Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy
Fluorophore excited state lifetime is a useful indicator of micro-environment in cellular optical molecular imaging. For quantitative sensing, precise lifetime determination is important, yet is often difficult to accomplish when using the experimental conditions favored by live cells. Here we repor...
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| Format: | Online |
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Optical Society of America
2010
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3410727/ |
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pubmed-3410727 |
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pubmed-34107272012-10-01 Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy Chang, Ching-Wei Mycek, Mary-Ann Research-Article Fluorophore excited state lifetime is a useful indicator of micro-environment in cellular optical molecular imaging. For quantitative sensing, precise lifetime determination is important, yet is often difficult to accomplish when using the experimental conditions favored by live cells. Here we report the first application of temporal optimization and spatial denoising methods to two-photon time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to improve lifetime precision in live-cell images. The results demonstrated a greater than five-fold improvement in lifetime precision. This approach minimizes the adverse effects of excitation light on live cells and should benefit FLIM applications to high content analysis and bioimage informatics. Optical Society of America 2010-04-09 /pmc/articles/PMC3410727/ /pubmed/20588712 http://dx.doi.org/10.1364/OE.18.008688 Text en ©2010 Optical Society of America http://creativecommons.org/licenses/by-nc-nd/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Unported License, which permits download and redistribution, provided that the original work is properly cited. This license restricts the article from being modified or used commercially. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Chang, Ching-Wei Mycek, Mary-Ann |
| spellingShingle |
Chang, Ching-Wei Mycek, Mary-Ann Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| author_facet |
Chang, Ching-Wei Mycek, Mary-Ann |
| author_sort |
Chang, Ching-Wei |
| title |
Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| title_short |
Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| title_full |
Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| title_fullStr |
Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| title_full_unstemmed |
Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| title_sort |
precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy |
| description |
Fluorophore excited state lifetime is a useful indicator of micro-environment in cellular optical molecular imaging. For quantitative sensing, precise lifetime determination is important, yet is often difficult to accomplish when using the experimental conditions favored by live cells. Here we report the first application of temporal optimization and spatial denoising methods to two-photon time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to improve lifetime precision in live-cell images. The results demonstrated a greater than five-fold improvement in lifetime precision. This approach minimizes the adverse effects of excitation light on live cells and should benefit FLIM applications to high content analysis and bioimage informatics. |
| publisher |
Optical Society of America |
| publishDate |
2010 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3410727/ |
| _version_ |
1611547276927303680 |