DOI QR코드

DOI QR Code

Application of Autofluorescence for Confocal Microscopy to Aid in Archaeoparasitological Analyses

  • Morrow, Johnica Jo (Pre-Health Pathways, Student Success Center, South Dakota School of Mines & Technology) ;
  • Elowsky, Christian (Department of Agronomy and Horticulture, University of Nebraska-Lincoln)
  • Received : 2019.07.31
  • Accepted : 2019.10.12
  • Published : 2019.12.31

Abstract

Confocal laser scanning microscopy (CLSM) was used to examine archaeoparasitological specimens from coprolites associated with La Cueva de los Muertos Chiquitos (CMC) located near present-day Durango, Mexico. The eggs for 4 different types of parasites recovered from CMC coprolites were imaged using CLSM to assist with identification efforts. While some of the parasite eggs recovered from CMC coprolites were readily identified using standard light microscopy (LM), CLSM provided useful data for more challenging identifications by highlighting subtle morphological features and enhancing visualization of parasite egg anatomy. While other advanced microscopy techniques, such as scanning electron microscopy (SEM), may also detect cryptic identifying characters, CLSM is less destructive to the specimens. Utilizing CLSM allows for subsequent examinations, such as molecular analyses, that cannot be performed following SEM sample preparation and imaging. Furthermore, CLSM detects intrinsic autofluorescence molecules, making improved identification independent of resource and time-intensive protocols. These aspects of CLSM make it an excellent method for assisting in taxonomic identification and for acquiring more detailed images of archaeoparasitological specimens.

Keywords

References

  1. Bohm A, Bartel D, Szucsich NU, Pass G. Confocal imaging of the exo- and endoskeleton of Protura after nondestructive DNA extraction. Soil Organisms 2011; 83: 335-345.
  2. Chetverikoc PE. Confocal laser scanning microscopy technique for the study of internal genitalia and external morphology of eriophyoid mites (Acari: Eriophyoidea). Zootaxa 2012; 3453: 56-68. https://doi.org/10.11646/zootaxa.3453.1.4
  3. Grzywacz A, Goral T, Szpila K, Hall MJ. Confocal laser scanning microscopy as a valuable tool in Diptera larval morphology studies. Parasitol Res 2014; 113: 4297-4302. https://doi.org/10.1007/s00436-014-4125-0
  4. Klaus AV, Schawaroch V. Novel methodology utilizing confocal laser scanning microscopy for systematic analysis in arthropods (Insecta). Integr Comp Biol 2006; 46: 207-214. https://doi.org/10.1093/icb/icj015
  5. Lee S, Brown RL, Monroe W. Use of confocal laser scanning microscopy in systematics of insects with a comparison of fluorescence from different stains. Syst Entomol 2009; 34: 10-14. https://doi.org/10.1111/j.1365-3113.2008.00451.x
  6. Mair GR, Maule AG, Shaw C, Johnston CF, Halton DW. Gross anatomy of the muscle systems of Fasciola hepatica as visualized by phalloidin-fluorescence and confocal microscopy. Parasitology 1998; 117: 75-82. https://doi.org/10.1017/S0031182098002807
  7. Schawaroch V, Grimaldi D, Klaus AV. Focusing on morphology: Applications and implications of confocal laser scanning microscopy (Diptera: Campichoetidae, Camillidae, Drosophilidae). Proc Entomol Soc Wash 2005; 107: 323-335.
  8. Valdecasas AG, Abad A. Morphological confocal microscopy in arthropods and the enhancement of autofluorescence after proteinase K extraction. Microsc Microanal 2011; 17: 109-113. https://doi.org/10.1017/S1431927610094213
  9. Wilts EF, Wulfken D, Ahlrichs WH, Arbizu PM. The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology. Acta Zool 2012; 93: 14-27. https://doi.org/10.1111/j.1463-6395.2010.00476.x
  10. Worsaae K, Rouse GW.The simplicity of males: dwarf males of four species of Osedax (Siboglinidae; Annelida) investigated by confocal laser scanning microscopy. J Morphol 2010; 271: 127-142. https://doi.org/10.1002/jmor.10786
  11. Daugschies A, Bialek R, Joachim A, Mundt HC. Autofluorescence microscopy for the detection of nematode eggs and protozoa, in particular Isospora suis, in swine faeces. Parasitol Res 2001; 87: 409-412. https://doi.org/10.1007/s004360100378
  12. Morrow JJ. Exploring parasitism in antiquity through the analysis of coprolites and quids from La Cueva de los Muertos Chiquitos, Rio Zape, Durango, Mexico. (Doctoral dissertation) [dissertation]. [Lincoln]: University of Nebraska-Lincoln. 2016.
  13. Perry ES, Miller WR, Lindsay S. Looking at tardigrades in a new light: using epifluorescence to interpret structure. J Microsc 2015; 257: 117-122. https://doi.org/10.1111/jmi.12190

Cited by

  1. Paleomicrobiology of the human digestive tract: A review vol.157, 2019, https://doi.org/10.1016/j.micpath.2021.104972
  2. Exploring confocal laser scanning microscopy (CLSM) and fluorescence staining as a tool for imaging and quantifying traces of marine microbioerosion and their trace‐making microendoliths vol.284, pp.2, 2019, https://doi.org/10.1111/jmi.13046