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Find answers to frequently asked questions on LifeAct actin visualization.

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Watch our movie LifeAct: Actin Marker for Live Cell Imaging (MV19).

Advantages of Using LifeAct for Actin Visualization



LifeAct specifically stains F-actin and perfectly co-localizes with existing markers.

 

Phalloidin

FP-Actin

LifeAct

Fixed Samples

++

n.a.

++
(Peptide)

Live Cell Imaging

-
(Toxin)

++

++

Influence on
Actin Dynamics

- -
(Toxin)

-
(High)

++
(None)

Signal-to-Noise
Ratio

++

+

++


In contrast to the current approaches, LifeAct® visualizes F-actin within fixed and living cells, without compromising cellular processes, such as the locomotion of dendritic cells or neuronal polarization.

 

LifeAct, a 17-amino acid peptide, is derived from a protein found in Saccharomyces cerevisiae. As a novel actin marker, LifeAct combines crucial characteristics that make it the best marker available today. To date, it is the shortest actin marker for living cells and has a low binding affinity to F-actin. Although it is high enough to label F-actin very specifically, LifeAct does not disturb actin kinetics. Because of this, actin’s functionality in cellular processes remains unrestricted. So, by using LifeAct for visualization of the actin cytoskeleton, researchers achieve the most authentic results.

LifeAct Plasmids can also be transiently transfected into various cell types. Moreover, they allow for the generation of stable cell lines, which results in long-term actin staining for different applications. Difficult-to-transfect cells, such as primary cells, are easily transduced with the LifeAct Adenoviral Vectors. LifeAct Lentiviral Vectors mediate efficient transduction, integration, and long-term expression into dividing and non-dividing cells, both in vitro and in vivo. A stable LifeAct-TagGFP2 expressing HT-1080 cell line allows for long-term actin visualization without interference with cytoskeletal dynamics.

To readily achieve high imaging performance, transfection or transduction of LifeAct can easily be done in ibidi’s μ-Slides and μ-Dishes. With the Torpedo transfection Reagents, ibidi provides the ideal reagents - cationic lipids - and protocols for cell transfection. Torpedo DNA was optimized specifically for the transfection of mammalian cells with plasmid DNA in ibidi's µ-Slides (e.g., μ-Slide VI 0.4). Torpedo siRNA enables successful gene silencing experiments when using ibidi's µ-Slides in live cell imaging.


Live cell imaging of actin dynamics in primary dendritic mouse cells, transfected with pLifeAct-TagGFP2