Light-induced cell damage in live-cell super-resolution microscopy

Sina Wäldchen, Julian Lehmann, Teresa Klein, Sebastian van de Linde, Markus Sauer

Research output: Contribution to journalArticle

152 Citations (Scopus)

Abstract

Super-resolution microscopy can unravel previously hidden details of cellular structures but requires high irradiation intensities to use the limited photon budget efficiently. Such high photon densities are likely to induce cellular damage in live-cell experiments. We applied single-molecule localization microscopy conditions and tested the influence of irradiation intensity, illumination-mode, wavelength, light-dose, temperature and fluorescence labeling on the survival probability of different cell lines 20-24 hours after irradiation. In addition, we measured the microtubule growth speed after irradiation. The photo-sensitivity is dramatically increased at lower irradiation wavelength. We observed fixation, plasma membrane permeabilization and cytoskeleton destruction upon irradiation with shorter wavelengths. While cells stand light intensities of ~1 kW cm(-2) at 640 nm for several minutes, the maximum dose at 405 nm is only ~50 J cm(-2), emphasizing red fluorophores for live-cell localization microscopy. We also present strategies to minimize phototoxic factors and maximize the cells ability to cope with higher irradiation intensities.
Original languageEnglish
Article number15348
Number of pages12
JournalScientific Reports
Volume5
DOIs
Publication statusPublished - 20 Oct 2015
Externally publishedYes

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damage
microscopy
irradiation
cells
wavelengths
dosage
photon density
photosensitivity
cultured cells
budgets
luminous intensity
marking
destruction
illumination
membranes
fluorescence
photons
molecules
temperature

Keywords

  • single-molecule localization microscopy
  • cell damage

Cite this

Wäldchen, Sina ; Lehmann, Julian ; Klein, Teresa ; van de Linde, Sebastian ; Sauer, Markus. / Light-induced cell damage in live-cell super-resolution microscopy. In: Scientific Reports. 2015 ; Vol. 5.
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Light-induced cell damage in live-cell super-resolution microscopy. / Wäldchen, Sina; Lehmann, Julian; Klein, Teresa; van de Linde, Sebastian; Sauer, Markus.

In: Scientific Reports, Vol. 5, 15348, 20.10.2015.

Research output: Contribution to journalArticle

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AU - Wäldchen, Sina

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AB - Super-resolution microscopy can unravel previously hidden details of cellular structures but requires high irradiation intensities to use the limited photon budget efficiently. Such high photon densities are likely to induce cellular damage in live-cell experiments. We applied single-molecule localization microscopy conditions and tested the influence of irradiation intensity, illumination-mode, wavelength, light-dose, temperature and fluorescence labeling on the survival probability of different cell lines 20-24 hours after irradiation. In addition, we measured the microtubule growth speed after irradiation. The photo-sensitivity is dramatically increased at lower irradiation wavelength. We observed fixation, plasma membrane permeabilization and cytoskeleton destruction upon irradiation with shorter wavelengths. While cells stand light intensities of ~1 kW cm(-2) at 640 nm for several minutes, the maximum dose at 405 nm is only ~50 J cm(-2), emphasizing red fluorophores for live-cell localization microscopy. We also present strategies to minimize phototoxic factors and maximize the cells ability to cope with higher irradiation intensities.

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