Ancient dental calculus sampling procedure for optical microscopic analysis

Dulce Neves, Emanuela Cristiani, António Faustino Carvalho, Ana Maria Silva

Published: 2023-05-27 DOI: 10.17504/protocols.io.q26g7yqpkgwz/v1

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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

1.

Clean the working table surface using alcohol and paper towels, and use powder-free nitrile gloves throughout the whole steps

2.

Make sure to sterilize all utensils used before starting the sample collection and between sampling each tooth. Use alcohol and the paper towels

3.

Cover the surface with plastic wrap

4.

Label the centrifuge tube ready for sampling

5.

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)

Note

For tooth numbering, use the FDI system.

Citation

Simon Hillson 2001 Recording dental caries in archaeological human remains Int. J. Osteoarchaeol., 11: 249-289 https://doi.org/10.1002/oa.538

Citation

B. Holly Smith 1984 Patterns of molar wear in hunter–gatherers and agriculturalists Am. J. Phys. Anthropol., 63: 39-56 https://doi.org/10.1002/ajpa.1330630107

Citation

Jane E. Buikstra and Douglas H. Ubelaker 1994 Standards for Data Collection from Human Skeletal Remains: Proceedings of a Seminar at the Field Museum of Natural History organized by Jonathan Haas ISBN 1-56349-075-7

6.

Measure the dental calculus deposits using the Vernier caliper

7.

Photograph the tooth surfaces before collecting the dental calculus deposit

8.

Display aluminum foil on the working table and carefully collect the dental calculus deposit using a sterile disposable blade and under the magnifying lens

Note

Be careful not to scratch the enamel/tooth surface with the tip of the disposable blade.

9.

Put the sample in the labelled centrifuge tube

10.

Remove and discard the piece of foil

11.

Weight the deposit using a precision scale

12.

Take new photos of the tooth after the collection of the dental calculus deposit

Control samples

13.

Take control samples of the working table surface (and other relevant surfaces) and maxillary bone (if present) to cross-out possible contamination

Ancient dental calculus sampling procedure for optical microscopic analysis

Disclaimer

Abstract

Steps

Sampling preparation and dental calculus collection

Control samples

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