{"id":467,"date":"2022-04-12T17:00:00","date_gmt":"2022-04-12T08:00:00","guid":{"rendered":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/?page_id=467"},"modified":"2023-12-19T08:51:10","modified_gmt":"2023-12-18T23:51:10","slug":"paper","status":"publish","type":"page","link":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/paper.html","title":{"rendered":"\u5b66\u8853\u8ad6\u6587\u3067\u306e\u5229\u7528\u6a5f\u5668\u3084\u8b1d\u8f9e\u306e\u66f8\u304d\u65b9"},"content":{"rendered":"\n

\u5b66\u8853\u8ad6\u6587\u306e”Material and Methods”\u3067\u306e\u8a18\u8f09\u65b9\u6cd5.<\/h2>\n\n\n\n

\u5b66\u8853\u8ad6\u6587\u3067\u306e\u3001\u5229\u7528\u6a5f\u5668\u306e\u8a18\u8ff0.<\/h3>\n\n\n\n

\u8ad6\u6587\u306e”Material and Methods”\u306a\u3069\u3067\u5229\u7528\u6a5f\u5668\u3092\u8a18\u3059\u5834\u5408\u3001\u4ee5\u4e0b\u306e\u3088\u3046\u306b\u6a5f\u5668\u306e\u8a73\u7d30\u3092\u8a18\u3059\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002(\u3053\u306e\u4f8b\u306f\u30012023\u5e7410\u6708\u73fe\u5728\u3067\u306eStation-3\u306e\u4ed5\u69d8\u3067\u3059)\u3002\u7279\u306b\u3001\u5bfe\u7269\u30ec\u30f3\u30ba\u306e\u8a18\u8f09<\/strong><\/span>\u306f\u3001\u9855\u5fae\u93e1\u89b3\u5bdf\u306e\u5834\u5408\u306f\u5fc5\u9808\u3067\u3059\u3002<\/p>\n\n\n\n

An inverted microscope (Ti-E, Nikon) equipped with a Plan Apo VC \u00d760 objective lens (NA 1.40, Nikon) and micro scanning stage (BI XY stage, Chuo Precision Industrial Co. Ltd.) was used to observe fluorescence images in living cells maintained at 37\u00b0C with a continuous supply of 95% air and 5% carbon dioxide by using a stage-top incubator (STXG-WSKMX-SET, Tokai Hit). Images were taken by scientific Complementary Metal Oxcide Semiconductor (sCMOS) camera (ZL41 Wave, Oxford Instruments).

\u203b2023\u5e74\uff19\u6708\u306b\u3001\u30ab\u30e1\u30e9\u304c\u4ea4\u63db\u3068\u306a\u308a\u307e\u3057\u305f\u3002\u305d\u308c\u4ee5\u524d\u306e\u5229\u7528\u306e\u5834\u5408\u306f\u3001\u30ab\u30e1\u30e9\u306e\u6a5f\u7a2e\uff08\u672b\u5c3e\u306e\u30ab\u30c3\u30b3\u90e8\u5206\uff09\u3092\u3001\uff08Zyla 5.5, Oxford Instruments\uff09\u3068\u3001\u3054\u8a18\u5165\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

\u307e\u305fStation-1, Station-4\uff08Nikon:A1\uff09\u3067\u306e\u5171\u7126\u70b9\u89b3\u5bdf\u306e\u5834\u5408\u3082\u3001”\u30e1\u30a4\u30f3\u306f\u9855\u5fae\u93e1\u3067\u3001\u305d\u3053\u306b\u5171\u7126\u70b9\u30b7\u30b9\u30c6\u30e0\u304c\uff08\u5468\u8fba\u6a5f\u5668\u3068\u3057\u3066\uff09\u5b9f\u88c5\u3055\u308c\u3066\u3044\u308b<\/strong><\/span>“\u3053\u3068\u306b\u306a\u308a\u307e\u3059\uff08\u9855\u5fae\u93e1\u3084\u5bfe\u7269\u30ec\u30f3\u30ba\u304c\u306a\u3044\u3068\u3001\u753b\u50cf\u304c\u64ae\u308c\u307e\u305b\u3093\uff09\u3002
\u524d\u8ff0\u3068\u540c\u69d8\u306b\u3001\u4ee5\u4e0b\u306e\uff12\u4f8b\u306e\u3088\u3046\u306b\u9855\u5fae\u93e1\u3084\u5bfe\u7269\u30ec\u30f3\u30ba\u306e\u8a18\u8f09\u304c\u5fc5\u8981\u3067\u3059\u304c\u3001\u4e00\u65b9\u3067\u5171\u7126\u70b9\u9855\u5fae\u93e1\u306e\u5834\u5408\u306f\u3001\u5149\u5b66\u30d5\u30a3\u30eb\u30bf\u30fc\u306e\u8a18\u8f09\u306f\u6982\u306d\u4e0d\u8981\u3067\u3059\u3002<\/p>\n\n\n\n

Confocal images were taken by a confocal laser scanning microscopy system (Nikon A1 and Ti-E) equipped with a Plan Apo VC x60 objective lens (NA 1.40, Nikon). In order to maintain condition, a stage-top incubator (STXG-WSKMX-SET, Tokai Hit) was set onto microscope stage for live cell imaging.

A Nikon Ti-E inverted microscope equipped with a Nikon A1Rsi spectral imaging confocal scanning system was used for confocal microscopy experiments.<\/p>\n\n\n\n

\u8ad6\u6587\u3067\u306e\u6a5f\u5668\u8a73\u7d30\u306e\u66f8\u304d\u65b9\u306f\u3001\u30d5\u30a3\u30eb\u30bf\u30fc\u3084\u5bfe\u7269\u30ec\u30f3\u30ba\u306a\u3069\u3082\u542b\u3081\u305f\u89b3\u5bdf\u72b6\u6cc1\u306e\u5165\u5ff5\u306a\u78ba\u8a8d\u3082\u5fc5\u8981\u3068\u306a\u308a\u3001\u66f4\u306b\u306f\u5b66\u8853\u8a8c\u306b\u3088\u3063\u3066\u3001\u8a18\u8f09\u5185\u5bb9\u304c\u7570\u306a\u3063\u3066\u3044\u307e\u3059\u3002\u4e00\u4f8b\u3068\u3057\u3066\u3001\u300c\u30e1\u30fc\u30ab\u30fc\u306e\u6240\u5728\u90fd\u5e02\u300d\u306e\u8a18\u8f09\u3082\u5fc5\u8981\u306a\u5b66\u8853\u8a8c\u3082\u3042\u308b\u306a\u3069\u3001\u9069\u5207\u306a\u8a18\u8f09\u65b9\u6cd5\u304c\u76f4\u3061\u306b\u306f\u308f\u304b\u3089\u306a\u3044\u304b\u3082\u3057\u308c\u307e\u305b\u3093\u3002
\u3054\u4e0d\u660e\u306e\u70b9\u306a\u3069\u304c\u3054\u3056\u3044\u307e\u3059\u306a\u3089\u3001\u3044\u3064\u3067\u3082\u304a\u554f\u3044\u5408\u308f\u305b\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

Huygens\u3092\u4f7f\u3063\u305f\u969b\u306e\u8a18\u8ff0.<\/h3>\n\n\n\n

Huygens\u306b\u3088\u308b\u30c7\u30b3\u30f3\u30dc\u30ea\u30e5\u30fc\u30b7\u30e7\u30f3\u51e6\u7406\u3092\u884c\u3063\u305f\u969b\u306e\u6587\u4f8b\u3092\u8f09\u305b\u3066\u304a\u304d\u307e\u3059\u3002\u8ad6\u6587\u3092\u66f8\u304b\u308c\u308b\u969b\u306b\u306f\u3001\u3069\u3046\u305e\u53c2\u8003\u3068\u3057\u3066\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

\n

For deconvolution microscopic image, serial optical section data (20-30 focal planes at 0.5\u03bcm intervals) were collected and computationally processed by a 3D deconvolution method (Huygens essential deconvolution software) to remove out-of-field fluorescence.<\/p>\n\n\n\n

All images from fixed cells were captured using AquaCosmos (Hamamatsu Photonics) and processed through the Huygens Essential Deconvolution software (SVI, Hilversum, Netherlands) using the Classic Maximum Likelihood Estimation method.<\/p>\n<\/div><\/div>\n\n\n\n

\u5b66\u8853\u8ad6\u6587\u3067\u306e\u8b1d\u8f9e\u306e\u66f8\u304d\u65b9<\/h2>\n\n\n\n

\u30a4\u30e1\u30fc\u30b8\u30f3\u30b0\u30bb\u30f3\u30bf\u30fc\u306b\u5bfe\u3057\u3066\u306e\u8b1d\u8f9e<\/h3>\n\n\n\n

\u4ee5\u4e0b\u306b\u306f\u3001\u5b66\u8853\u8ad6\u6587\u3067\u306e\u8b1d\u8f9e\u306e\u6587\u4f8b\u3092\u8f09\u305b\u3066\u304a\u304d\u307e\u3059\u304c\u3001\u3053\u306e\u3088\u3046\u306a\u4e00\u822c\u7684\u306a\u66f8\u304d\u65b9\u3067\u5341\u5206\u3067\u3059\u3002
\u8ad6\u6587\u3092\u66f8\u304b\u308c\u308b\u969b\u306b\u306f\u3001\u3069\u3046\u305e\u53c2\u8003\u3068\u3057\u3066\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

\n
\n

We are grateful to the Nikon Imaging Center at Hokkaido University for being very helpful with confocal microscopy, image acquisition, and analysis.<\/p>\n<\/div><\/div>\n\n\n\n

We would like to thank the Nikon Imaging Center at Hokkaido University for technical support.<\/p>\n\n\n\n

The authors would like to thank the Nikon imaging Center at Hokkaido University for imaging equipment and software.<\/p>\n\n\n\n

Confocal images were acquired in the Nikon Imaging Center at Hokkaido University.<\/p>\n\n\n\n

Microscopy analysis of samples was performed in the Nikon Imaging Center at Hokkaido University, using a Nikon Ti-E inverted microscope.<\/p>\n<\/div><\/div>\n\n\n\n

\u5148\u7aef\u30d0\u30a4\u30aa\u30a4\u30e1\u30fc\u30b8\u30f3\u30b0\u652f\u63f4\uff08ABiS\uff09\u306e\u652f\u63f4\u306b\u5bfe\u3057\u3066\u306e\u8b1d\u8f9e<\/h3>\n\n\n\n

ABiS\u306e\u652f\u63f4\u3092\u53d7\u3051\u3066\u3044\u308b\u65b9\u306f\u3001ABiS\u3078\u306e\u8b1d\u8f9e\u3082\u3001\u4ee5\u4e0b\u306e\u3088\u3046\u306a\u5f62\u3067\u3054\u8a18\u5165\u304f\u3060\u3055\u3044\u3002\u7b2c\uff11\u671f\u3068\u7b2c\uff12\u671f\u3067\u3001\u79d1\u7814\u8cbb\u756a\u53f7\u3082\u542b\u3081\u3066\u3001\u5c11\u3057\u8a18\u8f09\u304c\u7570\u306a\u3063\u3066\u304a\u308a\u307e\u3059\u306e\u3067\u3001\u3054\u6ce8\u610f\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

\u307e\u305f\u5b66\u8853\u8ad6\u6587\u304c\u767a\u8868\u3068\u306a\u308a\u307e\u3057\u305f\u3089\u3001\u4f75\u305b\u3066ABiS\u4e8b\u52d9\u5c40\u3078\u306e\u3054\u5831\u544a<\/a>\u3082\u304a\u9858\u3044\u3044\u305f\u3057\u307e\u3059\u3002\u3053\u3061\u3089\u306b\u3064\u3044\u3066\u306f\u3001\u30d5\u30a9\u30fc\u30e0\u304b\u3089\u7c21\u5358\u306b\u5831\u544a\u3067\u304d\u307e\u3059\u3046\u3048\u3001ABiS\u306eWeb\u30b5\u30a4\u30c8\u306a\u3069\u3067\u3082\u5e83\u304f\u30a2\u30d4\u30fc\u30eb\u3057\u3066\u304f\u3060\u3055\u308a\u307e\u3059\u3002<\/p>\n\n\n\n

\n
\n

2022\u5e74\u5ea6-2027\u5e74\u5ea6\u307e\u3067\u306e\u652f\u63f4\u5bfe\u8c61\u8005<\/span><\/span><\/strong><\/p>\n<\/div><\/div>\n\n\n\n

This work was supported by JSPS KAKENHI Grant Number JP22H04926<\/span>.<\/p>\n\n\n\n

This work was supported by JSPS KAKENHI Grant Number JP22H04926<\/span>, Grant-in-Aid for Transformative Research Areas \u2015 Platforms for Advanced Technologies and Research Resources \u201cAdvanced Bioimaging Support\u201d.<\/p>\n<\/div><\/div>\n\n\n\n

\n
\n

2016\u5e74\u5ea6-2021\u5e74\u5ea6\u307e\u3067\u306e\u652f\u63f4\u5bfe\u8c61\u8005<\/span><\/span><\/strong><\/p>\n<\/div><\/div>\n\n\n\n

This work was supported by JSPS KAKENHI Grant Number JP16H06280<\/span>.<\/p>\n\n\n\n

This work was supported by JSPS KAKENHI Grant Number JP16H06280<\/span>,\u3000Grant-in-Aid for Scientific Research on Innovative Areas \u2015 Platforms for Advanced Technologies and Research Resources \u201cAdvanced Bioimaging Support\u201d.<\/p>\n<\/div><\/div>\n\n\n\n\n","protected":false},"excerpt":{"rendered":"

\u5f53\u30bb\u30f3\u30bf\u30fc\u306e\u9855\u5fae\u93e1\u3092\u5229\u7528\u306b\u3042\u305f\u308a\u3001\u8ad6\u6587\u3067\u306e\u88c5\u7f6e\u306e\u8a18\u8f09\u3084\u8b1d\u8f9e\u306e\u4e00\u4f8b\u3092\u8a18\u3057\u307e\u3059\u3002<\/p>\n","protected":false},"author":2,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"swell_btn_cv_data":""},"_links":{"self":[{"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/pages\/467"}],"collection":[{"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/comments?post=467"}],"version-history":[{"count":13,"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/pages\/467\/revisions"}],"predecessor-version":[{"id":1755,"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/pages\/467\/revisions\/1755"}],"wp:attachment":[{"href":"https:\/\/c-mng.cwh.hokudai.ac.jp\/nic.es\/Root\/wp-json\/wp\/v2\/media?parent=467"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}