Abstract
A recently introduced helium ion microscopy (HIM) was employed to observe uncoated pine leaf specimens. Adult leaves were collected from the seedlings of Pinus densiflora and P. rigida, air-dried at room temperature, and observed by HIM without metal coating. Ovoid or round stomata and distinct Florin rings could be discerned. The epicuticular waxes were present in the epistomatal chambers and Florin rings of stomata on the leaf surface. The epicuticular waxes were mostly straight, cylindrical, and ca. 1 <TEX>${\mu}m$</TEX> in length. The epistomatal chambers of P. rigida were filled with the epicuticular waxes, whereas those of P. densiflora were not filled with the epicuticular waxes. Based on their micromorphology, the epicuticular wax structures of the pine species were identified as tubules. These results suggest that the HIM could be used for the investigation of the plant stomata and epicuticular waxes of uncoated plant leaves. Due to the smaller ion probe and interaction volume, the HIM has advantages over conventional field emission scanning electron microscopy in terms of image resolution and charge neutralization.
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