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Related Experiment Videos

Autofluorescence spectroscopy of optically trapped cells

K König1, Y Liu, G J Sonek

  • 1Beckman Laser Institute and Medical Clinic, University of California, Irvine 92715, USA.

Photochemistry and Photobiology
|November 1, 1995
PubMed
Summary

Near-infrared (NIR) optical tweezers at 1064 nm do not alter cellular autofluorescence, indicating no change in redox state. However, 760 nm NIR and UVA light can induce oxidative stress via autofluorescence changes.

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Area of Science:

  • Biophysics
  • Cellular Physiology
  • Spectroscopy

Background:

  • Cellular autofluorescence provides insights into metabolic states, particularly the redox balance of NAD(P)H.
  • Optical tweezers are used to manipulate cells and probe their physiological responses to light.
  • Understanding the effects of different light wavelengths on cellular autofluorescence is crucial for interpreting experimental results.

Purpose of the Study:

  • To investigate the physiological impact of near-infrared (NIR) and UVA microirradiation on cellular autofluorescence.
  • To determine if 1064 nm NIR optical trapping alters cellular autofluorescence and redox state.
  • To compare the effects of 760 nm NIR and UVA light with 1064 nm NIR light on cellular autofluorescence.

Main Methods:

  • Monitoring cellular autofluorescence spectra using a single-beam gradient force optical trap (optical tweezers).

Related Experiment Videos

  • Exposing Chinese hamster ovary (CHO) cells to 1064 nm NIR, 760 nm NIR, and 365 nm UVA microirradiation.
  • Analyzing changes in autofluorescence intensity and spectral characteristics under different irradiation conditions.

    • Main Results:

    • 1064 nm NIR microbeams did not significantly alter autofluorescence intensity or spectral characteristics, even at high power densities.
    • A 760 nm trapping beam caused a two-fold increase in autofluorescence within 5 minutes.
    • Exposure to 365 nm UVA light during 1064 nm trapping resulted in a five-fold autofluorescence increase and a 6 nm red shift within 10 minutes.

    Conclusions:

    • 1064 nm microbeams are suitable for extended cellular manipulation without inducing autofluorescence changes indicative of altered redox state.
    • 760 nm NIR light may induce cellular stress through a two-photon absorption mechanism, similar to UVA exposure.
    • Both 760 nm NIR and UVA irradiation can alter cellular redox balance, potentially leading to oxidative stress.