# US 2014/0194314 A1 · Multi-Color Nanoscale Imaging Based On Nanoparticle Cathodoluminescence

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- Number: US 2014/0194314 A1
- Document ID: US20140194314A1
- Status: Published application
- Kind: A1
- Application number: 14/125,934
- Filed: 2012-06-13
- Published: 2014-07-10
- Earliest claimed: 2011-06-13
- Assignees: President and Fellows of Harvard College
- Inventors: Ronald Walsworth; Jeff Lichtman; Narayanan Kasthuri; David Glenn; Huiliang Zhang; Richard Schalek
- Web page: https://patentgrep.com/patent/US20140194314A1

## CPC

- G01N 23/2254: Measuring cathodoluminescence
- G01N 33/54373: {involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings}
- H01J 37/22: Optical {, image processing} or photographic arrangements associated with the tube
- H01J 37/244: Detectors; Associated components or circuits therefor
- G01N 2223/403: mapping with false colours
- G01N 2223/418: electron microscope
- G01N 2223/612: biological material
- H01J 2237/225: Displaying image using synthesised colours
- H01J 2237/2808: Cathodoluminescence
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- H01J: ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS (spark-gaps H01T; electric arc lamps H05B31/00; particle accelerators H05H)

## Abstract

Multi-color CL images of nanoparticle samples may be generated, by irradiating with a scanning electron beam a nanoparticle sample that containing a plurality of spectrally distinct optical emitters configured to generate CL light at respective different color channels, then detecting the CL light from the nanoparticles to generate multi-color NP-CL images of the nanoparticle sample. In some embodiments, SE (secondary electron) images of the sample may be acquire, substantially simultaneously with the acquisition of the CL images, so as to generate correlative NP-CL and SE images of the nanoparticle sample. In some embodiments, the nanoparticles may be surface-functionalized so that the nanoparticles selectively bind only to particular structures of interest.

## Claims

### Claim 1

A method comprising:
irradiating with a scanning electron beam a nanoparticle sample containing a plurality of nanoparticles, so as to cause emission of CL (cathodoluminescent) photons from the nanoparticle sample; wherein the nanoparticles comprise a plurality of spectrally distinct optical emitters configured to generate CL light at respective different color channels; and
detecting the CL light from the nanoparticles to generate multi-color NP (nanoparticle)-CL images of the nanoparticle sample.

### Claim 2 (depends on 1)

The method of claim 1, further comprising acquiring SE (secondary electron) images of the sample, substantially simultaneously with the acquisition of the CL images, so as to generate correlative NP-CL and SE images of the nanoparticle sample.

### Claim 3 (depends on 1)

The method of claim 1, wherein the nanoparticles comprise surface-functionalized nanoparticles, and further comprising the acts of:
surface functionalizing a plurality of nanoparticles so as to allow the nanoparticles to selectively bind only to a particular structure of interest;
depositing the surface-functionalized nanoparticles onto a substrate to create the nanoparticle sample.

### Claim 4 (depends on 3)

The method of claim 3, wherein the act of surface functionalizing the nanoparticles comprises coating the surface of each nanoparticle with a predetermined substance so that the nanoparticle becomes conjugated with a particular antibody.

### Claim 5 (depends on 2)

The method of claim 2, wherein the acts of acquiring the SE images of the sample and acquiring the multi-color CL images of the sample are performed substantially simultaneously.

### Claim 6 (depends on 2)

The method of claim 2, wherein the acts of acquiring the SE images of the sample and acquiring the multi-color CL images of the sample are performed using one integrated instrument.

### Claim 7 (depends on 1)

The method of claim 1, wherein the nanoparticles comprise nanodiamonds containing NV (nitrogen-vacancy) color centers.

### Claim 8 (depends on 1)

The method of claim 1, wherein the nanoparticles comprise LuAG:Ce (Cerium-doped Lutetium-Aluminum) nanophosphors.

### Claim 9 (depends on 1)

The method of claim 1, wherein the nanoparticles comprise nanodiamonds including band-A defects.

### Claim 10 (depends on 3)

The method of claim 3, wherein the particular structures of interest comprise synaptic vesicles.

### Claim 11

A system comprising:
an electron microscope configured to generate an image of a nanoparticle sample containing a plurality of nanoparticles, by irradiating the nanoparticle sample with electron beams;
wherein the nanoparticles comprise a plurality of spectrally distinct optical emitters configured to generate CL (cathodoluminescent) light at respective different color channels in response to the electron beams from the electron microscope; and
a CL detecting imaging system configured to detect the CL light from the nanoparticles and to generate multi-color CL images of the nanoparticle sample.

### Claim 12 (depends on 11)

The system of claim 11, wherein the electron microscope and the CL imaging system are integrated into a single instrument.

### Claim 13 (depends on 11)

The system of claim 11, wherein at least some of the nanoparticles are surface functionalized, so that each one of the surface-functionalized nanoparticles selectively binds only to a particular structure of interest.

### Claim 14 (depends on 13)

The system of claim 13, wherein the surface-functionalized nanoparticles comprise antibody-conjugated nanoparticles.

### Claim 15 (depends on 11)

The system of claim 11, wherein the electron microscope comprises one of: a SEM (scanning electron microscope); an STM (scanning tunneling microscope); an STEM (scanning transmission electron microscope) and a TEM (transmission electron microscope).

### Claim 16 (depends on 11)

The system of claim 11, wherein the nanoparticles comprise nanodiamonds containing NV (nitrogen-vacancy) color centers, and are configured to produce red cathodoluminescent light at a wavelength of about 620 nm.

### Claim 17 (depends on 11)

The system of claim 11, wherein the nanoparticles comprise LuAG:Ce (Cerium-doped Lutetium-Aluminum) nanophosphors, and are configured to produce green cathodoluminescent light at a wavelength of about 510 nm.

### Claim 18 (depends on 11)

The system of claim 11, wherein the nanoparticles comprise nanodiamonds including band-A defects, and are configured to produce blue cathodoluminescent light at at wavelength of about 420 nm.

### Claim 19 (depends on 13)

The system of claim 13, wherein the particular structures of interest comprise synaptic vesicles.