Jan 29, 2025

Public workspaceAn effective protocol to isolate and mechanically test silk fibers spun by Osmia lignaria

Peer-reviewed method
  • 1Department of Biology, Utah State University, Logan, Utah, USA;
  • 2Department of Chemistry and Biochemistry, Utah State University, Logan, Utah, USA;
  • 3Pollinating Insect Research Unit, Pacific West Area, Agricultural Research Service, United States Department of Agriculture, Logan, Utah, USA
  • Oran Wasserman: ‡Authors contributed equally;
  • Jackson J. Morley: ‡Authors contributed equally;
  • Mary-Kate F. Williams: ‡Authors contributed equally
  • PLOS ONE Lab Protocols
    Tech. support email: plosone@plos.org
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Collection CitationOran Wasserman, Jackson J. Morley, Mary-Kate F. Williams, Brianne E. Bell, Diana L Cox-Foster, Justin A. Jones 2025. An effective protocol to isolate and mechanically test silk fibers spun by Osmia lignaria. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6d8bkvqe/v1
Manuscript citation:
Wasserman O, Morley JJ, Williams MF, Bell BE, Cox-Foster DL, Jones JA (2025) An effective protocol to isolate and mechanically test silk fibers spun by Osmia lignaria Say (Hymenoptera: Megachilidae) fifth instar larvae. PLOS One 20(2). doi: 10.1371/journal.pone.0318918
License: This is an open access collection 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 collection and it's working
Created: December 12, 2024
Last Modified: January 29, 2025
Collection Integer ID: 115848
Keywords: silk, isolation, solitary, bee, mechanical, testing
Abstract
Silk, a remarkable protein-based fiber spun by various arthropod lineages, has been prized for millennia, with the cocoon silk of domesticated silkworms and spiders being the most utilized and extensively studied. There is limited information on how silk can be used to investigate biology, development, and health in other silk producing species, particularly for solitary bees such as Osmia lignaria Say (Hymenoptera: Megachilidae). Osmia lignaria, an increasingly managed solitary pollinator, produces silk cocoons during the fifth instar larval stage. We have developed a minimally invasive protocol to isolate and mechanically test O. lignaria silk fibers using a 3-D printed well plate system for rearing and two specific isolation techniques. Our protocol allows for collecting individual fibers directly from silk-spinning larvae between silk initiation and cocoon formation without preventing subsequent cocoon development, enabling silk characterization as part of larger rearing and developmental studies. For this study, isolated fibers were mounted on C-cards, facilitating diameter measurement using a microscope and mechanical testing with an MTS Synergie 100 tensile testing instrument. We successfully isolated and tested the mechanical properties of naturally spun silk from O. lignaria, with 20 fibers isolated and mechanically tested from seven larvae. Further examination of isolated silk can reveal physical, molecular, chemical, and morphological characteristics, advancing our understanding of bee silk properties and their role in bee biology, evolution, and nutritional status. This protocol provides a practical tool for researchers to isolate and study silk from silk-producing bee species.
Files
Protocol
Icon representing the file C-cards preparations
Name
C-cards preparations
Version 1
, USU
Oran WassermanUSU
Protocol
Icon representing the file Fiber Isolation and Mounting onto C-cards
Name
Fiber Isolation and Mounting onto C-cards
Version 1
, USU
Oran WassermanUSU
Protocol
Icon representing the file Fiber Diameter Measurement and Mechanical Testing
Name
Fiber Diameter Measurement and Mechanical Testing
Version 1
, USU
Oran WassermanUSU