Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow

Abstract Background Super‐resolution microscopy has enabled high‐resolution imaging of the actin cytoskeleton in megakaryocytes and platelets. These technologies have extended our knowledge of thrombopoiesis and platelet spreading using megakaryocytes and platelets cultured in vitro on matrix protei...

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Autores principales:	Yosuke Morodomi, Sachiko Kanaji, Eric Won, Tadafumi Kawamoto, Taisuke Kanaji
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2020
Materias:	bone marrow frozen sections histological techniques megakaryocytes single‐molecule imaging Diseases of the blood and blood-forming organs RC633-647.5
Acceso en línea:	https://doaj.org/article/b5f2a403af0e4fb584989a4e94159f95
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spelling	oai:doaj.org-article:b5f2a403af0e4fb584989a4e94159f952021-11-15T06:10:44ZModified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow2475-037910.1002/rth2.12276https://doaj.org/article/b5f2a403af0e4fb584989a4e94159f952020-01-01T00:00:00Zhttps://doi.org/10.1002/rth2.12276https://doaj.org/toc/2475-0379Abstract Background Super‐resolution microscopy has enabled high‐resolution imaging of the actin cytoskeleton in megakaryocytes and platelets. These technologies have extended our knowledge of thrombopoiesis and platelet spreading using megakaryocytes and platelets cultured in vitro on matrix proteins. However, for better understanding of megakaryocytopoiesis and platelet production, high‐resolution imaging of cells in an in vivo bone marrow microenvironment is required. Development of Kawamoto’s film method greatly advanced the techniques of thin cryosectioning of hard tissues such as undecalcified bones. One obstacle that remains is the spherical aberration that occurs due to the difference in the refractive index for the light path, limiting the usage of Kawamoto’s film method to lower magnification observation. Objectives To overcome the weakness of the conventional Kawamoto’s film method for higher magnification observation of undecalcified bone marrow. Methods We have modified the original method with a very simple method: flipping the film at the step of mounting the sections on the glass. Results and Conclusions This new method successfully led to the adjustment of the refractive index and enabled super‐resolution imaging of megakaryocytes in undecalcified mouse femurs. Our modified method will expand the application of Kawamoto’s film method and enable precise analysis of megakaryocytopoiesis and platelet production in the bone marrow microenvironment under pathophysiological conditions.Yosuke MorodomiSachiko KanajiEric WonTadafumi KawamotoTaisuke KanajiWileyarticlebone marrowfrozen sectionshistological techniquesmegakaryocytessingle‐molecule imagingDiseases of the blood and blood-forming organsRC633-647.5ENResearch and Practice in Thrombosis and Haemostasis, Vol 4, Iss 1, Pp 86-91 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	bone marrow frozen sections histological techniques megakaryocytes single‐molecule imaging Diseases of the blood and blood-forming organs RC633-647.5
spellingShingle	bone marrow frozen sections histological techniques megakaryocytes single‐molecule imaging Diseases of the blood and blood-forming organs RC633-647.5 Yosuke Morodomi Sachiko Kanaji Eric Won Tadafumi Kawamoto Taisuke Kanaji Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
description	Abstract Background Super‐resolution microscopy has enabled high‐resolution imaging of the actin cytoskeleton in megakaryocytes and platelets. These technologies have extended our knowledge of thrombopoiesis and platelet spreading using megakaryocytes and platelets cultured in vitro on matrix proteins. However, for better understanding of megakaryocytopoiesis and platelet production, high‐resolution imaging of cells in an in vivo bone marrow microenvironment is required. Development of Kawamoto’s film method greatly advanced the techniques of thin cryosectioning of hard tissues such as undecalcified bones. One obstacle that remains is the spherical aberration that occurs due to the difference in the refractive index for the light path, limiting the usage of Kawamoto’s film method to lower magnification observation. Objectives To overcome the weakness of the conventional Kawamoto’s film method for higher magnification observation of undecalcified bone marrow. Methods We have modified the original method with a very simple method: flipping the film at the step of mounting the sections on the glass. Results and Conclusions This new method successfully led to the adjustment of the refractive index and enabled super‐resolution imaging of megakaryocytes in undecalcified mouse femurs. Our modified method will expand the application of Kawamoto’s film method and enable precise analysis of megakaryocytopoiesis and platelet production in the bone marrow microenvironment under pathophysiological conditions.
format	article
author	Yosuke Morodomi Sachiko Kanaji Eric Won Tadafumi Kawamoto Taisuke Kanaji
author_facet	Yosuke Morodomi Sachiko Kanaji Eric Won Tadafumi Kawamoto Taisuke Kanaji
author_sort	Yosuke Morodomi
title	Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
title_short	Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
title_full	Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
title_fullStr	Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
title_full_unstemmed	Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
title_sort	modified application of kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow
publisher	Wiley
publishDate	2020
url	https://doaj.org/article/b5f2a403af0e4fb584989a4e94159f95
work_keys_str_mv	AT yosukemorodomi modifiedapplicationofkawamotosfilmmethodforsuperresolutionimagingofmegakaryocytesinundecalcifiedbonemarrow AT sachikokanaji modifiedapplicationofkawamotosfilmmethodforsuperresolutionimagingofmegakaryocytesinundecalcifiedbonemarrow AT ericwon modifiedapplicationofkawamotosfilmmethodforsuperresolutionimagingofmegakaryocytesinundecalcifiedbonemarrow AT tadafumikawamoto modifiedapplicationofkawamotosfilmmethodforsuperresolutionimagingofmegakaryocytesinundecalcifiedbonemarrow AT taisukekanaji modifiedapplicationofkawamotosfilmmethodforsuperresolutionimagingofmegakaryocytesinundecalcifiedbonemarrow
_version_	1718428577178845184

Modified application of Kawamoto’s film method for super‐resolution imaging of megakaryocytes in undecalcified bone marrow

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