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Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
Published on: December 9, 2013
Two-photon fluorescence imaging super-enhanced by multishell nanophotonic particles, with application to subcellular
Aniruddha Ray1, Yong-Eun Koo Lee, Gwangseong Kim
1BioPhysics, University of Michigan, 930 N. University Ave. Ann Arbor, MI 48109, USA.
Small (Weinheim an Der Bergstrasse, Germany)
|April 21, 2012
Summary
Researchers developed a new nanophotonic method to boost two-photon fluorescence signals in live cells. This technique uses noble metal nanospheres to enhance signals up to 20 times, enabling clearer biological imaging.
Area of Science:
- Nanophotonics
- Biomedical Imaging
- Fluorescence Microscopy
Background:
- Two-photon fluorescence microscopy offers advantages like deeper tissue penetration and reduced photobleaching.
- Enhancing fluorescence signals is crucial for improving image quality and sensitivity in biological samples.
Purpose of the Study:
- To present a novel nanophotonic method for significantly enhancing two-photon fluorescence signals.
- To demonstrate this enhancement within a biological system, specifically live cells.
Main Methods:
- Utilizing noble metal nanospheres to generate second harmonic (SH) light, which further enhances fluorescence.
- Placing dye molecules in the near-field of nanospheres to leverage metal-enhanced fluorescence and SH effects.
- Employing multishell hydrogel nanoparticles with a silver core, citrate capping, pH indicator dye, and polyacrylamide cladding.
Main Results:
- Achieved up to a 20-fold enhancement in the two-photon fluorescence of an indicator dye.
- Successfully demonstrated two-photon fluorescence enhancement within live cells for the first time.
- Observed that enhanced signal includes one-photon processes from SH generation.
Conclusions:
- The novel nanophotonic method effectively enhances two-photon fluorescence signals in biological systems.
- This technique preserves the key benefits of two-photon microscopy, including deep penetration and low cellular damage.
- Offers a promising approach for advanced bioimaging applications requiring high sensitivity.
