Cytoskeleton structure

In addition to acting as a cellular scaffold, the cytoskeleton has roles in organelle transport, cell division, motility, and signaling, making it central to both cell health and disease processes. Cytoskeleton stains routinely serve as fiducial markers in the fluorescence imaging of live and fixed cells for both orientation and colocalization. In addition, a range of dye conjugates and fluorescent proteins that react with cytoskeletal subclasses are commonly used in cytoskeleton research. Four distinct Invitrogen technologies are routinely used for imaging of actin, talin, or tubulin of the cytoskeleton.

CellLight reagents for the cytoskeleton

CellLight reagents are designed to label actin, tubulin, or talin in live cells, enabling researchers to follow cytoskeletal dynamics. CellLight proteins are compatible with other fluorescent probes for multiplex analysis in live cells, or after fixation for colocalization studies. These ready-to-use constructs express fluorescent fusion proteins targeted to the N-terminus of beta-actin (41.8 kDa), the N-terminus of beta-tubulin (49.8 kDa), or the C-terminus of talin (40 kDa). Introducing CellLight reagents involves a simple transfection step, and they work like cell stains with minimal toxicity or chemical disruption.

Phalloidin conjugates for actin staining

Tubulin Tracker dyes for live-cell microfilament staining

Taxol/paclitaxel and taxotere/docetaxel conjugates are used for endpoint assays of cytoskeletal behavior in live cells to provide intense staining of polymerized tubulin. This method is not generally used for dynamic cell imaging because it interferes with cellular functions, including the dynamics of tubilin instability and cell division. Tubulin Tracker Green reagent (paclitaxel conjugate) and Tubulin Tracker Deep Red reagent (docetaxel conjugate) are capable of passing through live-cell plasma membranes to specifically stain polymerized tubulin. Staining is lost during cell fixation.

Antibodies to actin and tubulin

Live cell staining of actin cytoskeleton

The Invitrogen CellMask Actin Tracking Stains allow for fluorescent staining of polymerized actin filaments (F-actin) in live cells. CellMask Actin Tracking Stains are cell permeable and provide specific, bright and uniform staining of the actin cytoskeleton. Following live cell staining, cells can be fixed for multiplexing applications in immunofluorescence (IF), immunocytochemistry (ICC), and immunohistochemistry (IHC) protocols. For flexibility in experimental designs, CellMask Actin Tracking Stains are offered in three colors, Green, Orange and Deep Red. Workflows are as simple as adding 1X staining solution to cells, incubating for 15-60 minutes, and finished by washing and imaging. CellMask Actin Tracking Stains cause no detectable cytotoxicity for up to 24 hours.

Selection guides

 
Readout
The easiest fluorescent staining method for actin; no calculations, no dilutions, no pipetting
 
Target
High-affinity staining of F-actin
Common filter set
FITC
TRITC
Ex/Em (nm)
495/518
555/565
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing
Yes
Yes
Live cells
No
No
Fixed cells
Yes
Yes
Fixable
Yes
Yes
Platform
Imaging
Imaging
Protocol Microscopy Microscopy
Format
6 x dropper bottles
6 x dropper bottles
Cat. No.
 
Readout
Expression of fluorescent fusion protein
Range
β-actin
β-tubulin 
Talin
Common filter set
FITC
FITC
FITC
Labels
GFP
GFP
GFP
Ex/Em (nm)
488/520
488/520
488/520
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing
Yes
Yes
Yes
Live cells
Yes
Yes
Yes
Fixed cells
No
No
No
Fixable
Yes
Yes
Yes
Platform
Imaging 
Imaging 
 Imaging
Format
1 mL
1 mL
1 mL
Cat. No.
 
Readout
 Expression of fluorescent fusion protein
Range
β-actin
β-tubulin 
Talin
Common filter set
TRITC
TRITC
TRITC
Labels
RFP
RFP
RFP
Ex/Em (nm)
555/584
555/584
555/584
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing
Yes
Yes
Yes
Live cells
Yes
Yes
Yes
Fixed cells
No
No
No
Fixable
Yes
Yes
Yes
Platform
Imaging 
Imaging 
 Imaging
Format
1 mL
1 mL
1 mL
Cat. No.
 
Readout
High-performance fluorescent label with stable signal, resistant to photobleaching 
Traditional fluorescent label 
Range
 F-actin
Common filter set
DAPI
DAPI
FITC
TRITC
TRITC
Labels
Alexa Fluor 350
Alexa Fluor 405
Alexa Fluor 488
Alexa Fluor Plus 555
Rhodamine
Ex/Em (nm)
346/442
405/450
496/519
555/565
540/565
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing
Yes
Yes
Yes
Yes
Yes
Live cells
No
No
No
No
No
Fixed cells
Yes
Yes
Yes
Yes
Yes
Fixable
Yes
Yes
Yes
Yes
Yes
Platform
Imaging
Imaging
Imaging
Imaging
Imaging
Format
300 units
300 units
300 units
300 units
300 units
Cat. No. A30104
  Alexa Flour 568 Phalloidin Alexa Fluor Plus 750 Phalloidin
Readout
High-performance fluorescent label with stable signal, resistant to photobleaching
Range
F-actin 
Common filter set
Texas Red
Texas Red
Cy5
Cy5.5
Cy7
Labels
Alexa Fluor 568
Alexa Fluor 594
Alexa Fluor Plus 647
Alexa Fluor 680
Alexa Fluor 750
Ex/Em (nm)
578/600
590/617
650/665
679/702
758/784
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing Yes
Yes
Yes
Yes
Yes
Live cells No
No
No
No
No
Fixed cells Yes
Yes
Yes
Yes
Yes
Fixable Yes
Yes
Yes
Yes
Yes
Platform Imaging
Imaging
Imaging
Imaging
Imaging
Format 300 units
300 units
300 units
300 units
300 units
Cat. No. A12380
 
Readout
Live-cell stain
Live-cell stain
Target
β-tubulin
β-tubulin
Common filter set
FITC
Cy5/Deep Red
Labels
Oregon Green
Deep Red Fluorophore
Ex/Em (nm)
494/522
652/669
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing
Yes
Yes
Live cells
Yes
Yes
Fixed cells
No
No
Fixable
No
No
Platform
Imaging
Imaging
Format
1 kit
1 kit
Cat. No.
 
Readout
Live or fixed cell stain
Live or fixed cell stain
Live or fixed cell stain
Range
F-actin
F-actin
F-actin
Common filter set
FITC
TRITC/RFP
Cy5/Deep Red
Labels
Green Fluorophore
Orange Fluorophore
Deep Red Fluorophore
Ex/Em (nm)
503/512
545/570
652/669
Signal-to-noise ratio
Photostability
Bibliography
Multiplexing
Yes
Yes
Yes
Live cells
Yes
Yes
Yes
Fixed cells
Yes
Yes
Yes
Fixable
Yes
Yes
Yes
Platform
Imaging 
Imaging 
 Imaging
Format
1 vial or 5 vial
1 vial or 5 vial
1 vial or 5 vial
Cat. No.