Ultrahigh Resolution Multicolor Colocalization of Single Fluoresecent Probes

Allows for ultra-high resolution fluorescence imaging that eliminates many of the limitations of current techniques.Nanometer accuracy.Allows for 3D scanning and multicolor imaging.Potential in vivo nanometer-resolution mapping and tracking of cellular components.

Genome mappingAnalysis of DNA binding proteinsFluorescence in situ hybridizationImmunocytologyImage cytometryIn vivo imagingAnalysis of protein-protein interactionsHigh throughput screeningData storage

UCLA investigators have developed a novel imaging and ultrahigh-resolution colocalization technique that can pinpoint the location of multiple distinguishable probes with nanometer accuracy and perfect registry. Based on sample-scanning confocal microscopy, it uses a single excitation laser and a closed-loop piezo-scanner that allows for nanometer accuracy steps. Also, this invention utilizes point-like fluorescent probes that can all be excited by the same laser wavelength but differ in their emission properties (i.e., semiconductor nanocrystals). Because all the probes are excited by the same laser aligned on the optical axis, chromatic aberrations are altogether eliminated. The fixed-excitation scheme also ensures the equivalence of each channel in the detection path. Finally, because each image is constructed pixel-by-pixel from the recorded signal of each channel, all the pixels corresponding to a given scanner position are in perfect registry. This approach will allow 3D scanning and multicolor imaging, opening the way to in vivo nanometer-resolution mapping and tracking of multiple cellular components.

Patent Number: US6844150B2
Application Number: US2001925100A
Inventor: Weiss, Shimon | Michalet, Xavier | Lacoste, Thilo D.
Priority Date: 24 Aug 2000
Priority Number: US6844150B2
Application Date: 8 Aug 2001
Publication Date: 18 Jan 2005
IPC Current: G02B002100
US Class: 435004 | 422050 | 4220681 | 42208205 | 42208208 | 43500612 | 435006
Assignee Applicant: The Regents of the University of California
Title: Ultrahigh resolution multicolor colocalization of single fluorescent probes
Usefulness: Ultrahigh resolution multicolor colocalization of single fluorescent probes
Summary: In biological and nanotechnological applications for colocalization of point-like molecular probes such as transfluospheres, malaria parasite proteins in membrane of infected red blood cells, RNAs, in vivo nanometer-resolution mapping and tracking of multiple cellular components, colocalization of semiconductor nanocrystals for ultra high resolution mapping of genes and DNA binding proteins, nucleic acids, organelles within living cells.
Novelty: Fluorescent species colocalization method in biological sample analysis, involves computing distance between geometric centers of point spread function excitation for two species

Biomedical

Medical Imaging

6844150

Background

There is a growing focus now on understanding how the fundamental cellular building blocks are organized and interact with each other. A tool is needed that can provide dynamic in vivo 3-dimensional microscopic pictures with nanometer resolution of individual molecules interacting with each other. 

Currently, fluorescence microscopy can provide detailed observations down to the single molecule level for in vitro experiments, and that single fluorophores can be detected in the membrane of living cells with good signal-to-noise ratios. However, it is uncertain whether this method can provide the required spatial and temporal resolution necessary for imaging molecular interactions in vivo. Although several advances have been made in improving the spatial resolution of optical microscopy, they all have their limitations. Some of these limitations include a limited ability to compensate for aberrations, a limited implementation for hydrated samples, constraints on sample size, and difficulty expanding to multi-color probes. Further, super-resolution approaches suffer from basic limitations of far-field optics such as spherical and chromatic aberrations. Although attempts have been made to correct these difficulties, none of these approaches perfectly corrects these imperfections. 

Related Materials

Ultrahigh-resolution multicolor colocalization of single fluorescent probes

Additional Technologies by these Inventors

• Protein-Protein Interactions as a Tool for Site-Specific Labeling of Proteins
• Method and Apparatus for Photon Arrival Time Interval Distribution (Paid) Analysis in Fluorescence Correlation Spectroscopy
• Bioactivation and Surface Properties Modulation of Inorganic Nanoparticles
• Membrane Insertion of Potential Sensing Nanorods

Tech ID/UC Case

21725/2003-500-0

Related Cases

2003-500-0

USA

Desktop View