Paraffin Embedding and Thin Sectioning of Microbial Colony Biofilms for Microscopic Analysis.
J Vis Exp
; (133)2018 03 23.
Article
in En
| MEDLINE
| ID: mdl-29630036
ABSTRACT
Sectioning via paraffin embedding is a broadly established technique in eukaryotic systems. Here we provide a method for the fixation, embedding, and sectioning of intact microbial colony biofilms using perfused paraffin wax. To adapt this method for use on colony biofilms, we developed techniques for maintaining each sample on its growth substrate and laminating it with an agar overlayer, and added lysine to the fixative solution. These optimizations improve sample retention and preservation of micromorphological features. Samples prepared in this manner are amenable to thin sectioning and imaging by light, fluorescence, and transmission electron microscopy. We have applied this technique to colony biofilms of Pseudomonas aeruginosa, Pseudomonas synxantha, Bacillus subtilis, and Vibrio cholerae. The high level of detail visible in samples generated by this method, combined with reporter strain engineering or the use of specific dyes, can provide exciting insights into the physiology and development of microbial communities.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Paraffin Embedding
/
Biofilms
/
Microscopy
/
Microtomy
Language:
En
Journal:
J Vis Exp
Year:
2018
Document type:
Article