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2015 ; 259
(2
): 80-96
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Developing 3D SEM in a broad biological context
#MMPMID25623622
Kremer A
; Lippens S
; Bartunkova S
; Asselbergh B
; Blanpain C
; Fendrych M
; Goossens A
; Holt M
; Janssens S
; Krols M
; Larsimont JC
; Mc Guire C
; Nowack MK
; Saelens X
; Schertel A
; Schepens B
; Slezak M
; Timmerman V
; Theunis C
; VAN Brempt R
; Visser Y
; Guérin CJ
J Microsc
2015[Aug]; 259
(2
): 80-96
PMID25623622
show ga
When electron microscopy (EM) was introduced in the 1930s it gave scientists
their first look into the nanoworld of cells. Over the last 80 years EM has
vastly increased our understanding of the complex cellular structures that
underlie the diverse functions that cells need to maintain life. One drawback
that has been difficult to overcome was the inherent lack of volume information,
mainly due to the limit on the thickness of sections that could be viewed in a
transmission electron microscope (TEM). For many years scientists struggled to
achieve three-dimensional (3D) EM using serial section reconstructions, TEM
tomography, and scanning EM (SEM) techniques such as freeze-fracture. Although
each technique yielded some special information, they required a significant
amount of time and specialist expertise to obtain even a very small 3D EM
dataset. Almost 20 years ago scientists began to exploit SEMs to image blocks of
embedded tissues and perform serial sectioning of these tissues inside the SEM
chamber. Using first focused ion beams (FIB) and subsequently robotic
ultramicrotomes (serial block-face, SBF-SEM) microscopists were able to collect
large volumes of 3D EM information at resolutions that could address many
important biological questions, and do so in an efficient manner. We present here
some examples of 3D EM taken from the many diverse specimens that have been
imaged in our core facility. We propose that the next major step forward will be
to efficiently correlate functional information obtained using light microscopy
(LM) with 3D EM datasets to more completely investigate the important links
between cell structures and their functions.
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