Cell movement and location studies require specialized probes that are nontoxic to living cells and are available in a range of fluorescent colors to match instrument lasers and filters and to accommodate co-staining with antibodies or other cell analysis probes. Choose passively loaded dyes for short- and long-term tracking or fluorescent fusion proteins for cell tracking over 5 or more days.
- Rapid loading (typically 15 min to 1 hr)
- Short-term tracking (typically 3–6 generations)
- Moderate resistance to photobleaching
- Moderate brightness
- Signal distribution throughout the cell
The simplest processes for monitoring cell movement and location use tracking probes that pass through the membrane into the cell and become membrane-impermeant after loading. Ideal tracking dyes are passed to daughter cells for multiple generations but do not transfer to neighboring cells on contact. Invitrogen CellTracker dyes exhibit ideal properties—they are stable, nontoxic at working concentrations, well retained in cells, and brightly fluorescent at physiological pH. CellTracker dyes have a wide range of emission spectra for easy combination with antibody staining.
- Typically 1-hour cell loading
- Long-term tracking (typically 6–10 generations)
- High resistance to photobleaching
- Intense brightness
- Punctate distribution throughout the cell
Longer-term tracking of cell movement and location requires probes that are more resistant to photobleaching and are retained through more cycles of cell division. Invitrogen Qtracker probes provide intense, stable signals for long-term (6–10 generations) tracking. The eight distinct probe colors are resistant to photobleaching and provide intense punctate fluorescence within the cell, with different emissions from a single excitation for easy combination with antibody staining. Qtracker probes have a wide range of emission spectra for easy combination with antibody staining.
- Simple transduction protocol
- Overnight incubation
- Long-term tracking for up to 2 weeks or more than 10 generations
- Red or green fluorescent protein
- Can be multiplexed with fluorescent dyes or proteins
Transiently transduced cells express fluorescent fusion proteins at different locations within the cell for at least 5 days or up to 2 weeks in slow-growing populations. Labeled cells are easy to track, but not all cells within a population will express the fluorescent protein. Invitrogen CellLight fluorescent proteins can be expressed in a variety of cell types including numerous transformed cell lines, primary cells, stem cells and neurons.
Cell morphology and image segmentation
- Label nucleus and cytoplasm
- No wash steps required
- Multiple wavelength options
- Validated for automated imaging
Identifying cell boundaries is a key step in automated image acquisition and analysis for high-content screening (HCS) assays. Image segmentation involves separating objects of interest (such as cells) from background or other features not relevant to the analysis. It allows quantitation of morphological changes and items of interest in their cellular context, such as the translocation of biomarkers between different organelles. We offer Invitrogen HCS CellMasks for whole cell demarcation in a variety of wavelengths for multiplexing together with other structural and functional probes.
Cell tracking selection guides
Short-term tracking requires rapid loading and typical retention for 3–6 generations or 72 hours.
Properties
| CellTracker dyes | |
|---|---|
| Method of use | 15 minutes to 1 hour of cell loading |
| Readout | Internalized probe gives bright fluorescence throughout the cell |
| Range | Tracking extends for 3–6 generations or 72 hours |
| Compatibility | Multiplex easily with antibody labeling after fixing |
| Signal-to-noise |  |
| Photostability | |
| Fluorescence microscopy | Yes |
| HCS | Yes |
| Bibliography | Citations |
Selection Guide
| CellTracker dye | Common filter set | Ex/Em (nm) | Quantity | Cat. No. | |
|---|---|---|---|---|---|
| CellTracker Blue CMAC (7-amino-4-Chloromethylcoumarin) | CMAC | DAPI | 353/466 | 5 mg | C2110 |
| CellTracker Blue CMF2HC (4-chloromethyl-6,8-difluoro-7-hydroxycoumarin) | Blue CMF2HC | DAPI | 371/464 | 5 mg | C12881 |
| CellTracker Violet BMQC (2,3,6,7-tetrahydro-9- bromomethyl-1H,5H-quinolizino( 9,1-gh)coumarin) | Violet BMQC | DAPI | 415/516 | 5 x 0.1 mg | C10094 |
| CellTracker Green CMFDA (5-chloromethylfluorescein diacetate) | Green CMFDA | FITC | 492/517 | 20 x 50 µg | C7025 |
| CellTracker Orange CMRA | Orange CMRA | TRITC | 548/576 | 20 x 50 µg | C34551 |
| CellTracker CM-DiI | CM-DiI | TRITC | 553/570 | 20 x 50 µg | C7000 |
| CellTracker Red CMTPX | CMTPX | Rhodamine | 577/602 | 20 x 50 µg | C34552 |
| CellTracker Deep Red | Deep Red | Cy5 | 630/650 | 20 x 15 µg | C34565 |
Long-term tracking requires longer photostability and typical retention for 6–10 generations; loading time is less critical to experimental protocols.
Properties
| Qtracker 525 Cell Labeling Kit | |
|---|---|
| Method of use | 1-hour cell loading using a targeting peptide |
| Readout | Internalized probe gives intense punctate fluorescence within the cell |
| Range | Tracking extends for 6–10 generations or 5–14 days |
| Fixablility | Image cells live or after fixing |
| Compatibility | Multiplex easily with antibody labeling |
| Signal-to-noise |  |
| Photostability | |
| Laser excitation | Excited optimally with UV up to 488 nm (also every wavelength below the emission wavelength) |
| Fluorescence microscopy | Yes |
| HCS | Yes |
| In vivo | Yes |
| Bibliography | Citations |
Selection Guide
Common filter set
|
Emission max (nm)
|
Quantity
| Cat. No. | |
|---|---|---|---|---|
| Qtracker 525 Cell Labeling Kit | GFP or FITC | 525 | 1 kit | Q25041MP |
| Qtracker 565 Cell Labeling Kit | 565 | 1 kit | Q25031MP | |
| Qtracker 585 Cell Labeling Kit | TRITC-A | 585 | 1 kit | Q25011MP |
| Qtracker 605 Cell Labeling Kit | 605 | 1 kit | Q25001MP | |
| Qtracker 625 Cell Labeling Kit | Texas Red | 625 | 1 kit | A10198 |
| Qtracker 655 Cell Labeling Kit | 655 | 1 kit | Q25021MP | |
| Qtracker 705 Cell Labeling Kit | Cy5 | 705 | 1 kit | Q25061MP |
| Qtracker 800 Cell Labeling Kit | Cy7 | 800 | 1 kit | Q25071MP |
Transduction with fluorescent proteins provides long-term tracking, but since not all cells are transformed some automated techniques are not supported.
| Method of use | Transduction of cell line with fluorescent protein requires overnight incubation | ||
| Readout | Up to 90% of cells express fluorescent protein in respective cell compartments | ||
| Range | Expressed for ≥10 cell generations; 5 days to 2 weeks of tracking | ||
| Dye |
RFP
|
GFP
|
GFP
|
| Ex/Em (nm) |
555/584
|
488/510
|
488/510
|
| Fixability | | ||
| Multiplexing | | ||
| Signal-to-noise | | ||
| Photostability | | ||
| Location |
Nucleus
|
Cytoplasm
| |
| Fluorescence microscopy |
Yes
| ||
| In vivo |
Yes
| ||
| Bibliography |
Citations
| ||
| Quantity |
1 mL
|
1 mL
|
1 mL
|
| Cat. No. |
C10603
|
C10602
|
B10383
|
| HCS CellMask Red | HCS CellMask Deep Red | ||||
|---|---|---|---|---|---|
| Readout |
Stains nucleus and cytoplasm for whole-cell demarcation
| ||||
| Common filter set | DAPI |
FITC
|
RFP
|
Texas Red
| Cy5 |
| Ex/Em (nm) |
346⁄442
|
493/516
|
556/572
|
588⁄612
|
650⁄665
|
| Signal-to-noise ratio |  |  | | | |
| Photostability | | | | | |
| Multiplexing |
Yes
|
Yes
|
Yes
|
Yes
|
Yes
|
| Live cells |
No
|
No
|
No
|
No
|
No
|
| Fixed cells |
Yes
|
Yes
|
Yes
|
Yes
|
Yes
|
| Platform |
HCS
|
HCS
|
HCS
|
HCS
|
HCS
|
| Protocol | High-content analysis | ||||
| Format |
250 µg/10 plates
|
250 µg/10 plates
|
250 µg/10 plates
|
250 µg/10 plates
|
250 µg/10 plates
|
| Cat. No. | |||||
Short-term tracking requires rapid loading and typical retention for 3–6 generations or 72 hours.
Properties
| CellTracker dyes | |
|---|---|
| Method of use | 15 minutes to 1 hour of cell loading |
| Readout | Internalized probe gives bright fluorescence throughout the cell |
| Range | Tracking extends for 3–6 generations or 72 hours |
| Compatibility | Multiplex easily with antibody labeling after fixing |
| Signal-to-noise | |
| Photostability | |
| Fluorescence microscopy | Yes |
| HCS | Yes |
| Bibliography | Citations |
Selection Guide
| CellTracker dye | Common filter set | Ex/Em (nm) | Quantity | Cat. No. | |
|---|---|---|---|---|---|
| CellTracker Blue CMAC (7-amino-4-Chloromethylcoumarin) | CMAC | DAPI | 353/466 | 5 mg | C2110 |
| CellTracker Blue CMF2HC (4-chloromethyl-6,8-difluoro-7-hydroxycoumarin) | Blue CMF2HC | DAPI | 371/464 | 5 mg | C12881 |
| CellTracker Violet BMQC (2,3,6,7-tetrahydro-9- bromomethyl-1H,5H-quinolizino( 9,1-gh)coumarin) | Violet BMQC | DAPI | 415/516 | 5 x 0.1 mg | C10094 |
| CellTracker Green CMFDA (5-chloromethylfluorescein diacetate) | Green CMFDA | FITC | 492/517 | 20 x 50 µg | C7025 |
| CellTracker Orange CMRA | Orange CMRA | TRITC | 548/576 | 20 x 50 µg | C34551 |
| CellTracker CM-DiI | CM-DiI | TRITC | 553/570 | 20 x 50 µg | C7000 |
| CellTracker Red CMTPX | CMTPX | Rhodamine | 577/602 | 20 x 50 µg | C34552 |
| CellTracker Deep Red | Deep Red | Cy5 | 630/650 | 20 x 15 µg | C34565 |
Long-term tracking requires longer photostability and typical retention for 6–10 generations; loading time is less critical to experimental protocols.
Properties
| Qtracker 525 Cell Labeling Kit | |
|---|---|
| Method of use | 1-hour cell loading using a targeting peptide |
| Readout | Internalized probe gives intense punctate fluorescence within the cell |
| Range | Tracking extends for 6–10 generations or 5–14 days |
| Fixablility | Image cells live or after fixing |
| Compatibility | Multiplex easily with antibody labeling |
| Signal-to-noise | |
| Photostability | |
| Laser excitation | Excited optimally with UV up to 488 nm (also every wavelength below the emission wavelength) |
| Fluorescence microscopy | Yes |
| HCS | Yes |
| In vivo | Yes |
| Bibliography | Citations |
Selection Guide
Common filter set
|
Emission max (nm)
|
Quantity
| Cat. No. | |
|---|---|---|---|---|
| Qtracker 525 Cell Labeling Kit | GFP or FITC | 525 | 1 kit | Q25041MP |
| Qtracker 565 Cell Labeling Kit | 565 | 1 kit | Q25031MP | |
| Qtracker 585 Cell Labeling Kit | TRITC-A | 585 | 1 kit | Q25011MP |
| Qtracker 605 Cell Labeling Kit | 605 | 1 kit | Q25001MP | |
| Qtracker 625 Cell Labeling Kit | Texas Red | 625 | 1 kit | A10198 |
| Qtracker 655 Cell Labeling Kit | 655 | 1 kit | Q25021MP | |
| Qtracker 705 Cell Labeling Kit | Cy5 | 705 | 1 kit | Q25061MP |
| Qtracker 800 Cell Labeling Kit | Cy7 | 800 | 1 kit | Q25071MP |
Transduction with fluorescent proteins provides long-term tracking, but since not all cells are transformed some automated techniques are not supported.
| Method of use | Transduction of cell line with fluorescent protein requires overnight incubation | ||
| Readout | Up to 90% of cells express fluorescent protein in respective cell compartments | ||
| Range | Expressed for ≥10 cell generations; 5 days to 2 weeks of tracking | ||
| Dye |
RFP
|
GFP
|
GFP
|
| Ex/Em (nm) |
555/584
|
488/510
|
488/510
|
| Fixability | | ||
| Multiplexing | | ||
| Signal-to-noise | | ||
| Photostability | | ||
| Location |
Nucleus
|
Cytoplasm
| |
| Fluorescence microscopy |
Yes
| ||
| In vivo |
Yes
| ||
| Bibliography |
Citations
| ||
| Quantity |
1 mL
|
1 mL
|
1 mL
|
| Cat. No. |
C10603
|
C10602
|
B10383
|
| HCS CellMask Red | HCS CellMask Deep Red | ||||
|---|---|---|---|---|---|
| Readout |
Stains nucleus and cytoplasm for whole-cell demarcation
| ||||
| Common filter set | DAPI |
FITC
|
RFP
|
Texas Red
| Cy5 |
| Ex/Em (nm) |
346⁄442
|
493/516
|
556/572
|
588⁄612
|
650⁄665
|
| Signal-to-noise ratio | | | | | |
| Photostability | | | | | |
| Multiplexing |
Yes
|
Yes
|
Yes
|
Yes
|
Yes
|
| Live cells |
No
|
No
|
No
|
No
|
No
|
| Fixed cells |
Yes
|
Yes
|
Yes
|
Yes
|
Yes
|
| Platform |
HCS
|
HCS
|
HCS
|
HCS
|
HCS
|
| Protocol | High-content analysis | ||||
| Format |
250 µg/10 plates
|
250 µg/10 plates
|
250 µg/10 plates
|
250 µg/10 plates
|
250 µg/10 plates
|
| Cat. No. | |||||
