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Second harmonic imaging of chloroplasts using the two-photon laser scanning microscope.

Ali Hussain Reshak¹, Vassilios Sarafis, Rainer Heintzmann

¹Institute of Physical Biology, University of South Bohemia, Nove Hrady 37333, Czech Republic. maalidph@yahoo.co.uk

Micron (Oxford, England : 1993)

|November 26, 2008

Summary

Second harmonic generation (SHG) imaging successfully visualized chloroplast grana in moss leaves. This technique revealed grana structures and their reorientation under illumination, offering insights into plant cell organization.

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Area of Science:

Plant Biology
Cellular Imaging
Biophysics

Background:

Second harmonic generation (SHG) provides insights into asymmetric, spatially oriented structures.
Chloroplasts, the sites of photosynthesis, possess internal structures like grana.

Purpose of the Study:

To investigate the structure of chloroplast grana and intergranal regions in the moss *Plagiomnium affine* using SHG imaging.
To assess the feasibility of SHG for visualizing chloroplast substructures without interference from starch granules.

Main Methods:

Utilized a two-photon microscope equipped with blocking filters to capture SHG signals.
Employed SHG imaging on chloroplasts from *Plagiomnium affine* leaf cells, specifically targeting starch-free regions.

Main Results:

Strong SHG signals were detected specifically from the granal regions of chloroplasts.
Observed that chloroplasts reoriented their orientation upon illumination, influencing the SHG signal intensity.
Successfully obtained SHG signals from grana, free from confounding signals from starch grains.

Conclusions:

SHG imaging is a viable method for visualizing chloroplast grana in moss leaves.
The study demonstrates the potential of SHG to reveal structural details and dynamic reorientation of chloroplasts.