| Strengths . |
|---|
| • Direct access to the sarcomere system |
| • Separation of cellular subsystems (e.g., sarcomeres versus sarcolemma) |
| • Ability to use fluorescent probes and other analytic tools |
| • Convenience of controllably performing different standardized experiments (e.g., isometric/isotonic contractions) |
| • Ability to perform protein exchange experiments that preserve overall functionality (e.g., troponin; Babu et al., 1988; Brenner et al., 1999; Gulati and Babu, 1989); and to probe time-resolve sarcomere dynamics by photolysis of caged compounds (ATP [Goldman et al., 1982, 1984], inorganic phosphate [Araujo and Walker, 1996; Dantzig et al., 1992; Millar and Homsher, 1990; Tesi et al., 2000], and Ca2+ chelators [Luo et al., 2002; Wahr et al., 1998]) |
| • Simpler handling and storage logistics (samples can be thawed and analyzed after prior freezing) |
| Weaknesses |
| • Challenge of reproducing the native physiological environment |
| • Variations in results between laboratories |
| • Instability and sensitivity to temperature |
| • Challenges of [Ca2+] calibration |
| • Structural changes caused by skinning (e.g., altered sarcomere morphology, loss of cellular heterogeneity), impacting functional behavior |
| Strengths . |
|---|
| • Direct access to the sarcomere system |
| • Separation of cellular subsystems (e.g., sarcomeres versus sarcolemma) |
| • Ability to use fluorescent probes and other analytic tools |
| • Convenience of controllably performing different standardized experiments (e.g., isometric/isotonic contractions) |
| • Ability to perform protein exchange experiments that preserve overall functionality (e.g., troponin; Babu et al., 1988; Brenner et al., 1999; Gulati and Babu, 1989); and to probe time-resolve sarcomere dynamics by photolysis of caged compounds (ATP [Goldman et al., 1982, 1984], inorganic phosphate [Araujo and Walker, 1996; Dantzig et al., 1992; Millar and Homsher, 1990; Tesi et al., 2000], and Ca2+ chelators [Luo et al., 2002; Wahr et al., 1998]) |
| • Simpler handling and storage logistics (samples can be thawed and analyzed after prior freezing) |
| Weaknesses |
| • Challenge of reproducing the native physiological environment |
| • Variations in results between laboratories |
| • Instability and sensitivity to temperature |
| • Challenges of [Ca2+] calibration |
| • Structural changes caused by skinning (e.g., altered sarcomere morphology, loss of cellular heterogeneity), impacting functional behavior |