Table 1.

Summary of all the experimental variables tested, their results and recommendations

Objective	Experimental Variable	Results	Recommendation
Density-mediated cell separation	Ficoll-Paque layer vs. Pellet	Reduced myocardial debris	Straining of cardiac digests through 40-μm cell strainers
		70–80% loss of leukocytes in pellets
Cell fixation	1% PFA	Comparable with unfixed sample	Fixation using 1% PFA for 10 min at 4–8°C
	Ice-cold methanol/acetone	Decreased antibody binding
		Increased granularity due to protein denaturation
Intravascular cells	Coronary perfusion	Removal of 90–95% intravascular cells	Low-speed centrifugation at 50 g for 2 min after fine mincing of the tissue, and discard the supernatant containing intravascular cells
	Low-speed centrifugation post-tissue mincing	Removal of 90–95% intravascular cells
		Majority of intravascular cells are removed in the supernatant.