTING METHODS: In H&E or TBO staining, Purkinje and granule cells are colored similarly, which obscures layer boundaries. Although immunohistochemistry is commonly used to distinguish these cell types, MGP staining provides a rapid, color-based distinction without the need for antibodies or fluorescence.
CONCLUSIONS: MGP staining provides a fast and cost-effective alternative for analyzing cerebellar tissue, enabling clear visualization of cortical layering and facilitating the morphological screening of cerebellar abnormalities.
NEW METHOD: We applied methyl green-pyronin Y (MGP) staining, which is traditionally used for nucleic acid differentiation, to frozen mouse cerebellar sections to enhance visualization of cortical layers and neuronal subtypes.
RESULTS: MGP staining yielded strong contrast between cell types: Purkinje cells stained distinctly pink, while granule cells appeared green. This enabled clear identification of cerebellar lamination and neuronal distribution.
COMPARISON WITH EXISTING METHODS: In H&E or TBO staining, Purkinje and granule cells are colored similarly, which obscures layer boundaries. Although immunohistochemistry is commonly used to distinguish these cell types, MGP staining provides a rapid, color-based distinction without the need for antibodies or fluorescence.
CONCLUSIONS: MGP staining provides a fast and cost-effective alternative for analyzing cerebellar tissue, enabling clear visualization of cortical layering and facilitating the morphological screening of cerebellar abnormalities.