LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 2 µM calcein AM and 4 µM EthD-1. Incubate the cells for 30–45 minutes at room temperature

Protocol tips
Add 2 µM calcein AM and 4 µM EthD-1. Incubate the cells for 30–45 minutes at room temperature

Publication protocol

Six culture substrates of Ti-Ap, H2O2–Ti-Ap, 300 °C-Ti-Ap, or Ti were placed on each well of a 24-well TCPS plate. Among these substrates, three were used for measuring cell attachment areas while the remaining three substrates were used for the evaluation of osteogenic differentiation. Rat MSCs were seeded on each cell culture substrate at a density of 4 × 104 cells ml−1 (1 ml well−1). For measuring the attachment areas of live cells, cells were prestained using a LIVE/DEAD viability/cytotoxicity assay kit. The cells attached to the surface of each culture substrate were observed 2 h after seeding under an objective fluorescence microscope (Olympus, Model BX51). Live cells stained with calcein-AM appeared green owing to intracellular esterase activity, while dead cells stained with EthD-1 appeared red owing to plasma membrane disintegrity. The attachment area of live cells on each cell culture substrate was measured using Image-Pro® PLUS software (Media Cybernetics, Inc., Version 7.0). Cell attachment areas between 350 and 3500 μm2 were used for analysis. Features smaller than 350 μm2 were regarded as dust particles and discarded. Areas larger than 3500 μm2 were regarded as multiply connected cells and discarded too.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Live / Dead assay mammalian cells - rat MSC using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Live / Dead assay mammalian cells - rat MSC using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.