title: Multi-color quantum dot tracking using a high-speed hyperspectral line-scanning microscope.
abstract: Many cellular signaling processes are initiated by dimerization or oligomerization of membrane proteins. However, since the spatial scale of these interactions is below the diffraction limit of the light microscope, the dynamics of these interactions have been difficult to study on living cells. We have developed a novel high-speed hyperspectral microscope (HSM) to perform single particle tracking of up to 8 spectrally distinct species of quantum dots (QDs) at 27 frames per second. The distinct emission spectra of the QDs allows localization with ∼10 nm precision even when the probes are clustered at spatial scales below the diffraction limit. The capabilities of the HSM are demonstrated here by application of multi-color single particle tracking to observe membrane protein behavior, including: 1) dynamic formation and dissociation of Epidermal Growth Factor Receptor dimers; 2) resolving antigen induced aggregation of the high affinity IgE receptor, FcεR1; 3) four color QD tracking while simultaneously visualizing GFP-actin; and 4) high-density tracking for fast diffusion mapping.
authors.affiliation: Department of Pathology and Cancer Research and Treatment Center, University of New Mexico, Albuquerque, New Mexico, United States of America.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Cutler
authors.forename: Michael D
authors.initials: MD
authors.lastname: Malik
authors.forename: Sheng
authors.initials: S
authors.lastname: Liu
authors.forename: Jason M
authors.initials: JM
authors.lastname: Byars
authors.forename: Diane S
authors.initials: DS
authors.lastname: Lidke
authors.forename: Keith A
authors.initials: KA
authors.lastname: Lidke
mesh_terms: D045744:Cell Line, Tumor; D003116:Color; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods; D045663:Quantum Dots; D015203:Reproducibility of Results; D015398:Signal Transduction
chemical_list: D008565:Membrane Proteins