Laser Scanning Up-Conversion Luminescence Microscopy for Imaging Cells Labeled with Rare-Earth Nanophosphors
- 6 January 2009
- journal article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 81 (3), 930-935
- https://doi.org/10.1021/ac802072d
Abstract
Because of the ability to selectively reveal the objects of interest with subcellular resolution, fluorescence microscopy provides widespread applications from basic biological research to clinical diagnosis. However, challenges still remain in reducing the degree of photobleaching and increasing the contrast between signal and noise. Herein, we found that rare-earth nanophosphors exhibit a unique up-conversion luminescence mechanism and imaging modality and developed a new three-dimensional visualization method of laser scanning up-conversion luminescence microscopy (LSUCLM) with little photobleaching and no background fluorescence, by introducing a reverse excitation dichroic mirror and the confocal pinhole technique. Moreover, we demonstrated the up-conversion emission imaging of thin films containing embedded rare-earth nanophosphors and cells multilabeled with the nanophosphors and organic dyes. These data show that LSUCLM not only shares noninvasive benefits and deep penetration of two-photon microscopy but also offers some distinct advantages, such as little photobleaching of both organic dyes and rare-earth nanophosphors, no background fluorescence from either endogenous fluorophores or colabeled fluorescent probes, and excellent compatibility with conventional confocal microscopy.Keywords
This publication has 32 references indexed in Scilit:
- Combined in Situ Atomic Force Microscopy- Infrared-Attenuated Total Reflection SpectroscopyAnalytical Chemistry, 2007
- A Highly Selective Fluorescence Turn-on Sensor for Cysteine/Homocysteine and Its Application in BioimagingJournal of the American Chemical Society, 2007
- Living Cells as Test TubesScience, 2006
- UV Fluorescence Lifetime Imaging Microscopy: A Label-Free Method for Detection and Quantification of Protein InteractionsAnalytical Chemistry, 2005
- Optical sectioning microscopyNature Methods, 2005
- Dynamic Multiphoton Imaging: A Live View from Cells to SystemsPhysiology, 2005
- Scanning Probe MicroscopyAnalytical Chemistry, 2004
- Nonlinear magic: multiphoton microscopy in the biosciencesNature Biotechnology, 2003
- Light Microscopy Techniques for Live Cell ImagingScience, 2003
- Two-photon tissue imaging: seeing the immune system in a fresh lightNature Reviews Immunology, 2002