Fluorescence lifetime imaging (FLI) in real-time - a new technique in photosynthesis research

Title:

Fluorescence lifetime imaging (FLI) in real-time - a new technique in photosynthesis research

Creator:

Holub, O., Seufferheld, M. J., Gohlke, C., Govindjee, and Clegg, R. M.

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:14f8c1b1-ebe4-4284-818b-994c1b18f33c
uuid:14f8c1b1-ebe4-4284-818b-994c1b18f33c
issn:0300-3604
doi:10.1023/A:1012465508465

Subject:

Arabidopsis, Chlamydomonas, FLIM, frequency domain, homodyne, microscopy, modulation, phase, photosystems 1 and 2, stress, time domain, and Zea

Type:

model:article and TEXT

Format:

bez média and svazek

Description:

We describe an instrument that allows the rapid measurement of fluorescence lifetime-resolved images of leaves as well as sub-cellular structures of intact plants or single cells of algae. Lifetime and intensity fluorescence images can be acquired and displayed in real time (up to 55 lifetime-resolved images per s). Our imaging technique therefore allows rapid measurements that are necessary to determine the fluorescence lifetimes at the maximum (P level) fluorescence following initial illumination during the chlorophyll (Chl) a fluorescence transient (induction) in photosynthetic organisms. We demonstrate the application of this new instrument and methodology to measurements of: (1) Arabidopsis thaliana leaves showing the effect of dehydration on the fluorescence lifetime images; (2) Zea mays leaves showing differences in the fluorescence lifetimes due to differences in the bundle sheath cells (having a higher amount of low yield photosystem 1) and the mesophyll cells (having a higher amount of high yield photosystem 2); and (3) single cells of wild type Chlamydomonas reinhardtii and its non-photochemical quenching mutant NPQ2 (where the conversion of zeaxanthin to violaxanthin is blocked), with NPQ2 showing lowered lifetime of Chl a fluorescence. In addition to the lifetime differences referred to in (1) and (2), structural dependent heterogeneities in the fluorescence lifetimes were generally observed when imaging mesophyll cells in leaves. and O. Holub ... [et al.].

Language:

Multiple languages

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Coverage:

581-599

Source:

Photosynthetica | 2000 Volume:38 | Number:4

Harvested from:

CDK

Metadata only:

false