The disector is the only objective method for quantifying particles
of
variable size in a given volume. With
this method, cell organelles are identified on adjacent sections, but only
those present in one section are
counted. When counting extremely rare structures in transmission electron
microscope sections (physical
disector), the usual procedure of counting on electron micrographs is limited
for economic reasons (e.g.
micrographs highly outnumbering the investigated structures). Hence, to
apply
this unbiased stereological
method, a modification of the physical disector concerning 3 aspects has
been
developed. (1) The prerequisite
of screening large corresponding tissue areas (here
∼65000 μm2) was fulfilled by examining tissue areas
along the edges of ultrathin sections. (2) The size of the counting frame
was determined by measuring the
lengths of the section margins (minus a guard area) by means of a Morphomat.
This value was multiplied
by the width of the investigated tissue zone, corresponding to the diameter
of
the electron microscope
viewing screen. (3) Disector counting was carried out simultaneously on
both
sections (bidirectional disector)
to improve efficiency. In the present study tiny synaptic bodies (SBs)
were
quantitated by disector in a rat
pineal gland, yielding ∼30 SBs/1000 μm3. By contrast,
single section profile counts of SBs amounted to
90 SBs/20000 μm2. Since the presently described adaptation
of
the disector is time-consuming, it is proposed
to determine a proportion factor allowing to estimate number of structures
per
volume based on single section profile counts. This would decrease the
evaluation time by more than 50%.