Table 15.6 Fluorescent detection reagents for imaging mitochondria and lysosomes.
| Organic Dyes
(e.g., MitoTracker and LysoTracker dyes)
| BacMam-Based Fluorescent Proteins
(e.g., CellLight reagents)
|How they work||Positively charged MitoTracker dyes localize to actively respiring mitochondria; weakly basic LysoTracker dyes accumulate in compartments with low pH.||Combine targeting sequence–fluorescent protein fusion with the transduction efficiency of BacMam to label organelles independently of function (i.e., pH, mitochondrial membrane potential).||Recognize specific target of interest (e.g., LAMP1, a lysosomal protein).|
|Applications||Live-cell imaging applications; fixable and thus compatible with antibody-based imaging applications.*||Live-cell imaging applications; fixable and thus compatible with antibody-based imaging applications.*†||Imaging fixed cells or tissue. Compatible with labeling with MitoTracker, LysoTracker, and CellLight reagents.|
|Typical workflow||Incubate cells with the MitoTracker or LysoTracker reagent for approximately 5–30 min.||The ready-to-use CellLight reagent is added to live cells, followed by an overnight incubation to allow for protein expression.||Cells are fixed and permeabilized, then incubated with the antibody for labeling, and visualized with a fluorescently labeled secondary antibody.‡|
|* Please consult the product manual or contact technical service for additional information on the fixability of these reagents. † With fluorescent protein constructs, anti-GFP or anti-RFP antibodies can be used to amplify fluorescence signals. ‡ Secondary antibody is required if the primary antibody is not directly labeled.|
For Research Use Only. Not for use in diagnostic procedures.