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Fluorescent Protein Cellular Labels: Cellular Lights™ Reagents

Cellular Lights™ reagents consist of ready-to-use baculovirus suspensions carrying fluorescent protein expression construct targeted to specific subcellular structures.  These reagents provide a simple and effective method for introducing targeted intracellular labels within living cells.  Simply add the virus suspension to your cells, incubate overnight, and you’re ready to image your cells.

Advantages of labeling with Cellular Lights™ reagents

    • No lipids, chemical stains, or other potentially harmful treatments are required.
    • No need to optimize transduction conditions more than once per cell type.
    • Multiplex with cellular stains or other Organelle Lights™ or Cellular Lights™ reagents.
    • Efficient transduction of mammalian cell lines, including primary and stem cells.

    Cellular Lights™ reagents come in a variety of colors and targets (Table 1), including MAP4, actin and tubulin, for convenient multiplexing and co-localization studies.  

Overview of BacMam delivery technology



  • Figure 1.     		  Created as ready-to-use BacMam expression and delivery reagents (figure 1), the efficient transduction, robust expression and retained cellular function of these fusion proteins gives highly specific labeling for tools. Easily study dynamic changes, colocalization and signaling events in living cells. All Cellular Lights™ are compatible with fixed cell staining and high content analysis. For enhancement of the fluorescent protein signal in fixed samples, use one of our many anti-GFP antibodies.

    Visit www.invitrogen.com/BacMam for additional information on the BacMam delivery system, compatible cells and other BacMam reagents.

Table 1: Cellular Lights™ Reagents.

                                      
Cellular  Lights™ GFP YFP RFP
(488/510) (514/528) (555/584)
Actin C10126
C10127
CSF1r C10078

EXOC1
C10079
Histone 2B
 C10128
C10129
MAP4 C10105
C10140
TalinC10323
C10324
Tubulin C10106
C10112
Null virus (control)
C10130
Note:  Different cell types will have different transfection efficiencies with baculovirus -based technologies.  Certain cell types, such as macrophages and other hematopoiteic cells, cannot be transfected with baculovirus. See the varieties of cells that have been tested with Baculovirus Technology.

 

Cellular Lights™ and Organelle Lights™ Images

Cellular Lights™ Talin-GFP and Cellular Lights™ Actin RFP

HeLa cells were transduced with Cellular Lights™ Talin-GFP and co-transduced with Cellular Lights™ Actin RFP. Imaging was performed on live cells using a Delta–Vision® Core microscope and standard FITC/TRITC filter sets.
Cellular Lights™ Talin-RFP and Organelle Lights™ Mitochondria-GFP

HeLa cells were transduced with Cellular Lights™ Talin-RFP and Organelle Lights™ Mitochondria-GFP and Hoechst 33342 (1 ug/ml). Imaging was performed on live cells using a Delta–Vision® Core microscope and standard DAPI/FITC/TRITC filter sets.

Cellular Lights™ Videos

Time lapse movie of U-2 OS cells transduced with Cellular Lights™ Actin-RFP and Cellular Lights™ MAP4-GFP co-stained with Hoechst 33342

  U-2 OS cells were transduced with Cellular Lights™ Actin-RFP and Cellular Lights™ MAP4-GFP and imaged the next day. Cells were washed in HBSS (Gibco) and incubated with 1mg/ml Hoechst 33342 and washed with HBSS. Cells were kept in a climate controlled chamber and imaged with a 40X objective. Fluorescence was detected using a Delta–Vision® Core microscope and the appropriate filters (DAPI/FITC/TRITC). Cytochalsin D was applied to the bath at a final concentration of 10 mM. Images were taken every 20 seconds for 30 minutes. Note the disruption of actin filaments (red) whereas despite the change in cell shape microtubules are still clearly visible (green).
Time lapse movie of U-2 OS cells were transduced with Cellular Lights™ Histone 2B-RFP and Cellular Lights™ MAP4-GFP

  U-2 OS cells were transduced with Cellular Lights™ Histone 2B-RFP and Cellular Lights™ MAP4-GFP and imaged the next day. Cells were imaged in McCoys media. Cells were kept in a climate controlled chamber and imaged with a 40X objective, images were collected every 5 minutes for 7 hours and 35 minutes. Fluorescence was detected using a Delta–Vision® Core microscope and the appropriate filters (FITC/TRITC).
Time lapse movie of U-2 OS cells transduced with Cellular Lights™ Actin-RFP and Organelle Lights™ Plasma Membrane-GFP

  U-2 OS cells were transduced with Cellular Lights™ Actin-RFP and Organelle Lights™ Plasma Membrane-GFP and imaged the next day. Cells were imaged in McCoys media. Cells were imaged with a 40X objective. Images were acquired at 30 second intervals over a period of 45 minutes using a Delta–Vision® Core microscope and the appropriate filters (FITC/TRITC).
Time lapse movie of U-2 OS cells transduced with Cellular Lights™ Actin-RFP and Cellular Lights™ Tubilin-GFP

  U-2 OS cells were transduced with Cellular Lights™ Actin-RFP and Cellular Lights™ Tubilin-GFP and imaged the next day. Cells were imaged in McCoys media with a 40X objective and acquired at 5 minute intervals over a period of 9 hours. Fluorescence was detected using a Delta–Vision® Core microscope and the appropriate filters (FITC/TRITC).

BacMam Technology Overview

  • The BacMam system allows you to run the assays you need in relevant cellular models quickly with high quality results.  The BacMam technology offers the following key advantages.  

    • Efficient transduction of mammalian cell lines, including primary cells and stem cells
    • Safety (non-replicating in mammalian cells) and lack of observable cytopathic effect
    • Frozen storage of pre-transduced cells generates assay-ready cells
    • Easy modulation of gene expression by varying transduction amounts
    • Reproducible co-transduction and expression of multi-component complexes at the right stoichiometry

    Depending on transduction efficiency, cell type and cell division rate, the transgene remains detectable from 5 to 14 days.