Biomedical Engineering Reference
In-Depth Information
To avoid mistakes, it is necessary using stereomicroscope
to place the slot grid into the holder of EM and introduce it
into EM and then to fi nd the very fi rst serial EM sections using
the traces (usually letters) of the coordinated grid fi lled with
the resin. These are visible mostly on the fi rst two sections. If
the central position of the cell of interest within the pyramid is
used as a marker, identify the central cell within the sections. It
is important to take consecutive photographs (or grab the
images with a computer using a video camera) of the serial sec-
tions beginning from the very fi rst section since the organelle
of interest (just observed under the LSCM) appears and until
it is no longer seen. Having images from FL is useful to iden-
tify the cell of interest. If EM and FM images are refl ective, it
is necessary to change the refl ection position of FM images.
8. The example of CVLEM is shown in Fig.
4
.
Fig. 4
Identifi cation of the cell in prophase with the help of universal holder. Consecutive images of the same
cell (
black arrows
) under different magnifi cations. (
a
) Photo of the cell of interest after its embedding into Epon
under the light microscope. (
b
) The same cells at low magnifi cation in the EM section. (
c
) The holder embedded
into the Epon. (
d
) Ultrathin section of the cell of interest (
black arrow
) at higher magnifi cation. (
d
) The higher
magnifi cation of the area shown in
black box
in plate (
c
). (
e
) The higher magnifi cation of the area shown in
black box
in plate (
d
). (
g
) The higher magnifi cation of the area shown in
black box
in plate (
f
). (
h
) The higher
magnifi cation of the area shown in
black box
in plate (
g
). Nuclear pores (
red arrows
in (
h
)) are shown in the
prophase cells. Bar: 260 nm
