Serial Block Face Imaging

Abstract

We have recently installed a serial block face imaging system and have collected our first major data set. The project was a proof of concept to track the progress of pollen tubes towards specialist cells involved in plant reproduction. Pollen tubes target synergid cells and in particular the basal membrane of those cells which is known as the filiform apparatus. Little to nothing in known about the behavior of the filiform apparatus during pollen tube reception. By examining the ovules of Arabidopsis in 3D volumes we hope to examine whether the pollen tube enters the receptive synergid through the filiform apparatus, In addition we hope to determine if the pollen tube enters the receptive synergid at all. We prepared samples using an enhanced protocol that has been designed to improve signal and contrast in samples for serial block face imaging. We collected close to 1800 40nm slices from 3 different regions. We successfully found and oriented the two synergids in different ovules. We used Amira for visualization and have carried out segmentation of the filiform apparatus.

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