The concept of using nanoparticles as a local sensor or probe enables important developments in medicine, molecular and cellular biology. Observing the entrance process of a particle into cells in real time, in 3D and without perturbation of the biological specimen is a dream of life science researchers.
Existing technologies such as epifluorescence, TIRF, LSM and video particle tracking have drawbacks. Labeling is time consuming and causes perturbation in single molecule experiments. The ideal resolution in confocal microscopy (CLSM) is ~300nm in the X and Y and ~500nm in the Z axis.
Fluorescence is not label free and has poor performance for processes occurring on cell membranes. TIRF measures only the first 200nm from the surface of the cell and gives no access to the whole cell volume. Particle tracking by video microscopy is only a 2D technique with best resolution of 15nm.
The answer is JPK Instruments’ NanoTracker, a live cell imaging technique available from Scitech with high temporal and spatial resolution applied to non labeled nanoparticles. It traps and tracks particles from several µm down to 30nm with the ability to control, manipulate and observe samples from vesicles to whole cells in real time with nanometer precision.
NanoTracker technology provides precisely quantifiable and reproducible measurements of particle/ cell interactions. The system delivers precise information about single molecule mechanics and may also be used to determine mechanical characteristics such as adhesion, elasticity or stiffness on single molecules.
Key features of NanoTracker include
- Real-time data - essential for living cell studies
- Optical tweezers meets 3D particle tracking
- 3D real space tracking information with nanometer precision
- Use of biochemically modified nanoparticles
- No perturbation of the biological system
- Reproducible and calibrated data
- Quantitative results from mechanical properties to diffusion
The following are a few applications of NanoTracker
- Analysis of protein (un)folding and function
- Interaction studies of DNA/RNA, motor proteins, carbohydrates with <pN sensitivity
- Molecular mechanics (pulling, stretching)
- Cell membrane research such as lateral organization (e.g. lipid rafts) or trans-membrane processes, trafficking
- Cell-particle interaction
- Diffusion measurements
- Nano-toxicity and drug delivery
- Endocytosis, Phagocytosis
- Cellular assembly
- Local gene or drug delivery
- Viral and bacterial infection
- Bacterial entrance mechanism studies
Modes of NanoTracker include
- Trapping, arranging and sorting mode
- Real-time trajectory data mode
- 3D real space tracking (thermal scanning mode)
- Volume scanning mode
- Force spectroscopy
- Force ramp and force clamp
- Force mapping
- Environment control with gas, liquid and temp control