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ƩƲžŷõ ƜȘÝ ¥õȏĞȀ*ȋóȏĞȀ ! #

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Page 1: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004
Page 2: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

1590 Hans Jansen and

Lacharias Jansen

1655 Robert Hooke

1665

Micrographia

Robert Hooke

” Cell ”

! Cell Doctorine 1838

Antonie van Leeuwenhoek

1673 (1677)

1931 Max Knoll Ernst A.F. Ruska

Leeuwenhoek http://teslabar.blogspot.jp/2011/07/blog- post_03.html

Hooke http://

www.pyonta.city.hiroshima.jp/blog/pages/number/268.html

Page 3: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

!!

Page 4: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

10-1 d deci 10-2 c centi 10-3 m milli 10-6 µ micro

10-9 n nano

10-12 p pico

10-15 f femto

10-18 a atto 10-21 z zepto

10-24 y yocto

1024 Y Yotta 1021 Z Zetta 1018 E Exa 1015 P Peta 1012 T Tera 109 G Giga 106 M Mega 103 K Kilo 102 H Hecto 101 D Deca

10-2 c centi10-3 m milli10-6 µ micro10-9 n nano10-12 p pico

Page 5: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

0.1 mm (2~3 µm)

0.2 µm " = 400 nm n sin # = 1.4 0.025 µm

" = 0.004 nm 0.002 nm

, 0.1 nm

Resolution : 2

=0.61 " n sin#

" = n sin # = n =

# =

Fraunhofer W = 0.61 /

Rayleight

http://www.tuat.ac.jp/~katsuaki /el/EL2008/slide_hikari04.pdf

Page 6: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

!! 400~700 nm

!! !! !! !! !! !!

!! 0.004 nm 0.002 nm

!! !!

Page 7: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004
Page 8: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

http://www.isas.jaxa.jp/ISASnews/No.204/ken-kyu.html

Page 9: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

$

Page 10: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

DIC

2 2

DIC2

2

2 a b

Page 11: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

収縮胞 微小管 トリコシスト食胞（DV-1）核 マイオネーム

e-e

Page 12: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

収縮胞 微小管 トリコシスト食胞（DV-1）核 マイオネーム

http://www.nikon-instruments.jp/jpn/PRODUCTS/OPTION/INDEX1.ASPX

Page 13: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

収縮胞 微小管 トリコシスト食胞（DV-1）核 マイオネーム

DAPI: DNA GFP: green fluorescence protein

CFP, YFP: GFP FITC: fluorescein isothiocyanate

Rhodamin: Phalloidin

Alexa: Alexa Fluor

RFP: red fluorescence protein Cy3, 5: cyanine

Page 14: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

Light Amplification by Stimulated Emission of Radiation = = =

http://home.s00.itscom.net/large/ELEC/laser/index.htmlhttp://home.s00.itscom.net/large/ELEC/laser/index.html

http://bioimaging.jp/learn/023/

Page 15: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004
Page 16: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

scaninng

nm

XY Z

nm

Z

http://bioimaging.jp/learn/023/

Page 17: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

DAPI

GFP

Fluorescence in situ Hybridization

Fluorescent Recovery after Photobreach

Page 18: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

~100 kV

~0.004 nm

Page 19: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

= Transmission Electron Microscope (TEM)

Page 20: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

TEM

: OsO4

Quetol 812

80 over night

or 90 nm

2

75 kV

Page 21: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

~10 kV

TEM

= Scanning Electron Microscope (SEM)

Page 22: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

SEM

: OsO4

CO2 t-

5 kV

http://hr-inoue.net/zscience/topics/sem/sem.html

Page 23: © ² ~ wõmail2.nara-edu.ac.jp › ~masaki › CBL-SITE › Other_files › １章...¥õ )0 ¥ 0.2 µm " = 400 nm n sin # = 1.4 ´ h *S m{ ] Ó O) 0.025 µm 6)Î ³ ó )0 " = 0.004

269!

90!

http://www.tech.nagoya-u.ac.jp/event/h23/Vol07/hon_secur/OSEI-1_s.pdf

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P

E

PS ES

books.google.co.jp/books?isbn=0123743206


volume ~ 1 µm x 1 µm x 1 µm

volume ~ 1 µm x 1 µm x 1 µm

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