Application of Autofluorescence for Confocal Microscopy to Aid in Archaeoparasitological Analyses
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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.
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.
참고문헌 (13)
- Böhm A, "Confocal imaging of the exo-and endoskeleton of Protura after nondestructive DNA extraction", Soil Organisms, 83, pp.335~345 (2011)
- Chetverikoc PE, "Confocal laser scanning microscopy technique for the study of internal genitalia and external morphology of eriophyoid mites(Acari : Eriophyoidea)", Zootaxa, 3453, pp.56~68 (2012)
- Grzywacz A, "Confocal laser scanning microscopy as a valuable tool in Diptera larval morphology studies", Parasitol Res, 113, pp.4297~4302 (2014)
- Klaus AV, "Novel methodology utilizing confocal laser scanning microscopy for systematic analysis in arthropods(Insecta)", Integr Comp Biol, 46, pp.207~214 (2006)
- Lee S, "Use of confocal laser scanning microscopy in systematics of insects with a comparison of fluorescence from different stains", Syst Entomol, 34, pp.10~14 (2009)
- Mair GR, "Gross anatomy of the muscle systems of Fasciola hepatica as visualized by phalloidin-fluorescence and confocal microscopy", Parasitology, 117, pp.75~82 (1998)
- Schawaroch V, "Focusing on morphology : Applications and implications of confocal laser scanning microscopy(Diptera : Campichoetidae, Camillidae, Drosophilidae)", Proc Entomol Soc Wash, 107, pp.323~335 (2005)
- Valdecasas AG, "Morphological confocal microscopy in arthropods and the enhancement of autofluorescence after proteinase K extraction", Microsc Microanal, 17, pp.109~113 (2011)
- Wilts EF, "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, 93, pp.14~27 (2012)
- Worsaae K, "The simplicity of males : dwarf males of four species of Osedax(Siboglinidae; Annelida)investigated by confocal laser scanning microscopy", J Morphol, 271, pp.127~142 (2010)
- Daugschies A, "Autofluorescence microscopy for the detection of nematode eggs and protozoa, in particular Isospora suis, in swine faeces", Parasitol Res, 87, pp.409~412 (2001)
- 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, pp.2016
- Perry ES, "Looking at tardigrades in a new light : using epifluorescence to interpret structure", J Microsc, 257, pp.117~122 (2015)
