Loading…
Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi
Significant advances have occurred in the area of high-resolution scanning electron microscopy (SEM), especially related to methodologies that allow the observation of intracellular structures that are exposed either by successive abrasion with a gallium ion beam or by sectioning in epoxy-embedded c...
Saved in:
| Published in: | Parasitology research (1987) 2020-11, Vol.119 (11), p.3887-3891 |
|---|---|
| Main Authors: | , , |
| Format: | Article |
| Language: | English |
| Subjects: | |
| Citations: | Items that this one cites Items that cite this one |
| Online Access: | Get full text |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| cited_by | cdi_FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453 |
|---|---|
| cites | cdi_FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453 |
| container_end_page | 3891 |
| container_issue | 11 |
| container_start_page | 3887 |
| container_title | Parasitology research (1987) |
| container_volume | 119 |
| creator | de Souza, Wanderley Alcantara, Carolina L. Cunha e Silva, Narcisa L. |
| description | Significant advances have occurred in the area of high-resolution scanning electron microscopy (SEM), especially related to methodologies that allow the observation of intracellular structures that are exposed either by successive abrasion with a gallium ion beam or by sectioning in epoxy-embedded cells. Images of series of successively exposed surfaces can then be rendered into 3D models. Here, we report our observations by combining this approach with classical cytochemical methods to facilitate the 3D reconstruction of labeled structures and organelles. We used epimastigotes of
Trypanosoma cruzi
whose endocytic pathway was labeled with horseradish peroxidase, followed by fixation and detection of the peroxidase activity using the classical diaminobenzidine-osmium method followed by incubation with thiocarbohydrazide, which increases the concentration of osmium at the sites where the enzyme is located as well as the contrast of lipid-containing structures. This procedure allows not only a better visualization of membranous structures and lipid inclusions but can also easily identify the endocytic tracer (HRP) inside the cell. All structures involved in the endocytic activity could be traced and reconstructed. |
| doi_str_mv | 10.1007/s00436-020-06798-x |
| format | article |
| fullrecord | <record><control><sourceid>gale_proqu</sourceid><recordid>TN_cdi_proquest_miscellaneous_2423800984</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A639118567</galeid><sourcerecordid>A639118567</sourcerecordid><originalsourceid>FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453</originalsourceid><addsrcrecordid>eNp9kcFq3DAQhkVoaDZpXyCHIuilFycjS5blYwhpUwj0sncxK48TB1vaSDas-_TR1mkLPRQdRsx8_zAzP2OXAq4EQH2dAJTUBZRQgK4bUxxO2EYoWRaiqap3bANN_oMQ8oydp_QMIGqt1Ht2JkuthTHNhr3cDeSmGDwfexdDcmG_cLdMwT3R2KcpLhx9y6enSFS0_Ug-9cHjwCO54HN9dlNO8NDxAXc0UMvX5Bwp8d7zbVz26EMKI3IX55_9B3ba4ZDo41u8YNuvd9vb--Lhx7fvtzcPhZNNdSgaLXSrEUlKVCQN1gJ1A9iqsjLaSahkJxTVDlXb1aDMTjvlsHJalqgqecG-rG33MbzMlCab13E0DOgpzMmWqpQGoDEqo5__QZ_DHPOSR6qWRigjIVNXK_WIA9ned2GK6PJr86HyLajrc_5Gy0YIU-k6C8pVcLxritTZfexHjIsVYI8G2tVAmw20vwy0hyz69DbLvBup_SP57VgG5AqkXPKPFP8O-5-2r4yvqMQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2473814830</pqid></control><display><type>article</type><title>Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi</title><source>Springer Link</source><creator>de Souza, Wanderley ; Alcantara, Carolina L. ; Cunha e Silva, Narcisa L.</creator><creatorcontrib>de Souza, Wanderley ; Alcantara, Carolina L. ; Cunha e Silva, Narcisa L.</creatorcontrib><description>Significant advances have occurred in the area of high-resolution scanning electron microscopy (SEM), especially related to methodologies that allow the observation of intracellular structures that are exposed either by successive abrasion with a gallium ion beam or by sectioning in epoxy-embedded cells. Images of series of successively exposed surfaces can then be rendered into 3D models. Here, we report our observations by combining this approach with classical cytochemical methods to facilitate the 3D reconstruction of labeled structures and organelles. We used epimastigotes of
Trypanosoma cruzi
whose endocytic pathway was labeled with horseradish peroxidase, followed by fixation and detection of the peroxidase activity using the classical diaminobenzidine-osmium method followed by incubation with thiocarbohydrazide, which increases the concentration of osmium at the sites where the enzyme is located as well as the contrast of lipid-containing structures. This procedure allows not only a better visualization of membranous structures and lipid inclusions but can also easily identify the endocytic tracer (HRP) inside the cell. All structures involved in the endocytic activity could be traced and reconstructed.</description><identifier>ISSN: 0932-0113</identifier><identifier>EISSN: 1432-1955</identifier><identifier>DOI: 10.1007/s00436-020-06798-x</identifier><identifier>PMID: 32661889</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Abrasion ; Biomedical and Life Sciences ; Biomedicine ; Cytochemistry ; Electron microscopy ; Endocytosis ; Epimastigotes ; Epoxy resins ; Gallium ; Histocytochemistry ; Horseradish peroxidase ; Imaging, Three-Dimensional ; Immunology ; Medical Microbiology ; Microbiology ; Microscopy ; Microscopy, Electron, Scanning ; Organelles ; Organelles - ultrastructure ; Osmium ; Peroxidase ; Protozoa ; Protozoology - Short Communication ; Scanning electron microscopy ; Sectioning ; Staining and Labeling ; Trypanosoma cruzi ; Trypanosoma cruzi - metabolism ; Trypanosoma cruzi - ultrastructure</subject><ispartof>Parasitology research (1987), 2020-11, Vol.119 (11), p.3887-3891</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020</rights><rights>COPYRIGHT 2020 Springer</rights><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453</citedby><cites>FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453</cites><orcidid>0000-0002-1895-1299</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,786,790,27957,27958</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32661889$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>de Souza, Wanderley</creatorcontrib><creatorcontrib>Alcantara, Carolina L.</creatorcontrib><creatorcontrib>Cunha e Silva, Narcisa L.</creatorcontrib><title>Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi</title><title>Parasitology research (1987)</title><addtitle>Parasitol Res</addtitle><addtitle>Parasitol Res</addtitle><description>Significant advances have occurred in the area of high-resolution scanning electron microscopy (SEM), especially related to methodologies that allow the observation of intracellular structures that are exposed either by successive abrasion with a gallium ion beam or by sectioning in epoxy-embedded cells. Images of series of successively exposed surfaces can then be rendered into 3D models. Here, we report our observations by combining this approach with classical cytochemical methods to facilitate the 3D reconstruction of labeled structures and organelles. We used epimastigotes of
Trypanosoma cruzi
whose endocytic pathway was labeled with horseradish peroxidase, followed by fixation and detection of the peroxidase activity using the classical diaminobenzidine-osmium method followed by incubation with thiocarbohydrazide, which increases the concentration of osmium at the sites where the enzyme is located as well as the contrast of lipid-containing structures. This procedure allows not only a better visualization of membranous structures and lipid inclusions but can also easily identify the endocytic tracer (HRP) inside the cell. All structures involved in the endocytic activity could be traced and reconstructed.</description><subject>Abrasion</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Cytochemistry</subject><subject>Electron microscopy</subject><subject>Endocytosis</subject><subject>Epimastigotes</subject><subject>Epoxy resins</subject><subject>Gallium</subject><subject>Histocytochemistry</subject><subject>Horseradish peroxidase</subject><subject>Imaging, Three-Dimensional</subject><subject>Immunology</subject><subject>Medical Microbiology</subject><subject>Microbiology</subject><subject>Microscopy</subject><subject>Microscopy, Electron, Scanning</subject><subject>Organelles</subject><subject>Organelles - ultrastructure</subject><subject>Osmium</subject><subject>Peroxidase</subject><subject>Protozoa</subject><subject>Protozoology - Short Communication</subject><subject>Scanning electron microscopy</subject><subject>Sectioning</subject><subject>Staining and Labeling</subject><subject>Trypanosoma cruzi</subject><subject>Trypanosoma cruzi - metabolism</subject><subject>Trypanosoma cruzi - ultrastructure</subject><issn>0932-0113</issn><issn>1432-1955</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNp9kcFq3DAQhkVoaDZpXyCHIuilFycjS5blYwhpUwj0sncxK48TB1vaSDas-_TR1mkLPRQdRsx8_zAzP2OXAq4EQH2dAJTUBZRQgK4bUxxO2EYoWRaiqap3bANN_oMQ8oydp_QMIGqt1Ht2JkuthTHNhr3cDeSmGDwfexdDcmG_cLdMwT3R2KcpLhx9y6enSFS0_Ug-9cHjwCO54HN9dlNO8NDxAXc0UMvX5Bwp8d7zbVz26EMKI3IX55_9B3ba4ZDo41u8YNuvd9vb--Lhx7fvtzcPhZNNdSgaLXSrEUlKVCQN1gJ1A9iqsjLaSahkJxTVDlXb1aDMTjvlsHJalqgqecG-rG33MbzMlCab13E0DOgpzMmWqpQGoDEqo5__QZ_DHPOSR6qWRigjIVNXK_WIA9ned2GK6PJr86HyLajrc_5Gy0YIU-k6C8pVcLxritTZfexHjIsVYI8G2tVAmw20vwy0hyz69DbLvBup_SP57VgG5AqkXPKPFP8O-5-2r4yvqMQ</recordid><startdate>20201101</startdate><enddate>20201101</enddate><creator>de Souza, Wanderley</creator><creator>Alcantara, Carolina L.</creator><creator>Cunha e Silva, Narcisa L.</creator><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-1895-1299</orcidid></search><sort><creationdate>20201101</creationdate><title>Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi</title><author>de Souza, Wanderley ; Alcantara, Carolina L. ; Cunha e Silva, Narcisa L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Abrasion</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Cytochemistry</topic><topic>Electron microscopy</topic><topic>Endocytosis</topic><topic>Epimastigotes</topic><topic>Epoxy resins</topic><topic>Gallium</topic><topic>Histocytochemistry</topic><topic>Horseradish peroxidase</topic><topic>Imaging, Three-Dimensional</topic><topic>Immunology</topic><topic>Medical Microbiology</topic><topic>Microbiology</topic><topic>Microscopy</topic><topic>Microscopy, Electron, Scanning</topic><topic>Organelles</topic><topic>Organelles - ultrastructure</topic><topic>Osmium</topic><topic>Peroxidase</topic><topic>Protozoa</topic><topic>Protozoology - Short Communication</topic><topic>Scanning electron microscopy</topic><topic>Sectioning</topic><topic>Staining and Labeling</topic><topic>Trypanosoma cruzi</topic><topic>Trypanosoma cruzi - metabolism</topic><topic>Trypanosoma cruzi - ultrastructure</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>de Souza, Wanderley</creatorcontrib><creatorcontrib>Alcantara, Carolina L.</creatorcontrib><creatorcontrib>Cunha e Silva, Narcisa L.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Parasitology research (1987)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>de Souza, Wanderley</au><au>Alcantara, Carolina L.</au><au>Cunha e Silva, Narcisa L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi</atitle><jtitle>Parasitology research (1987)</jtitle><stitle>Parasitol Res</stitle><addtitle>Parasitol Res</addtitle><date>2020-11-01</date><risdate>2020</risdate><volume>119</volume><issue>11</issue><spage>3887</spage><epage>3891</epage><pages>3887-3891</pages><issn>0932-0113</issn><eissn>1432-1955</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>Significant advances have occurred in the area of high-resolution scanning electron microscopy (SEM), especially related to methodologies that allow the observation of intracellular structures that are exposed either by successive abrasion with a gallium ion beam or by sectioning in epoxy-embedded cells. Images of series of successively exposed surfaces can then be rendered into 3D models. Here, we report our observations by combining this approach with classical cytochemical methods to facilitate the 3D reconstruction of labeled structures and organelles. We used epimastigotes of
Trypanosoma cruzi
whose endocytic pathway was labeled with horseradish peroxidase, followed by fixation and detection of the peroxidase activity using the classical diaminobenzidine-osmium method followed by incubation with thiocarbohydrazide, which increases the concentration of osmium at the sites where the enzyme is located as well as the contrast of lipid-containing structures. This procedure allows not only a better visualization of membranous structures and lipid inclusions but can also easily identify the endocytic tracer (HRP) inside the cell. All structures involved in the endocytic activity could be traced and reconstructed.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>32661889</pmid><doi>10.1007/s00436-020-06798-x</doi><tpages>5</tpages><orcidid>https://orcid.org/0000-0002-1895-1299</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0932-0113 |
| ispartof | Parasitology research (1987), 2020-11, Vol.119 (11), p.3887-3891 |
| issn | 0932-0113 1432-1955 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2423800984 |
| source | Springer Link |
| subjects | Abrasion Biomedical and Life Sciences Biomedicine Cytochemistry Electron microscopy Endocytosis Epimastigotes Epoxy resins Gallium Histocytochemistry Horseradish peroxidase Imaging, Three-Dimensional Immunology Medical Microbiology Microbiology Microscopy Microscopy, Electron, Scanning Organelles Organelles - ultrastructure Osmium Peroxidase Protozoa Protozoology - Short Communication Scanning electron microscopy Sectioning Staining and Labeling Trypanosoma cruzi Trypanosoma cruzi - metabolism Trypanosoma cruzi - ultrastructure |
| title | Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi |
| url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-09-23T04%3A27%3A49IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_proqu&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Electron%20microscopy%20cytochemistry%20and%20three-dimensional%20reconstruction%20of%20labeled%20structures%20in%20Trypanosoma%20cruzi&rft.jtitle=Parasitology%20research%20(1987)&rft.au=de%20Souza,%20Wanderley&rft.date=2020-11-01&rft.volume=119&rft.issue=11&rft.spage=3887&rft.epage=3891&rft.pages=3887-3891&rft.issn=0932-0113&rft.eissn=1432-1955&rft_id=info:doi/10.1007/s00436-020-06798-x&rft_dat=%3Cgale_proqu%3EA639118567%3C/gale_proqu%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c395x-9616d6aae33a4e38a71a690ad42586c3053f14e7ca4df7048b6c4ca5c632a453%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2473814830&rft_id=info:pmid/32661889&rft_galeid=A639118567&rfr_iscdi=true |