May 26, 2023

Public workspaceAncient dental calculus sampling procedure for optical microscopic analysis

DOI
https://dx.doi.org/10.17504/protocols.io.q26g7yqpkgwz/v1
Ancient dental calculus sampling procedure for optical microscopic analysis
  • Dulce Neves1,2,
  • Emanuela Cristiani3,
  • António austino Carvalho4,
  • Ana Maria Silva1,5,6
  • 1University of Coimbra, Research Centre for Anthropology and Health, Department of Life Sciences, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal;
  • 2Sapienza University of Rome, Department of History, Anthropology, Religion, and Performing Arts, Piazzale Aldo Moro, 00185 Rome, Italy;
  • 3DANTE – Diet and Ancient Technology Laboratory, Department of Oral and Maxillo-Facial Sciences,Sapienza Università di Roma, Roma, Italy;
  • 4CEAACP - Centro de Estudos de Arqueologia, Artes e Ciências do Património, University of Algarve, FCHS, Campus de Gambelas, 8000-117, Faro, Portugal;
  • 5UNIARQ – University of Lisbon, Portugal;
  • 6University of Coimbra, Centre for Functional Ecology, Department of Life Sciences, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal
Dulce Neves
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DOI: https://dx.doi.org/10.17504/protocols.io.q26g7yqpkgwz/v1
Protocol Citation: Dulce Neves, Emanuela Cristiani, António austino Carvalho, Ana Maria Silva 2023. Ancient dental calculus sampling procedure for optical microscopic analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.q26g7yqpkgwz/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 26, 2023
Last Modified: May 26, 2023
Protocol Integer ID: 82531
Keywords: dental calculus, sampling, archaeobotany, archaeology, bioarchaeology, bioanthropology, teeth, dental remains, dental anthropology, skeleton, light microscopy, ancient dental calculus, dental calculus from individual teeth, procedure for optical microscopic analysis, matrix of the dental calculus, optical microscopic analysis, optical microscopy analysis, individual teeth, biomolecular analysis, sampling procedure, proteomic, sample
Funders Acknowledgements:
FCT - Fundação para a Ciência e a Tecnologia (PhD Studentship)
Grant ID: 2021.04560.BD
FCT - Fundação para a Ciência e Tecnologia
Grant ID: UIDB/00283/2020
ERC Starting Grant Project
Grant ID: 639286
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).
Guidelines
A clean sterile sampling environment is a vital factor to reduce contamination.

Gloves should be worn during the whole sampling procedure.
Materials
  • Powder-free nitrile gloves
  • Aluminum foil
  • Plastic wrap
  • Alcohol
  • Paper towels
  • Sterile disposable blades
  • Centrifuge tubes (1.5 or 2 ml)
  • Centrifuge tube storage and transport box
  • Precision scale
  • Vernier caliper
  • Magnifying lens (tube/bag labeling)
  • Sharpie
  • Camera and scale
  • Metadata sheet (including Sampling ID; Collection ID; Burial/Tooth ID; Tooth FDI Code; Surface; Pathologies; Other Notes)
Troubleshooting
Safety warnings
Be careful when handling the disposable blade.
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)

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
LINK

Citation
B. Holly Smith (1983). Patterns of molar wear in hunter–gatherers and agriculturalists. Am. J. Phys. Anthropol., 63: 39-56.
https://doi.org/10.1002/ajpa.1330630107
LINK

Citation
Jane E. Buikstra and Douglas H. Ubelaker (1993). 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
LINK


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

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

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
Citations
Step 5
B. Holly Smith. Patterns of molar wear in hunter–gatherers and agriculturalists
https://doi.org/10.1002/ajpa.1330630107
Step 5
Jane E. Buikstra and Douglas H. Ubelaker. 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
Step 5
Simon Hillson. Recording dental caries in archaeological human remains
https://doi.org/10.1002/oa.538