Automated Methods for the Analysis of Skeletal Muscle Fiber Size and Metabolic Type - International Review of Cell and Molecular Biology - page 291

knockout ( Sheard et al., 2010 ), and WinROOF image-analysis software
(Mitani Corp., Fukui, Japan) which was used to evaluate the effects of ste-
roids and exercise in rat muscle ( Uchikawa et al., 2008 ). Finally, CyteSeer ®
(Vala Sciences Inc., San Diego—discussed in greater detail in Section 6.1 )
was recently used to evaluate changes in muscle associated with knockout
of CuZn-superoxide dismutase, an enzyme that possesses antioxidant activ-
ity ( Larkin et al., 2011 ).
6. APPLICATION OF HIGH-CONTENT ANALYSIS
METHODS TO ANALYSIS OF SKELETAL MUSCLE
A concept emerging from the development of digital microscopy is
“High-Content Analysis” (HCA), which refers to the rich treasure trove
of information represented by images of cells or tissue ( Jain and Heutink,
2010; Kummel et al., 2012 ). HCA microscopes photograph the cells or tis-
sue in multiple optical channels, including, for example, blue fluorescence
(often used in conjunction with DAPI-stained nuclei), green fluorescence
channel (for GFP and FITC-related fluorophores), red fluorescence channel
(for red fluorescent proteins and red fluorophores), and even far-red and
near-infrared channels ( Fig. 7.3 ). Digital pathology workstations (slide
Figure 7.3 The HCA analysis approach conceptually applied to analysis of skeletal mus-
cle. Muscle fiber outlines identified in one optical channel can be used to quantify bio-
markers labeled in additional optical channels, potentially including both fluorescence
and bright-field microscopy modes.