Potassium permanganate is an excellent alternative to osmium tetroxide in freeze-substitution.
Histochem Cell Biol
; 157(4): 481-489, 2022 Apr.
Article
em En
| MEDLINE
| ID: mdl-34984524
ABSTRACT
High-pressure freezing followed by freeze-substitution is a valuable method for ultrastructural analyses of resin-embedded biological samples. The visualization of lipid membranes is one of the most critical aspects of any ultrastructural study and can be especially challenging in high-pressure frozen specimens. Historically, osmium tetroxide has been the preferred fixative and staining agent for lipid-containing structures in freeze-substitution solutions. However, osmium tetroxide is not only a rare and expensive material, but also volatile and toxic. Here, we introduce the use of a combination of potassium permanganate, uranyl acetate, and water in acetone as complementing reagents during the freeze-substitution process. This mix imparts an intense en bloc stain to cellular ultrastructure and membranes, which makes poststaining superfluous and is well suited for block-face imaging. Thus, potassium permanganate can effectively replace osmium tetroxide in the freeze-substitution solution without sacrificing the quality of ultrastructural preservation.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Tetróxido de Ósmio
/
Permanganato de Potássio
Idioma:
En
Revista:
Histochem Cell Biol
Assunto da revista:
CITOLOGIA
/
HISTOCITOQUIMICA
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Alemanha