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Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy.

Publication ,  Journal Article
Trivedi, NS; Wang, HW; Nieminen, AL; Oleinick, NL; Izatt, JA
Published in: Photochemistry and photobiology
May 2000

Photodynamic therapy (PDT) is a novel cancer therapy that uses light-activated drugs (photosensitizers) to destroy tumor tissue. Reactive oxygen species produced during PDT are thought to cause the destruction of tumor tissue. However, the precise mechanism of PDT is not completely understood. To provide insight into the in vitro mechanisms of PDT, we studied the subcellular localization of the photosensitizer HOSiPcOSi(CH3)2-(CH2)3N(CH3)2 (Pc 4) in mouse lymphoma (LY-R) cells using double-label confocal fluorescence microscopy. This technique allowed us to observe the relative distributions of Pc 4 and an organelle-specific dye within the same cell via two, spectrally distinct, fluorescence images. To quantify the localization of Pc 4 within different organelles, linear correlation coefficients from the fluorescence data of Pc 4 and the organelle-specific dyes were calculated. Using this measurement, the subcellular spatial distributions of Pc 4 could be successfully monitored over an 18 h period. At early times (0-1 h) after introduction of Pc 4 to LY-R cells, the dye was found in the mitochondria, lysosomes and Golgi apparatus, as well as other cytoplasmic membranes, but not in the plasma membrane or the nucleus. Over the next 2 h, there was some loss of Pc 4 from the lysosomes as shown by the correlation coefficients. After an additional incubation period of 2 h Pc 4 slowly increased its accumulation in the lysosomes. The highest correlation coefficient (0.65) was for Pc 4 and BODIPY-FL C5 ceramide, which targets the Golgi apparatus, and also binds to other cytoplasmic membranes. The correlation coefficient was also high (0.60) for Pc 4 and a mitochondria-targeting dye (Mitotracker Green FM). Both of these correlation coefficients were higher than that for Pc 4 with the lysosome-targeting dye (Lysotracker Green DND-26). The results suggest that Pc 4 binds preferentially and strongly to mitochondria and Golgi complexes.

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Published In

Photochemistry and photobiology

DOI

EISSN

1751-1097

ISSN

0031-8655

Publication Date

May 2000

Volume

71

Issue

5

Start / End Page

634 / 639

Related Subject Headings

  • Tumor Cells, Cultured
  • Photosensitizing Agents
  • Photochemotherapy
  • Microscopy, Fluorescence
  • Microscopy, Confocal
  • Mice
  • Lymphoma
  • Indoles
  • Biophysics
  • Animals
 

Citation

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ICMJE
MLA
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Trivedi, N. S., Wang, H. W., Nieminen, A. L., Oleinick, N. L., & Izatt, J. A. (2000). Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy. Photochemistry and Photobiology, 71(5), 634–639. https://doi.org/10.1562/0031-8655(2000)071<0634:qaopli>2.0.co;2
Trivedi, N. S., H. W. Wang, A. L. Nieminen, N. L. Oleinick, and J. A. Izatt. “Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy.Photochemistry and Photobiology 71, no. 5 (May 2000): 634–39. https://doi.org/10.1562/0031-8655(2000)071<0634:qaopli>2.0.co;2.
Trivedi NS, Wang HW, Nieminen AL, Oleinick NL, Izatt JA. Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy. Photochemistry and photobiology. 2000 May;71(5):634–9.
Trivedi, N. S., et al. “Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy.Photochemistry and Photobiology, vol. 71, no. 5, May 2000, pp. 634–39. Epmc, doi:10.1562/0031-8655(2000)071<0634:qaopli>2.0.co;2.
Trivedi NS, Wang HW, Nieminen AL, Oleinick NL, Izatt JA. Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy. Photochemistry and photobiology. 2000 May;71(5):634–639.
Journal cover image

Published In

Photochemistry and photobiology

DOI

EISSN

1751-1097

ISSN

0031-8655

Publication Date

May 2000

Volume

71

Issue

5

Start / End Page

634 / 639

Related Subject Headings

  • Tumor Cells, Cultured
  • Photosensitizing Agents
  • Photochemotherapy
  • Microscopy, Fluorescence
  • Microscopy, Confocal
  • Mice
  • Lymphoma
  • Indoles
  • Biophysics
  • Animals