Abstract

We demonstrate that picosecond time-gated fluorescence microscopy can be used to monitor subtle changes in the kinetics and spatial distribution of perturbations to the molecular and cellular structure of plant tissue caused by ultraviolet radiation. Single-molecule experiments on Photosystem II and chloroplast preparations give picosecond fluorescence decay kinetics that are similar to those obtained previously on bulk samples. For green plant leaves, localized and well-defined cellular structure is seen for normal material whereas relatively diffuse and non-specific features are seen after UV-irradiation indicating significant UV-induced rupture of the cellular structure. The changes in the chlorophyll fluorescence decay kinetics indicate uncoupling of chlorophyll molecules in the light-harvesting system leading to inhibition of energy reorganization and transfer in the antennae and subsequent exciton transfer to the reaction centers.

© 2002 Optical Society of America

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  1. G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
    [Crossref] [PubMed]
  2. J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
    [Crossref]
  3. K. Dowling, M. J. Dayel, M. J. Lever, P. M. W. French, J. D. Hares, and A. K. L. Dymoke-Bradshaw, “Fluorescence lifetime imaging with picosecond resolution for biomedical applications,” Opt. Lett. 23, 810–812 (1998).
    [Crossref]
  4. M. Straub and S. W. Hell, “Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope,” Appl. Phys. Lett. 73, 1769–1771 (1998).
    [Crossref]
  5. T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
    [Crossref]
  6. T. A. Roelofs, C. -H. Lee, and A. R. Holzwarth, “Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in Photosystem II alpha- and beta-units,” Biophys. J. 61, 1147–1163 (1992).
    [Crossref] [PubMed]
  7. V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
    [Crossref] [PubMed]
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    [Crossref]
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  10. M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).
  11. J. M. Anderson, Y. -I. Park, and W. S. Chow, “Photoinactivation and photoprotection of Photosystem II in nature”, Physiol. Plantarum 100, 214–223 (1997).
    [Crossref]
  12. G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
    [Crossref]
  13. I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
    [Crossref] [PubMed]
  14. S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
    [Crossref]
  15. D. A. Berthold, G. T. Babcock, and C. A. Yocum, “A highly resolved oxygen-evolving Photosystem II preparation from spinach thylakoid membranes,” FEBS Lett. 134, 231–234 (1981).
    [Crossref]
  16. D. Walker, “Mechanical isolation of chloroplasts” in “The use of oxygen electrodes and fluorescent probes in simple measurements of photosynthesis”, (Oxygraphics, Sheffield, 1987) 107–114.
  17. V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
    [Crossref] [PubMed]
  18. D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
    [Crossref] [PubMed]

2001 (2)

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

2000 (1)

C. Buschmann, G. Langsdorf, and H. K. Lichtenhaler, “Imaging of the blue, green and red fluorescence emission of plants: An overview,” Photosynthetica 38, 483–491 (2000).
[Crossref]

1998 (3)

M. Straub and S. W. Hell, “Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope,” Appl. Phys. Lett. 73, 1769–1771 (1998).
[Crossref]

J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
[Crossref]

K. Dowling, M. J. Dayel, M. J. Lever, P. M. W. French, J. D. Hares, and A. K. L. Dymoke-Bradshaw, “Fluorescence lifetime imaging with picosecond resolution for biomedical applications,” Opt. Lett. 23, 810–812 (1998).
[Crossref]

1997 (1)

J. M. Anderson, Y. -I. Park, and W. S. Chow, “Photoinactivation and photoprotection of Photosystem II in nature”, Physiol. Plantarum 100, 214–223 (1997).
[Crossref]

1996 (2)

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
[Crossref]

1995 (1)

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

1992 (1)

T. A. Roelofs, C. -H. Lee, and A. R. Holzwarth, “Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in Photosystem II alpha- and beta-units,” Biophys. J. 61, 1147–1163 (1992).
[Crossref] [PubMed]

1991 (3)

T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
[Crossref]

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
[Crossref] [PubMed]

1989 (1)

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

1984 (1)

J. F. Bornman, L. O. Bjorn, and H. E. Akerlund, “Action spectrum for inhibition by UV radiation of Photosystem II activity in spinach-oleracea thylakoids,” Photobiochem. & Photobiophys. 8, 305–314 (1984).

1981 (1)

D. A. Berthold, G. T. Babcock, and C. A. Yocum, “A highly resolved oxygen-evolving Photosystem II preparation from spinach thylakoid membranes,” FEBS Lett. 134, 231–234 (1981).
[Crossref]

Akerlund, H. E.

J. F. Bornman, L. O. Bjorn, and H. E. Akerlund, “Action spectrum for inhibition by UV radiation of Photosystem II activity in spinach-oleracea thylakoids,” Photobiochem. & Photobiophys. 8, 305–314 (1984).

Anderson, J. M.

J. M. Anderson, Y. -I. Park, and W. S. Chow, “Photoinactivation and photoprotection of Photosystem II in nature”, Physiol. Plantarum 100, 214–223 (1997).
[Crossref]

Arndt Jovin, D. J.

G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
[Crossref] [PubMed]

Atamian, M.

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

Babcock, G. T.

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

D. A. Berthold, G. T. Babcock, and C. A. Yocum, “A highly resolved oxygen-evolving Photosystem II preparation from spinach thylakoid membranes,” FEBS Lett. 134, 231–234 (1981).
[Crossref]

Bakou, A.

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

Barzda, V.

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

Berthold, D. A.

D. A. Berthold, G. T. Babcock, and C. A. Yocum, “A highly resolved oxygen-evolving Photosystem II preparation from spinach thylakoid membranes,” FEBS Lett. 134, 231–234 (1981).
[Crossref]

Bjorn, L. O.

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

J. F. Bornman, L. O. Bjorn, and H. E. Akerlund, “Action spectrum for inhibition by UV radiation of Photosystem II activity in spinach-oleracea thylakoids,” Photobiochem. & Photobiophys. 8, 305–314 (1984).

Blubaugh, D. J.

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

Bornman, J. F.

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

J. F. Bornman, L. O. Bjorn, and H. E. Akerlund, “Action spectrum for inhibition by UV radiation of Photosystem II activity in spinach-oleracea thylakoids,” Photobiochem. & Photobiophys. 8, 305–314 (1984).

Buschmann, C.

C. Buschmann, G. Langsdorf, and H. K. Lichtenhaler, “Imaging of the blue, green and red fluorescence emission of plants: An overview,” Photosynthetica 38, 483–491 (2000).
[Crossref]

Caldwell, M. M.

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

Cervinskas, V.

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

Cheniae, G. M.

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

Chow, W. S.

J. M. Anderson, Y. -I. Park, and W. S. Chow, “Photoinactivation and photoprotection of Photosystem II in nature”, Physiol. Plantarum 100, 214–223 (1997).
[Crossref]

Clegg, R. M.

G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
[Crossref] [PubMed]

Dayel, M. J.

de Grauw, C. J.

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
[Crossref]

Dowling, K.

Dymoke-Bradshaw, A. K. L.

Eckert, H. -J.

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

French, P. M. W.

Fromme, R.

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

Gerritsen, H. C.

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
[Crossref]

Ghanotakis, D. F.

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

Gilbert, M.

T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
[Crossref]

Goldbeck, J. H.

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

Graeber, P.

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

Gulbinas, V.

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

Hares, J. D.

Hell, S. W.

M. Straub and S. W. Hell, “Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope,” Appl. Phys. Lett. 73, 1769–1771 (1998).
[Crossref]

Hohm-Veit, S.

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

Holzwarth, A. R.

T. A. Roelofs, C. -H. Lee, and A. R. Holzwarth, “Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in Photosystem II alpha- and beta-units,” Biophys. J. 61, 1147–1163 (1992).
[Crossref] [PubMed]

T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
[Crossref]

Jovin, T. M.

G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
[Crossref] [PubMed]

Kananavicius, R.

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

Kleima, F. J.

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

Kulandaivelu, G.

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

Langsdorf, G.

C. Buschmann, G. Langsdorf, and H. K. Lichtenhaler, “Imaging of the blue, green and red fluorescence emission of plants: An overview,” Photosynthetica 38, 483–491 (2000).
[Crossref]

Larkum, A. W. D.

S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
[Crossref]

Lee, C. -H.

T. A. Roelofs, C. -H. Lee, and A. R. Holzwarth, “Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in Photosystem II alpha- and beta-units,” Biophys. J. 61, 1147–1163 (1992).
[Crossref] [PubMed]

Lever, M. J.

Lichtenhaler, H. K.

C. Buschmann, G. Langsdorf, and H. K. Lichtenhaler, “Imaging of the blue, green and red fluorescence emission of plants: An overview,” Photosynthetica 38, 483–491 (2000).
[Crossref]

Lukins, P. B.

S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
[Crossref]

Marriott, G.

G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
[Crossref] [PubMed]

Park, Y. -I.

J. M. Anderson, Y. -I. Park, and W. S. Chow, “Photoinactivation and photoprotection of Photosystem II in nature”, Physiol. Plantarum 100, 214–223 (1997).
[Crossref]

Petrouleas, V.

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

Post, S. A.

S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
[Crossref]

Renger, G.

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

Roelofs, T. A.

T. A. Roelofs, C. -H. Lee, and A. R. Holzwarth, “Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in Photosystem II alpha- and beta-units,” Biophys. J. 61, 1147–1163 (1992).
[Crossref] [PubMed]

T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
[Crossref]

Sass, L.

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

Shuvalov, V. A.

T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
[Crossref]

Spetea, C.

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

Straub, M.

M. Straub and S. W. Hell, “Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope,” Appl. Phys. Lett. 73, 1769–1771 (1998).
[Crossref]

Sytsma, J.

J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
[Crossref]

Teramura, A. H.

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

Tevini, M.

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

Valkunas, L.

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

van Ameroagen, H.

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

van Amerongen, H.

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

van Grondelle, R.

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

Vass, I.

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

Volker, M.

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

Vroom, J. M.

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
[Crossref]

Walker, D.

D. Walker, “Mechanical isolation of chloroplasts” in “The use of oxygen electrodes and fluorescent probes in simple measurements of photosynthesis”, (Oxygraphics, Sheffield, 1987) 107–114.

Walker, P. J.

S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
[Crossref]

Yocum, C. A.

D. A. Berthold, G. T. Babcock, and C. A. Yocum, “A highly resolved oxygen-evolving Photosystem II preparation from spinach thylakoid membranes,” FEBS Lett. 134, 231–234 (1981).
[Crossref]

Ambio (1)

M. M. Caldwell, A. H. Teramura, M. Tevini, J. F. Bornman, L. O. Bjorn, and G. Kulandaivelu, “Effects of increased solar ultraviolet radiation on terrestrial plants”, Ambio 24, 166–173 (1995).

Appl. Phys. Lett. (1)

M. Straub and S. W. Hell, “Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope,” Appl. Phys. Lett. 73, 1769–1771 (1998).
[Crossref]

Biochemistry (2)

I. Vass, L. Sass, C. Spetea, A. Bakou, D. F. Ghanotakis, and V. Petrouleas, “UV-B induced inhibition of Photosystem II electron transport studied by EPR and chlorophyll fluorescence. Impairment of donor- and acceptor-side components,” Biochemistry 35, 8964–8973 (1996).
[Crossref] [PubMed]

D. J. Blubaugh, M. Atamian, G. T. Babcock, J. H. Goldbeck, and G. M. Cheniae, “Photoinhibition of hydroxylamine-extracted Photosystem II membranes: identification of the sites of photodamage,” Biochemistry 30, 7586–7597 (1991).
[Crossref] [PubMed]

Biochim. Biophys. Acta (1)

T. A. Roelofs, M. Gilbert, V. A. Shuvalov, and A. R. Holzwarth, “Picosecond fluorescence kinetics of the D1/D2- cytb559 Photosystem II reaction center complex energy transfer and primary charge separation processes,” Biochim. Biophys. Acta 1060, 237–244 (1991).
[Crossref]

Biophys. J. (4)

T. A. Roelofs, C. -H. Lee, and A. R. Holzwarth, “Global target analysis of picosecond chlorophyll fluorescence kinetics from pea chloroplasts. A new approach to the characterization of the primary processes in Photosystem II alpha- and beta-units,” Biophys. J. 61, 1147–1163 (1992).
[Crossref] [PubMed]

V. Barzda, V. Gulbinas, R. Kananavicius, V. Cervinskas, H. van Ameroagen, R. van Grondelle, and L. Valkunas, “Singlet-singlet annihilation kinetics in aggregates and trimers of LHC II,” Biophys. J. 80, 2409–2421 (2001).
[Crossref] [PubMed]

G. Marriott, R. M. Clegg, D. J. Arndt Jovin, and T. M. Jovin, “Time-resolved imaging microscopy: fluorescence and delayed fluorescence imaging,” Biophys. J. 60, 1374–1387 (1991).
[Crossref] [PubMed]

V. Barzda, C. J. de Grauw, J. M. Vroom, F. J. Kleima, R. van Grondelle, H. van Amerongen, and H. C. Gerritsen, “Fluorescence lifetime heterogeneity in aggregates of LHC II revealed by time-resolved microscopy,” Biophys. J. 81, 538–546 (2001).
[Crossref] [PubMed]

FEBS Lett. (1)

D. A. Berthold, G. T. Babcock, and C. A. Yocum, “A highly resolved oxygen-evolving Photosystem II preparation from spinach thylakoid membranes,” FEBS Lett. 134, 231–234 (1981).
[Crossref]

J. Microsc. (1)

J. Sytsma, J. M. Vroom, C. J. de Grauw, and H. C. Gerritsen, “Time-gated fluorescence lifetime imaging and microvolume spectroscopy using two-photon excitation,” J. Microsc. 191, 39–51 (1998)
[Crossref]

Opt. Lett. (1)

Photobiochem. & Photobiophys. (1)

J. F. Bornman, L. O. Bjorn, and H. E. Akerlund, “Action spectrum for inhibition by UV radiation of Photosystem II activity in spinach-oleracea thylakoids,” Photobiochem. & Photobiophys. 8, 305–314 (1984).

Photochem. Photobiol. (1)

G. Renger, M. Volker, H. -J. Eckert, R. Fromme, S. Hohm-Veit, and P. Graeber, “On the mechanism of Photosystem II deterioration by UV-B irradiation,” Photochem. Photobiol. 49, 97–106 (1989).
[Crossref]

Photosynth. Res. (1)

S. A. Post, P. B. Lukins, P. J. Walker, and A. W. D. Larkum, “The effects of ultraviolet radiation on P680+ reduction in PS II core complexes measured in individual S-states and during repetitive cycling of the oxygen-evolving complex,” Photosynth. Res. 49, 21–27 (1996).
[Crossref]

Photosynthetica (1)

C. Buschmann, G. Langsdorf, and H. K. Lichtenhaler, “Imaging of the blue, green and red fluorescence emission of plants: An overview,” Photosynthetica 38, 483–491 (2000).
[Crossref]

Physiol. Plantarum (1)

J. M. Anderson, Y. -I. Park, and W. S. Chow, “Photoinactivation and photoprotection of Photosystem II in nature”, Physiol. Plantarum 100, 214–223 (1997).
[Crossref]

Other (1)

D. Walker, “Mechanical isolation of chloroplasts” in “The use of oxygen electrodes and fluorescent probes in simple measurements of photosynthesis”, (Oxygraphics, Sheffield, 1987) 107–114.

Supplementary Material (1)

» Media 1: MOV (1268 KB)     

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Figures (4)

Fig. 1.
Fig. 1.

Schematic diagram of picosecond time-gated fluorescence microscope.

Fig. 2.
Fig. 2.

Fluorescence images for the PS II membrane fragments (open centers) and fluorescence decay curves for various chloroplast and PS II preparations. The inset in the image shows a 2x zoom of a region in which aggregation occurs.

Fig. 3.
Fig. 3.

Picosecond time-resolved imaging of chlorophyll fluorescence from untreated (left) and 300 nm UV-irradiated (right) whole spinach leaf tissue (100 ps per s). Animation.mov (1.3 Mb).

Fig. 4.
Fig. 4.

Fluorescence decay curves for untreated and UV-irradiated whole spinach leaf tissue.

Tables (1)

Tables Icon

Table 1. Fluorescence decay amplitudes and time constants for photosynthetic systemsa.

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