Three-dimensional cellular visualization in mouse apical periodontitis using combined whole-mount staining and optical tissue clearing |
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Affiliation: | 1. Department of Cariology, Restorative Sciences, and Endodontics, University of Michigan School of Dentistry, 1011 N University Avenue, Ann Arbor, MI, 48109, USA;2. Department of Pulp Biology and Endodontics, Division of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan |
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Abstract: | Apical periodontitis is an inflammatory disease involving lesions located within the jawbone. Histological evaluations generally require decalcification and sectioning, which has limited our understanding of the three-dimensional (3D) organization and spatial distribution of different immune cell types in these lesions. A recently developed technique combining tissue clearing and whole-mount immunofluorescent labeling allows us to acquire such information from the deep tissue without sectioning. However, whole-mount immunofluorescent labeling in the jawbone requires further development. Here we provide a straightforward and efficient protocol to achieve 3D immunofluorescent imaging of murine periapical lesions. |
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Keywords: | Staining and labeling Three-dimensional imaging Periapical periodontitis Fluorescent dyes 2D" },{" #name" :" keyword" ," $" :{" id" :" kwrd0035" }," $$" :[{" #name" :" text" ," _" :" two-dimensional 3D" },{" #name" :" keyword" ," $" :{" id" :" kwrd0045" }," $$" :[{" #name" :" text" ," _" :" three-dimensional Arg1" },{" #name" :" keyword" ," $" :{" id" :" kwrd0055" }," $$" :[{" #name" :" text" ," _" :" arginase 1 BB" },{" #name" :" keyword" ," $" :{" id" :" kwrd0065" }," $$" :[{" #name" :" text" ," _" :" benzyl benzoate DDW" },{" #name" :" keyword" ," $" :{" id" :" kwrd0075" }," $$" :[{" #name" :" text" ," _" :" deionized distilled water EDTA" },{" #name" :" keyword" ," $" :{" id" :" kwrd0085" }," $$" :[{" #name" :" text" ," _" :" ethylenediaminetetraacetic acid NaCl" },{" #name" :" keyword" ," $" :{" id" :" kwrd0095" }," $$" :[{" #name" :" text" ," _" :" sodium chloride PBS" },{" #name" :" keyword" ," $" :{" id" :" kwrd0105" }," $$" :[{" #name" :" text" ," _" :" phosphate-buffered saline PEG" },{" #name" :" keyword" ," $" :{" id" :" kwrd0115" }," $$" :[{" #name" :" text" ," _" :" polyethylene glycol PEGASOS" },{" #name" :" keyword" ," $" :{" id" :" kwrd0125" }," $$" :[{" #name" :" text" ," _" :" polyethylene glycol-associated solvent system PEGMMA" },{" #name" :" keyword" ," $" :{" id" :" kwrd0135" }," $$" :[{" #name" :" text" ," _" :" poly(ethylene glycol) methyl ether methacrylate PFA" },{" #name" :" keyword" ," $" :{" id" :" kwrd0145" }," $$" :[{" #name" :" text" ," _" :" paraformaldehyde tB" },{" #name" :" keyword" ," $" :{" id" :" kwrd0155" }," $$" :[{" #name" :" text" ," _" :" tert-butanol TBST" },{" #name" :" keyword" ," $" :{" id" :" kwrd0165" }," $$" :[{" #name" :" text" ," _" :" tris-buffered saline with Triton X-100 Tris–HCl" },{" #name" :" keyword" ," $" :{" id" :" kwrd0175" }," $$" :[{" #name" :" text" ," _" :" trizma hydrochloride |
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