Automated Methods for the Analysis of Skeletal Muscle Fiber Size and Metabolic Type - International Review of Cell and Molecular Biology - page 288

muscle CSAs by Kalliainen et al. (2002) which utilized SigmaScan Image
software (SPSS Science, Chicago, IL) and the analysis of different muscle
types from monkeys by Singh et al. (2002) which utilized Image-Pro soft-
ware (Media Cybernetics, Silver springs, MD). Notably, MetaMorph soft-
ware is yet another commercially available program that was used
prominently by muscle researchers; in this initial mode, which required trac-
ing of the fiber outlines, MetaMorph was utilized by researchers including
Rafuse et al. (2000) who evaluated CSAs of muscle fibers in neural cell adhe-
sion molecule (NCAM) null mice, Bodine et al. (2001) and Lai et al. (2004)
who evaluated CSAs of fibers in rat and mouse muscle hypertrophy models,
and Hunter and Kandarian (2004) who evaluated CSAs in a mouse
atrophy model.
5.2.1 Semiautomated calculation of CSAs
Despite significant time requirements, semiautomated measurement of the
CSAs of fibers (in which a digitizing pen or computer mouse are used to
trace the fiber outlines in digital micrographs) is still extensively used in skel-
etal muscle research. This overall method of CSA quantification is
“semiautomated,” as it requires the investigator to manually interact with
the computer to trace the fiber outlines. For this purpose, perhaps the most
popular software that is currently used is Image J. Recent examples in which
Image J has been utilized for CSAmeasurements include Carroll et al. (2011)
who calculated CSAs of ATPase-stained fibers in soleus muscle of LG/J and
SM/J strains of mice, Wust et al. (2012) who evaluated the plantaris muscle
of rats with right ventricular heart failure, Taivassalo et al. (2012) who eval-
uated muscle fibers in skeletal muscle of patients with mitochondrial myop-
athy, and Hao et al. (2011) who studied muscle recovery from hind-limb
suspension in old rats.
Image-Pro software fromMedia Cybernetics (Bethesda, MD) is also fre-
quently used for semiautomated calculation of CSA. Examples include Rezk
et al. (2012) who analyzed the CSAs of H&E-stained diaphragm muscle
fibers and showed that angiotensin II infusion for 7 days significantly
reduced fiber size, McKiernan et al. (2012) who showed that calorie restric-
tion significantly increased Type I fiber CSAs but had no effect on Type II
fibers in vastus lateralis sections from rhesus monkeys, Zhao et al. (2011)
who evaluated CSAs in rabbit ischemic limb model, and Reed et al.
(2011) who evaluated CSAs of fibers in a rat muscle atrophy model.
The widespread use of semiautomated methods for measuring CSA
when several automated software for CSA measurements are available