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1章 光学顕微鏡と電子顕微鏡
ƩƲžŷõ ƜȘÝ ¥õȏĞȀ*ȋóȏĞȀ !*# * ƩƲžŷõ*¥õȏĞȀ ƩƲ/řƳCƎ?)!/ŕǭCƎ?0ÎŚ)?ȗ ƩƲžŷõ0¥õȏĞȀ/ƅń**:-ñ6>ʼn,? ŀīLJ/ÿ¦*-ǮĄ#/ǰ©0:&(ǸǍ ,ę»CŐ#(?ȗ ȏĞȀ/ŞÐ ȏĞȀ01590ďA-K|^/ƌȀí)?Hans Jansen and Hooke /ȏĞȀ Ȕhttp:// www.pyonta.city.hiroshima.jp/blog/ pages/number/268.htmlȕ Lacharias Jansen *ǐó-<&(ƅń@# 1655ďGO}X/ Robert Hooke 0ýŷY*ĵƌY) ŕħ@? njēȏĞȀCNj 1665ď-0!/ȏĞȀC&(66,žŷCǑü#Ǖǿ ȏĞȀßǜȔMicrographiaȕCƅLj Robert Hooke0/Ǖǿ/)ƩƲì'?Ā,Űļ/ ǴăC”ƩƲȔCellȕ”*Ô# !ƩƲǙ Cell DoctorineȔ1838ȕ K|^/Antonie van Leeuwenhoek 0ÅYCſ# Åēȏ ĞȀCNj 1673ď-ǠDžż/ôâCƅǎ=-Ʃƽ;\H}wW (1677) ƣó:ƅǎ# 1931ďeGa/ Max Knoll *Ernst A.F. Ruska ǪǯäȋóȏĞȀC Ȃƅ Leeuwenhoek /ȏĞȀ Ȕhttp://teslabar.blogspot.jp/2011/07/blog- post_03.htmlȕ ƩƲ/Ǒü¥õȏĞȀ*ȋóȏĞȀ !! ƩƲ/Ǒü ƩƲ/Ǒü¥õȏĞȀ*ȋóȏĞȀ ǕļŤ 1024 1021 1018 1015 1012 109 106 103 102 101 Y Z E P T G M K H D ȏĞȀ/ýǝơÞ Yotta Zetta Exa Peta Tera Giga Mega Kilo Hecto Deca 10-1 10-2 10-3 10-6 10-9 10-12 10-15 10-18 10-21 10-24 d c m µ n p f a z y deci centi milli micro nano pico femto atto zepto yocto ƩƲ/Ǒü¥õȏĞȀ*ȋóȏĞȀ )0+6)ǎ?/ȡ ´ǓƳResolution : 2 '/ŷ¶-ǎ?ŊĀ/ȃȆ 0.61 " ´ǓƳ= n sin# Fraunhofer /ÝĮē W = 0.61/ȂÌÄĚ }Q/ŊĀŮC Rayleight ´ǓƳġſ# http://www.tuat.ac.jp/~katsuaki /el/ EL2008/slide_hikari04.pdf " = ſ?¥/ťȁ n sin # = ȂÌļ n = ǞĽ*ýŷȔȉ¥ȕY /ȃ-?òǟ/ĂĮź # = ǖĽ/ Ů=ýŷY -<&(ȉ9=@?¥/Ȏǒ /Ä´/ǒĒ ưƌ/´ǓƳ0Ȗƥ 0.1 mmȔǏƩƲ/ð (2~3 µm) -ô?ȕ ¥õȏĞȀ)0Ȗƥ 0.2 µmȔ" = 400 nmȖn sin # = 1.4ȕ Ğ´čŨȏĞȀ*Sm{]Ǔŏ)0.025 µm 6)ÎƳ ȋóȏĞȀ)0Ȗ" = 0.004 nm )?/)ȖŽǛ0Ȗƥ 0.002 nm *ǔƞ@?ùâ?ȏĞȀ0, 0.1 nm ȄƁ ƩƲ/Ǒü¥õȏĞȀ*ȋóȏĞȀ !! ¥õȏĞȀ ȔÎǏ¥ťȁ 400~700 nm ;ƨïƮȕ !! ńǏǹȏĞȀ !! ƊĉȏĞȀ !! Ğ´čŨȏĞȀ !! ňǏǹȏĞȀ !! ¥ȏĞȀ !! ǃ¥ȏĞȀ ¨űŮVȏĞȀ !! ȋóȏĞȀ ȔȋóƮťȁ 0.004 nmȢ0.002 nmȕ !! 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Ğ´čŨȏĞȀ Ğ´čŨȏĞȀ-0ScU *ýŷY !@"@Ğ´čŨp}YwȔDICp}Ywȕ? !/2'/p}YwCij8<-2'/¥Ŏ u||GV* Ef|GVǵƯ@(? ¥ū=/¥0u||GVCǪǯ( ŁÕ-Ĵ¿ ?¥Ȕ¥ȕ-îķ@? /¥0ScU /DICp}Yw-<&( ŁÕ-Ĵ¿?2ŋ/¥-´=@45Ďdž-U p~-Ė#? 2ŋ/¥0B-Ǯǥ@(?# 9Up~/ÆîÂ(?ǴCǫ?*0Up~ªCǫǯ?¥/ǬĒ/Dz-<>¥ǥ ĉȔ¥õƇǤȊ/ĉȕCž? /2ŋ/¥Ȕabȕȃ-¥ǥĉ?*čŨ* ŻǝǢ>ńň/Sd|Xdž? /<-Ğ´čŨȏĞȀ0Up~/Æ/Àǵ -<?¥ǥĉCSd|Xd-îķ?*)Ǫń ,Up~/ǑüCÎƳ-(? ëĕ ũĔ ǃ¥ȏĞȀ ǃ¥&(,-ȡ 収縮胞 核 微小管 食胞(DV-1) トリコシスト ¥Ih~OCËÊ?*-<>½Ǣ@#ŷǟç/ŹĦ -:+?Ņ-Ǣ?ƅ¥/*C¥~vh`ZX* ǃ¥0/ Ɩ)? ½ǢŹĦ ½Ǣ¥ ǃ¥ e- çđŹĦ ŷǟ/´ó?0Çó¥Ih~OC×É?* ŷǟ/´ó?0Çó ¥Ih~OC×É?* !@6)-öø#ŹĦȔçđŹĦȕCć@öø,ŹĦ Ȕ½ǢŹĦȕ-,? /´,Ih~O0ĺ³@?*)öø-Ĩ?*) ?ȗ/Ņĺ³@?/ǃ¥)?ȗ マイオネーム ǃ¥ȏĞȀ ǃ¥ȏĞȀ-0Ÿø/¥Cǫǯ ?ț'/nF~]? ¥ū=0ƨïƮ;ÎǏ¥C§(Ö 9#¥Ĭþ@?/)@C¸Ȅ ?ĠǍ? șƂƈ/nF~]0½ǢIh~O C?ťȁC¸Ȅ?nF~]) ?Ȕ½ǢnF~]ȕ ȚƂƈ/nF~]0Ÿø/¥/7C Èþ ?nF~])? Ȕ^GPG`Pv|ȕ /nF~]-<>½Ǣ¥0§(ǖ Ľ-Èþ@ĵƌY)Ǒü@ ?*0, țƂƈ/nF~]0½Ǣ¥-<>ĺ ³@#Ÿøťȁ/ǃ¥/7CǑü ?#9/nF~])? Ȕ×ÉnF~]ȕ '6>½Ǣ¥)B@#ťȁCL`d ?nF~])? 収縮胞 核 微小管 食胞(DV-1) トリコシスト マイオネーム http://www.nikon-instruments.jp/jpn/PRODUCTS/OPTION/INDEX1.ASPX ǃ¥ȏĞȀ ǃ¥ƼƧ&(ȡ 収縮胞 ƺƇ-·ſ@(?Đ'/ǃ¥ƼƧ /½Ǣťȁ<2ǃ¥ťȁCÑ/ß-Ɠ ƼƧ´ó=ĺ³@?ǃ¥Ih~O0 ĠŲƇ-=@#½ǢIh~O<> ,? /#9½Ǣťȁ<>:ǃ¥ťȁ0ȁ ,?*ťȁ/ĉž? DAPI: DNACő9?ƼƧ GFP: green fluorescence protein KP|R=į³@#ǃ¥ǂƆǟ CFP, YFP: GFP CĹƻ#ǃ¥ǂƆǟ FITC: fluorescein isothiocyanate ƺƇ-ƻB@?ƭƼǃ¥ŷǟ Rhodamin: Phalloidin *ƪÒ (EP_ CőƼ?Ņ-ƻB@? Alexa: Alexa Fluor ÚÙÔ ǃ¥ĕĒıƬģ*:-¢@? RFP: red fluorescence protein UTƀō/ǂƆǟ Cy3, 5: cyanine ƼƧ 核 微小管 食胞(DV-1) トリコシスト マイオネーム ǃ¥ȏĞȀȠ¨űŮVȏĞȀ ¥ū-V¥Cǃ¥ȏĞȀ V¥&(,-ȡ Light Amplification by Stimulated Emission of Radiation = ĺ³ǘÿ#¥/ëČ = ĈƇ,¥ = Å /ťȁÓƊCı'¥İĻ,ȔIJ¥ģȒȕ http://home.s00.itscom.net/large/ELEC/laser/index.html http://home.s00.itscom.net/large/ELEC/laser/index.html ǫċ/¥0ǮdžŁÕÓ)0,/ )YC'&(:§( Ů-ȉ9 ?*), V¥0ǮdžŁÕƊØťļ Ķ&(>Y-<>Ȍċ-Ā, Ů-ȉ9?*)? /#9Ih~OûĒȌċ-Ȓ http://bioimaging.jp/learn/023/ ¨űŮVȏĞȀ )¨űŮ&(ȡ ½Ǣ¥)? V¥/ űŮ Ǒü?Ů/ űŮ =B?C@ @= ƹ ?Ů$C Ǒü?* )? AB< ( =B?C@+63257* 1;B>@3" $,%-98 ;B>@3 +63"4' =B?C@:&.0 /8 ) muGd )Ǒü? ĤèȉƗĠ Ǎ*,? ¨űŮVȏĞȀ ŮC+;&(¡-?ȡ ǡŒscaninng *&(/ß/<-Ãȓ-#+>ȍC'? V)0űŮ/ƫ>Ł)ŽǛƇ-0 űŮ/ƫ>Ł)ŽǛƇ-0 nm r~)/Ǒü ÎƳ)? /0 XY Ďȍ-Ȅ= Z ǧŁÕ:ÓƣĒ)Ǒü)? (ĝ=@#ȍCŢĆȉƗ?*)ƚ/¬ŕƢ? http://bioimaging.jp/learn/023/ ǃ¥ȏĞȀ ,D/ę-ƚ'.Dȡ șȕǃ¥őƼºCſ(Ÿø/ƩƲ ªĀÜ÷Cő9´? DAPI œőƼ vdSe}E/ȋôģőƼ ƩƲªǷģÂǴ/Óø ȚȕŸø/]kPǟ/ƩƲªāâ CƑǗ? ȃĵǃ¥ĭŤ GFP*DŽÒ ?ėǟǦķ țȕŸø/dzó/őƼ)/´Ċ CƑǗ? Fluorescence in situ Hybridization ȜȕǂƆǟ/Ɣ¿ǬĒ/Ǔŏ Fluorescent Recovery after Photobreach ȋóȏĞȀ &(,-ȡ ȏĞȀ*<>0ųƙ/ð,Ō ȏĞȀ*<>0 /«Ɲä/:/0ȋóǽC¤ƛ-:& /«Ɲä/:/0 #ȋƒSG~/ȉ6> #ȋƒSG~ /Ɲ/ªǴ0ȒƋƘ-,&(? ȋóǽ=0~100 kV /ȋáC(ȋ ȋóǽ=0 óƮCȐ1 ȋóƮ/ťȁ0ƥ ~0.004 nm )>ȋ ȋóƮ/ťȁ0 óƮC¥ū*? óƮC 6#ȋƒSG~Y/ę»C? ǪǯäȋóȏĞȀ = Transmission Electron Microscope (TEM) ǖĽ-ȋóƮC(ǖĽCǫǯ(ō# ȋóCȋƒSG~)ƪ¡ Ǒü?ȏ ĞȀ/*)? Ƙş)0ȋó0Ƹƀ-¿,/) ªǴ0ȒƋƘ-#@(? ȔȅŔƟ/<,:/ȕ ¥õȏĞȀ/éÒ-:¥CǪǯ ü?ÇŽƇ-0ƻ(? (Ǒ ǫǯ#ȋó0đǴ-#ǃ¥Ŏ)Ǒ ü? ǖĽ0Ĝ&(ȋóǫ?ƕǁħ? 6#ǖĽ-0ǸǺąC&#Âõàø; ȋóǪǯ³ō,Ŗ,ǸǺąC:%# ȋóőƼCłĠǍ? ǪǯäȋóȏĞȀ TEM ǖĽħ/Ŧ@ șȕÂõàø: ĕàøº/Q~]~E~cle ǸǺąCÖ8KXvHwǷOsO4 Cſ ȚȕƵŠȠŠŬŧ/ǖĽCƴŬģ/Řƴ-Áå ?ůI]i~;pmKNUG e-Ưķ țȕŘƴÁåȠIuNWƤŘƴȖQuetol 812 Řƴ/ƐÂ80 over night ȜȕǣǁµŶħȠM|XfGnor^Gzyef GnCZ`d#vPdwCſ(Æ ƥ90 nm ǣǁµŶCĝ? ǾNj/x`W{-ǣǁµŶCǨ ? ȝȕȋóőƼȠǶǷH|g~ŠŬŧ*ƏǷǻŬŧ 2 ǸőƼCdžȗőƼ0úŪ) ȞȕǑüȠx`W{CǪǯäȋóȏĞȀ-Z`d 75 kV -(ǑüȖƋĸĘCdž ǡŒäȋóȏĞȀ = Scanning Electron Microscope (SEM) 6#:;ȏĞȀ*<>0ĀĴ>, ųƙ/ð,Ō /«Ɲä/:/0ȋóǽC¤ƛ-:& #ȋƒSG~/ȉ6> ȋóǽ=0~10 kV /ȋáC(ȋó ƮCȐ1 TEM */Dz0ȋóCǪǯ ?/)0 ,Èþ(ō#ȚśȋóCǑü? ƚƇ,¡Ǒü@? ǡŒäȋóȏĞȀ SEM ǖĽħ/Ŧ@ șȕÂõàø: ĕàøº/Q~]~E~cle ǸǺąCÖ8KXvHwǷOsO4 Cſ ȚȕƵŠȠI]i~Cſ? țȕŵȠCO2 ƷƁŮŵ;t-o_~E~S~ -Ưķ(¯ƪŵ? ȜȕǖĽÏ3/ĵƍȠÿȋģ/Ltȍbp ;ǼsXdC&(E~v/ǖĽÏ -ĵƍ? ȝȕÿȋ°ŽȠƋƘǀƍ;GKXk`]}Q<>Ǻ;ƆǺCǖĽLjȍ-SbQ? @-<>Țśȋó/ĺ³¾źC? ȞȕǑüȠx`W{CǪǯäȋóȏĞȀ-Z`d 5 kV ƕĒ-(ǑüȖƋĸĘCdž ȟȕǡŒȋóȏĞȀ0¨űŮ/Ņ-Ǚń#< -ȋóƮCÃȓ-Ŵþ?*)ŷ /²±/ĤèCĝ(? http://hr-inoue.net/zscience/topics/sem/sem.html ǪǯäȋóȏĞȀ,/-ƚƇ n}Yp}LŤ ĢǬ¯ƪŤ-<>ŰÂõàø/ŗŋǑü CÎƳ*?īLJ ¯ƪ»Ŀ-<>Ĝō/µŶŤ)0ĝ=@ ,&#ƶª/ś£ŕǭǑüÎƳ* ,>ƩƲžŷõ-oPX~C: #=# ¯ƪ#ƩƲC Lv[}/ŝ) »Ŀ?* ģǟ/ƃ,?Ǵ ´)lj? ƩƲCǺnJG~ /)æȑ? /ƩƲCȀ/< -ŭ=,Ljȍ Cı'Ǿo`P /-Í? Ǿ09ŧq} HwȔ269!ȕ )Ȗ®;(? ƋƘ)90! ƕĒ-Ň9šCŃƾ ȔI`_Qȕ ??0»Ŀ Ɖě-ƆǺCǀƍ p}LC ħ? http://www.tech.nagoya-u.ac.jp/event/h23/Vol07/hon_secur/OSEI-1_s.pdf n}Yp}LŤ ǡŒä/ǞĽħ)Ǚń #<-ƆǺ/ǀƍ0ã ǟ)0,Ę-,&#é ĩ0ƆǺǀƍ, !)=-LtCG KXk`]-<>S bFQ? (öøÂ#:/C =-N`_jG] ȔśêƧǷCÖ8ȕ)° ŽƩƲħ´CŬ ƆǺ*Lt=,? p}LCǪǯäȋóȏĞȀ )Ǒü? žƶ/¯ƪ»Ŀ)0ƴǟǸƶ/ƄŠģȍţ&(»@?*? ƩƲǟ /ÄƿCï =ǎ#ȍC P ȍ ƩƲï-ĵ#ÄƿCƩƲǟ =ǎ#ȍC E ȍ 6#ƩƲǟ-ĵ#ƶ/LjȍC PS ȍ ƩƲ/ï -ĵ#ƶ/LjȍC ES ȍ books.google.co.jp/books?isbn=0123743206