Live/Dead cell Staining Kit II

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 100 µl of the Calcein-AM/EthD-III to each well. Incubate the samples at room temperature for 30–45 minutes

Protocol tips
Add 100 µl of the Calcein-AM/EthD-III to each well. Incubate the samples at room temperature for 30–45 minutes

Publication protocol

To label cells by fluorescent dyes, 1 × 106 cells in the second passage of in vitro culture were detached by mild proteolysis (5 min, 37 °C, Accutase®), washed twice with PBS and incubated with PKH26 (red label) or PKH67 (green label) as described [55] using standard reagent kits (Sigma-Aldrich, Taufkirchen, Germany). To confirm high viability of MSCs in the attachment assays, cells were loaded with Calcein-AM and Ethidium-homodimer (Live/Dead cell Staining Kit II, PromoCell, Heidelberg, Germany). Then attachment of cells was explored with the labeled MSCs as described above. Live cells emitting green fluorescence at 515 nm and dead cells emitting red fluorescence at 620 nm were recorded by a fluorescence microscope

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