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Imaging granularity of leukocytes with third harmonic generation microscopy.

Cheng-Kun Tsai1, Yu-Shing Chen, Pei-Chun Wu

  • 1Institute of Biomedical Engineering, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei 10617, Taiwan.

Biomedical Optics Express
|October 2, 2012
PubMed
Summary

Label-free third harmonic generation (THG) microscopy differentiates leukocyte granularity. This technique allows in vivo tracking of leukocyte recruitment, offering a non-invasive method for studying cellular responses.

Keywords:
(170.1530) Cell analysis(180.4315) Nonlinear microscopy

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Area of Science:

  • Biomedical Optics
  • Cellular Biology
  • Immunology

Background:

  • Leukocyte identification and tracking are crucial in understanding immune responses.
  • Current methods often require cell labeling, which can alter cellular function.
  • Developing label-free techniques is essential for non-invasive in vivo studies.

Purpose of the Study:

  • To investigate the potential of third harmonic generation (THG) microscopy for label-free differentiation of leukocyte subtypes based on granularity.
  • To assess the feasibility of using THG microscopy for in vivo tracking of leukocyte recruitment.

Main Methods:

  • Utilizing a 1230 nm femtosecond laser for THG excitation.
  • Employing THG microscopy to image leukocytes.
  • Observing leukocyte recruitment in vivo following lipopolysaccharide (LPS) challenge.

Main Results:

  • THG signals were significantly higher in neutrophils compared to other mononuclear cells.
  • Agranular lymphocytes exhibited THG signals one order of magnitude smaller than neutrophils.
  • Characteristic THG features were observed in vivo, enabling tracking of newly recruited leukocytes.

Conclusions:

  • Label-free THG microscopy can effectively distinguish leukocyte subtypes based on granularity.
  • In vivo THG imaging allows for timely tracking of leukocyte movement without perturbing normal cellular or physiological states.
  • THG microscopy presents a promising tool for non-invasive monitoring of immune cell dynamics.