Multipurpose analysis using second harmonic generating nanoprobes
First Claim
1. A second harmonic generating (SHG) nanoprobe system for nucleotide sequencing comprising:
- a plurality of distinct SHG nanoprobes, each of said distinct SHG nanoprobes having no inversion symmetry, the number of such distinct SHG nanoprobes being sufficient such that each distinct nucleotide type to be sequenced has a distinct SHG nanoprobe assigned thereto; and
an external excitation source for sequentially radiating each of a plurality of nucleotides to be sequenced over at least two distinct excitation wavelengths, wherein the SHG nanoprobes are selected such that the radiation generated by each of the SHG nanoprobes when said nanoprobes are excited at the at least two excitation wavelengths by the external excitation source creates an intensity ratio unique to each said distinct SHG nanoprobe when measured over at least two distinct emission wavelengths;
a detector configured to detect the sequential emissions of the plurality of distinct SHG nanoprobes at the at least Iwo distinct emission wavelengths; and
an analyzer for comparing the measured intensities from the sequential emissions at the at least two distinct emission wavelengths to determine a measured intensity ratio, and comparing said measured intensity ratio against a known standard for each of the distinct SHG nanoprobes, thereby sequentially determining each nucleotide in a target sequence.
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Abstract
Second harmonic nanoprobes for multipurpose imaging of samples and a method of using such probes to monitor nucleotide sequencing in a Multi-SHG Detection Imaging (MSDI) modality and to monitor external electric field using voltage sensitive second harmonic generating (SHG) nanoprobes are provided. The SHG nanoprobes are comprised of various kinds of nanocrystals that do not possess an inversion symmetry and therefore are capable of generating second harmonic signals that can then be detected by conventional two-photon microscopy for in vivo imaging of biological processes and structures such as cell signaling, neuroimaging, protein conformation probing, DNA conformation probing, gene transcription, virus infection and replication in cells, protein dynamics, tumor imaging and cancer therapy evaluation and diagnosis as well as quantification in optical imaging for a wide-range of biological and non-biological processes and devices.
19 Citations
17 Claims
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1. A second harmonic generating (SHG) nanoprobe system for nucleotide sequencing comprising:
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a plurality of distinct SHG nanoprobes, each of said distinct SHG nanoprobes having no inversion symmetry, the number of such distinct SHG nanoprobes being sufficient such that each distinct nucleotide type to be sequenced has a distinct SHG nanoprobe assigned thereto; and an external excitation source for sequentially radiating each of a plurality of nucleotides to be sequenced over at least two distinct excitation wavelengths, wherein the SHG nanoprobes are selected such that the radiation generated by each of the SHG nanoprobes when said nanoprobes are excited at the at least two excitation wavelengths by the external excitation source creates an intensity ratio unique to each said distinct SHG nanoprobe when measured over at least two distinct emission wavelengths; a detector configured to detect the sequential emissions of the plurality of distinct SHG nanoprobes at the at least Iwo distinct emission wavelengths; and an analyzer for comparing the measured intensities from the sequential emissions at the at least two distinct emission wavelengths to determine a measured intensity ratio, and comparing said measured intensity ratio against a known standard for each of the distinct SHG nanoprobes, thereby sequentially determining each nucleotide in a target sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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Specification