Table 1. Summary of mitochondrial isolation methods (Macroscale).

A table highlighting the relative benefits and disadvantages of different ‘macroscale’ mitochondrial methods of mitochondrial isolation, incorporating either density gradient centrifugation (DGC), differential centrifugation (DC) or none. Techniques compared relative to the performance of DGC alone*: ++ = higher; + = slightly higher or similar; - = lower; 2D-PAGE = two-dimensional polyacrylamide gel electrophoresis; AP = affinity purification; CE = capillary electrophoresis; FAOS = fluorescence activated organelle sorting; FFE = free flow electrophoresis; FFF = field-flow fractionation; LC-MS = liquid chromatography mass spectrometry; SEM = Scanning Electron Microscopy; TEM = transmission electron microscopy.

Isolation Method	Prior Fractionation	Mitochondrial Yield *	Mitochondrial Purity *	Starting Material *	Expense *	Throughput *	Subcellular Spatial Precision	Serial Sampling	Confirmation	General Comments	References
AP	DC (or none)	++ (++)	+ (++)	- (-)	++ (++)	++ (++)	No	No	Western blotting, immunofluorescence, qPCR	High selectivity; sonication may impact mitochondria viability; not suitable for larger samples	Hornig-Do et al. (2009); Ahier et al. (2018) and Hubbard et al. (2019)
FAOS	DC	++	++	++	+	++	No	No	qPCR, NGS	High selectivity; shear damage and dyes may impact mitochondrial viability	Daniele et al. (2016)
FFE	DC (and DGC)	+ (-)	++ (++)	+ (+)	++ (++)	++ (++)	No	No	Western blotting, TEM	High mitochondrial viability; high selectivity	Zischka et al. (2006)
FFF	DC	++	-	+	++	++	No	No	Western blotting, SEM, 2D-PAGE, LC-MS	High mitochondrial viability; high selectivity; high working range	Kang et al. (2008) and Yang et al. (2015)
CE	DC	-	++	-	++	++	No	No	qPCR	High selectivity; predisposed to clogging	Poe et al. (2006) and Poe et al. (2010)