Shallow-layer microbe harvest and data display method shows taxa pre-harvest spatial positions

We report a new harvest method and data display of taxa in material dissected from within dental caries lesions for culture or non-culture sequencing and taxa identification. This method is significant because it preserves the microbe spatial positions relative to lesion depth and uses micro-surgical aseptic techniques that allow exact dissection of precisely selected portions of the lesion while preventing contamination. Samples are harvested in shallow layers and each layer harvested becomes a separate sample for sequencing and taxa identification. The layers are coded in a way that allows sequential reassembling of the layers after taxa identification. Reassembling the harvested layers is analogous to reassembling pieces in a puzzle that rebuild a picture. In this case, the reassembled layers build a "lesion diagram" showing where within the pre-harvest lesion the identified taxa resided relative to lesion depth in the pulpal direction. After reassembling lesions, taxa detail not possible previously showed taxa location, abundance and diversity relative to lesion pulpal depth, plus absolute abundance per milligram and taxa transitions and fluctuations from superficial through deepest dissected layers.