Cell Tracking

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.

Select products for cell tracking

Short-Term Cell Tracking

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. 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.

Select short-term cell tracking products

Rat basophilic leukemia (RBL) cells labeled in suspension with CellTracker™ Blue CMAC, CellTracker™ Green CMFDA or CellTracker™ Orange CMTMR prior to plating.

Long-Term Cell Tracking

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. 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.

Select long-term cell tracking products

Multiplexing Qdot® probes, dyes, and fluorescent proteins. U2-OS cells were transduced with CellLight® Plasma Membrane-GFP (green), followed by labeling with Qtracker® 655 Cell Labeling Kit (magenta), and HCS NuclearMask™ Blue (blue).

Cell Tracking with Fluorescent Proteins

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. CellLight® fluorescent proteins can be expressed in a variety of cell types including numerous transformed cell lines, primary cells, stem cells and neurons.

Selection guide for cell tracking with expressed protein

Porcine primary skeletal muscle cell transduced with CellLight® Nucleus-GFP fluorescent proteins.

Cell Tracking Selection Guides

Short-term tracking requires rapid loading and typical retention for 3–6 generations or 72 hours.

Properties

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
*In vivo*
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	Cy®5	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	GFP or FITC	565	1 kit	Q25031MP
Qtracker® 585 Cell Labeling Kit	TRITC-A	585	1 kit	Q25011MP
Qtracker® 605 Cell Labeling Kit	TRITC-A	605	1 kit	Q25001MP
Qtracker® 625 Cell Labeling Kit	Texas Red®	625	1 kit	A10198
Qtracker® 655 Cell Labeling Kit	Texas Red®	655	1 kit	Q25021MP
Qtracker® 705 Cell Labeling Kit	Cy®5	705	1 kit	Q25061MP
Qtracker® 800 Cell Labeling Kit	Cy®7	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.

	CellLight® Nucleus-RFP	CellLight® Nucleus-GFP	BacMam GFP Transduction Control
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
HCS
*In vivo*	Yes
Bibliography	Citations
Quantity	1 mL	1 mL	1 mL
Cat. No.	C10603	C10602	B10383

Short-Term Cell Tracking

Short-term tracking requires rapid loading and typical retention for 3–6 generations or 72 hours.

Properties

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
*In vivo*
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	Cy®5	630/650	20 x 15 µg	C34565

Long-Term Cell Tracking

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	GFP or FITC	565	1 kit	Q25031MP
Qtracker® 585 Cell Labeling Kit	TRITC-A	585	1 kit	Q25011MP
Qtracker® 605 Cell Labeling Kit	TRITC-A	605	1 kit	Q25001MP
Qtracker® 625 Cell Labeling Kit	Texas Red®	625	1 kit	A10198
Qtracker® 655 Cell Labeling Kit	Texas Red®	655	1 kit	Q25021MP
Qtracker® 705 Cell Labeling Kit	Cy®5	705	1 kit	Q25061MP
Qtracker® 800 Cell Labeling Kit	Cy®7	800	1 kit	Q25071MP

Cell Tracking With Fluorescent Proteins

Transduction with fluorescent proteins provides long-term tracking, but since not all cells are transformed some automated techniques are not supported.

	CellLight® Nucleus-RFP	CellLight® Nucleus-GFP	BacMam GFP Transduction Control
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
HCS
*In vivo*	Yes
Bibliography	Citations
Quantity	1 mL	1 mL	1 mL
Cat. No.	C10603	C10602	B10383

Resources
Molecular Probes® Handbook Fluorescent tracers of cell morphology—Section 14.1 Polar tracers—Section 14.3 Lipophilic tracers—Section 14.4 Dextran conjugates—Section 14.5 Protein conjugates—Section 14.7 Tracers for blood flow—Section 6.6	BioProbes® articles Tracking Cells Through Time and Space—Choose From Fluorescent Dyes, Qdot® Nanocrystals, and Targeted Fluorescent Proteins Going Viral—Fluorescent Probes to Image Viruses and Their Host Cell Interactions

On this page:

Short-Term Cell Tracking

Long-Term Cell Tracking

Cell Tracking with Fluorescent Proteins

Cell Tracking Selection Guides

Properties

Selection Guide

Properties

Selection Guide

Properties

Selection Guide

Properties

Selection Guide

Resources