Imaging Internal Features of Whole, Unfixed Bacteria (Scanning, 2011)
Wet scanning-transmission electron microscopy (STEM) is a technique that allows
high-resolution transmission imaging of biological samples in a hydrated state,
with minimal sample preparation. However, it has barely been used for the study of
bacterial cells. In this study, we present an analysis of the advantages and disadvantages
of wet STEM compared with standard transmission electron microscopy (TEM). To investigate
the potential applications of wet STEM, we studied the growth of polyhydroxyalkanoate and
triacylglycerol carbon storage inclusions. These were easily visible inside cells, even
in the early stages of accumulation. Although TEM produces higher resolution images, wet
STEM is useful when preservation of the sample is important or when studying the relative
sizes of different features, since samples do not need to be sectioned. Furthermore, under
carefully selected conditions, it may be possible to maintain cell viability, enabling new
types of experiments to be carried out. To our knowledge, internal features of bacterial cells
have not been imaged previously by this technique.
(Right image: Fluorescence microscopy highlights polyhydroxyalkanoate inclusions
inside bacteria; optical image left, fluorescence image right)