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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Updated: Jun 26, 2026

A Rapid Laser Probing Method Facilitates the Non-invasive and Contact-free Determination of Leaf Thermal Properties
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Second harmonic generation from thick leaves using the two-photon laser scanning microscope.

Ali Hussain Reshak1

  • 1Institute of Physical Biology-South Bohemia University, Institute of System Biology and Ecology-Academy of Sciences, Nove Hrady, Czech Republic. maalidph@yahoo.co.uk

Micron (Oxford, England : 1993)
|February 4, 2009
PubMed
Summary
This summary is machine-generated.

Second harmonic generation (SHG) microscopy provides high-resolution, label-free imaging of biological structures like duckweed leaves. This technique avoids photobleaching and photodamage, making it ideal for sensitive or unknown samples.

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Area of Science:

  • Biophysics
  • Microscopy
  • Plant Biology

Background:

  • Traditional imaging techniques can damage sensitive biological samples.
  • Investigating unknown samples often requires lengthy exposure to laser radiation, leading to photobleaching and photodamage.
  • Label-free imaging is desirable for preserving sample integrity and studying intrinsic structures.

Purpose of the Study:

  • To demonstrate the utility of second harmonic generation (SHG) microscopy for high-resolution imaging of biological samples.
  • To investigate the dependence of SHG signal on sample thickness, orientation, and illumination polarization.
  • To establish SHG as a non-invasive imaging modality for sensitive and unknown biological specimens.

Main Methods:

  • Utilized a laser-scanning microscope employing second harmonic generation (SHG) as a probe.
  • Generated SHG by focusing a pulsed laser beam onto multi-cell layer thick duckweed (Lemna minuta) leaves.
  • Collected forward and backward SHG digital images using two objectives.

Main Results:

  • Achieved high-resolution imaging of duckweed leaves.
  • Observed that SHG signal intensity in forward and backward directions correlates with chloroplast thickness.
  • Demonstrated that image intensity is dependent on chloroplast orientation relative to illumination polarization, with light-induced re-orientation observable.

Conclusions:

  • SHG microscopy offers a novel, compact, and non-invasive technique for investigating the true architecture of sensitive, unknown, or non-fluorescent samples.
  • SHG imaging avoids photobleaching and photodamage, preserving sample integrity for subsequent analysis.
  • The technique enables label-free imaging of highly ordered structural proteins under physiological conditions, eliminating the need for sample slicing or exogenous labels.