Checkout View Cart Items

Fluorescent Protein Cellular Labels: Organelle Lights™ Reagents

Organelle Lights™ reagents consist of ready-to-go baculovirus suspensions carrying a 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 Organelle 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.

    Organelle Lights™ reagents come in a variety of colors and targets (Table 1), including mitochondria, ER, and Golgi 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 gives highly specific labeling tools. Easily study dynamic changes, colocalization and signaling events in living cells. All Organelle 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: Organelle Lights™ Reagents.


                         
Organelle Lights™ CFPGFPOFPRFP
(440/480)(488/510)(548/565)(555/584)
Cytoplasm
O36227

Endoplasmic Reticulum
O36212O36223O36230
Endosomes
O10104
O36231
Golgi
O36215O36224O10098
Lysosomes
O36228
O10100
Mitochondria
O36210O36222O36229
Nuclear Envelope
O36213

NucleusO36218O36209 O10099
Peroxisome
O36211 
Plasma MembraneO36216O36214O36226O10139
Synaptophysin
C10080

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.

 

Organelle Lights™ Images

Organelle Lights™ Lysosomes-GFP and Organelle Lights™ Mitochondria RFP

 HeLa cells were transduced with Organelle Lights™ Lysosomes-GFP and Organelle Lights™ Mitochondria RFP along with 1ug/ml Hoechst 33342. Imaging was performed on live cells using a Delta–Vision® Core microscope and standard DAPI/FITC/TRITC filter sets.
Organelle Lights™ ER-RFP and MitoTracker® Deep Red

 HeLa cells were transduced with Organelle Lights™ ER-GFP. HeLa cells were co-stained with 50 nM MitoTracker® Deep Red FM and 1mg/ml Hoechst 33342. Imaging was performed on live cells using a Delta–Vision® Core microscope and standard DAPI/TRITC/Cy5 filter sets.
Organelle Lights™ Endosomes-GFP and Organelle Lights™ Lysosomes RFP
 HeLa cells were transduced with Organelle Lights™ Endosomes-GFP and Organelle Lights™ Lysosomes RFP along with 1ug/ml Hoechst 33342. Imaging was performed on live cells using a Delta–Vision® Core microscope and standard DAPI/FITC/TRITC filter sets.

Organelle Lights™ Videos

Time lapse movie of HeLa cells were transduced with Organelle Lights™ Mito-GFP

  HeLa cells were transduced with Organelle Lights™ Mito-GFP and imaged the following day in MEM media with a 40X objective. Images were acquired at 5 second intervals over a period of 2 minutes and detected using the appropriate filters (FITC) on a Delta–Vision® Core microscope.
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 (12330). Cells were imaged with a 40X objective. Images were acquired at 30 second intervals over a period of 45 minutes using the appropriate filters (FITC/TRITC) on a Delta–Vision® Core microscope.

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.