Automated Methods for the Analysis of Skeletal Muscle Fiber Size and Metabolic Type - International Review of Cell and Molecular Biology - page 286

measurements. In 1982, Round et al. described a flexible microprocessor
system for planimetry of muscle fiber CSAs ( Round et al., 1982 ); fiber out-
lines were drawn using tablet and drawing pen. While the computer capacity
was very small, it was a great advancement compared with previous CSA-
measuring techniques, and this method is somewhat similar to currently
widely used CSA measurement method using Image J software (see below).
Similar, semiautomated image analyzers were later used by other researchers,
including a Leitz ASM by Shorey and Cleland (1983) for measurements of
CSAs of fibers in rat muscle, and a Zeiss MOP Digital Image Analyzer by
Gollnick for histochemical slides of overloaded chicken muscle ( Gollnick
et al., 1983 ). Relatedly, Thomas et al. (1987) used a surface-imaging digitizer
interfaced with a microcomputer to measure CSAs of hamster muscle fibers
on Polaroid photomicrographs, and samples from human tibialis anterior
muscle were analyzed utilizing projection of images from a binocular micro-
scope to a digitized tablet interfaced to an ABC 806 computer ( Henriksson-
Larsen, 1985 ). Notably, measurements from human vastus lateralis muscle
were published in 1987, obtained utilizing an Interactive Image Analysis
System ( Poggi et al., 1987 ), and in 1993, for researchers utilizing Autosketch
2.0 ( Donnelly et al., 1993 ).
In 1982-1993, muscle biologists often used histochemically stained mus-
cle cross sections and numerous different systems for digitized planimetry to
measure CSAs of muscle fibers. Examples include the calculation of CSAs of
ATPase-stained muscle fibers from young and old rats using a Hewlett-
Packard 85 computer and HP9111A Graphics package ( Holloszy et al.,
1991 ), and the measurement of CSAs using gray-scale photographs of mus-
cle sections imaged frommuscle sections stained via periodic acid-Schiff and
digitized with a microcomputer ( Larsson et al., 1991 ).
5.1.3 Utilizing fluorescence microscopy in combination with planimetry
Starting in the 1990s, digitized planimetry of muscle was further advanced by
labeling and imaging techniques featuring fluorescence microscopy. In
1994, Stauber et al. used ConA-fluorescein lectin to label rat soleus muscle
sections, as the fluorophore-conjugated lectin, which binds to the plasma
membrane, produces sharp outlines of the muscle fibers ( Stauber et al.,
1994 ). The researchers prepared photomicrographs from the muscle sections
utilizing a standard camera and 35-mm slide film; images from the color
slides were then captured with a video camera/“frame-grabber” setup inter-
faced to a computer and the fiber outlines traced with a digitizing pen. This
method was further developed by Mozdziak et al. (1996) by combining