Ancient dental calculus sampling procedure for optical microscopic analysis
Dulce Neves, Emanuela Cristiani, António Faustino Carvalho, Ana Maria Silva
dental calculus
sampling
archaeobotany
archaeology
bioarchaeology
bioanthropology
teeth
dental remains
dental anthropology
skeleton
light microscopy
Disclaimer
Abstract
This protocol describes how to sample dental calculus from individual teeth for optical microscopy analysis. The primary use-case is for the analysis of dental micro-remains embedded in the matrix of the dental calculus (e.g. starches, phytoliths, pollens, fungi, fibers, etc.), though it can also be used for biomolecular analysis (DNA and proteomics).
Steps
Sampling preparation and dental calculus collection
Clean the working table surface using alcohol and paper towels, and use powder-free nitrile gloves throughout the whole steps
Make sure to sterilize all utensils used before starting the sample collection and between sampling each tooth. Use alcohol and the paper towels
Cover the surface with plastic wrap
Label the centrifuge tube ready for sampling
Fill in the metadata sheet with all the information and the corresponding tubes IDs. Record the dental pathologies (Hillson, 2001), dental wear (Smith, 1984) and describe the deposit of dental calculus accordingly to its type and size (Buikstra and Ubelaker, 1994)
Measure the dental calculus deposits using the Vernier caliper
Photograph the tooth surfaces before collecting the dental calculus deposit
Display aluminum foil on the working table and carefully collect the dental calculus deposit using a sterile disposable blade and under the magnifying lens
Put the sample in the labelled centrifuge tube
Remove and discard the piece of foil
Weight the deposit using a precision scale
Take new photos of the tooth after the collection of the dental calculus deposit
Control samples
Take control samples of the working table surface (and other relevant surfaces) and maxillary bone (if present) to cross-out possible contamination
