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Total internal reflection fluorescence imaging using an upconverting cover slip for multicolour evanescent excitation

Morgan, CG and Mitchell, A 2006, 'Total internal reflection fluorescence imaging using an upconverting cover slip for multicolour evanescent excitation' , Journal of Microscopy, 222 (1) , pp. 48-57.

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Abstract

Total internal reflection fluorescence microscopy is well known as a means of studying surface-bound structures in cell biology. It is usually measured either by coupling a light source to the sample using a prism or with a special objective where light passing through the periphery of the lens illuminates the contact region beyond the critical angle. In this study we present a new and simple approach to total internal reflection fluorescence microscopy where the sample is mounted on a cover slip prepared from a high-index upconverting glass-ceramic. Excitation of the cover slip with a low-cost near-infrared laser diode generates intense narrow-band visible emission within the cover slip, some of which is totally internally reflected. This emission gives rise to an evanescent wave at the interface and hence can excite surface-bound fluorescent species. Depending on the excitation conditions the cover slip can generate violet, green and red emission and hence can excite a wide range of fluorescent labels. Fluorescence emission from the sample can be detected in spectral regions where the direct emission from the cover slip is very weak. The advantages and limitations of the technique are discussed in comparison with conventional total internal reflection fluorescence microscopy measurements and prospects for novel total internal reflection fluorescence microscopy geometries are considered.

Item Type: Article
Themes: Subjects / Themes > Q Science > Q Science (General)
Subjects outside of the University Themes
Schools: Colleges and Schools > College of Science & Technology
Colleges and Schools > College of Science & Technology > School of Environment and Life Sciences
Colleges and Schools > College of Science & Technology > School of Environment and Life Sciences > Biomedical Research Centre
Journal or Publication Title: Journal of Microscopy
Publisher: Blackwell Publishing
Refereed: Yes
ISSN: 00222720
Depositing User: H Kenna
Date Deposited: 08 Aug 2007 10:19
Last Modified: 20 Aug 2013 16:45
URI: http://usir.salford.ac.uk/id/eprint/162

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