Maintenance of Fluorescence During Paraffin Embedding of Fluorescent Protein-Labeled Specimens.

ABSTRACT: Paraffin embedding is widely used in microscopic imaging for preparing biological specimens. However, owing to significant fluorescence quenching during the embedding process, it is not compatible with fluorescent-labeling techniques, such as transgenic and viral labeling using green fluorescent protein (GFP). Here, we investigate the quenching mechanism and optimize the embedding process to improve the preservation of fluorescence intensity. The results show that dehydration is the main reason for fluorescence quenching during paraffin embedding, caused by the full denaturation of GFP molecules in ethyl alcohol. To evaluate fluorescent and morphological preservation, we modified the embedding process using tertiary butanol (TBA) instead of ethyl alcohol. Fluorescence intensity following T

SUBMITTER: Zhanmu O 

PROVIDER: S-EPMC6664058 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature