Abstract
An advanced protocol to prepare single extant and fossil pollen grains for transmission electron microscopy (TEM) analysis allows for the fast recovery of data on the ultrastructure of pollen/spores. The protocol is easy to apply and less time consuming than previous methods. The ‘loss’ of pollen grains and pollen that is ‘difficult to locate’ within the embedding material is avoided, and each single pollen grain can be prepared successfully for TEM analysis. This preparation method is meant as an addition to the single-grain method using combined light and scanning electron microscopy to investigate dispersed fossil pollen grains developed by Dr Reinhard Zetter in the late 1980s.
Highlights
An advanced protocol to prepare single extant and fossil pollen grains for transmission electron microscopy (TEM) analysis allows for the fast recovery of data on the ultrastructure of pollen/spores
The ‘loss’ of pollen grains and pollen that is ‘difficult to locate’ within the embedding material is avoided, and each single pollen grain can be prepared successfully for TEM analysis. This preparation method is meant as an addition to the single-grain method using combined light and scanning electron microscopy to investigate dispersed fossil pollen grains developed by Dr Reinhard Zetter in the late 1980s
Prefixation with glutaraldehyde is important to preserve the structure of the living protoplast in pollen, but since fossil pollen grains are without cell content there is no need for this preparation step
Summary
An advanced protocol to prepare single extant and fossil pollen grains for transmission electron microscopy (TEM) analysis allows for the fast recovery of data on the ultrastructure of pollen/spores. In most palaeobotanical literature, such a comparison is based on morphological features that can be observed using light microscopy (LM) only This is despite the fact that over the past 50 years papers documenting morphology and ultrastructure in pollen of extant plants have accumulated drastically. Following SEM observations, the grains were transported and prepared for TEM according to the procedure presented by Doyle et al (1975). The grains had only to be transported twice, and without any effort the pollen would attach to the nasal hair when soaked in glycerine This method has been used in numerous papers by Reinhard Zetter and his colleagues since its introduction 30 years ago It is difficult to find any good reason for this lack of TEM analysis, but we speculate that the time consuming
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