search.noResults

search.searching

dataCollection.invalidEmail
note.createNoteMessage

search.noResults

search.searching

orderForm.title

orderForm.productCode
orderForm.description
orderForm.quantity
orderForm.itemPrice
orderForm.price
orderForm.totalPrice
orderForm.deliveryDetails.billingAddress
orderForm.deliveryDetails.deliveryAddress
orderForm.noItems
Microscopy Pioneers


method into a routine technique [ 16 , 17 ] that can now be quickly learned and applied to a wide range of biological applications. Nevertheless, the foundations of this success were the two beautiful papers by Tokuyasu in 1973 and 1978 [ 2 , 14 ].


Kiyoteru Tokuyasu Aff ectionately called Tok, he was born in Nagasaki, Japan, in 1925. He studied physics (with a degree 1949) and completed a PhD in medical sciences at Kyushu University in 1957. His PhD work focused on ultrastructure of the cat retina [ 18 ] (Tokuyasu and Yamada 1959). Aſt er two years as associate professor in the electron microscopy lab of Kurume Medical College, he was hired as head EM designer for Hitachi (1958–1963). He moved to the USA in 1964 as associate research biologist in the Department of Pathology at UCLA. In 1969 he was attracted to UCSD by Jon Singer, who had the vision to realize that the cryo-sectioning method had great potential for immunolabeling and the even greater vision to give Tok the support he needed to create the Tokuyasu method. Tok was a regular and welcome visitor to Europe. He was for many years an instructor in the annual European Molecular Biology Organization (EMBO) practical course. It is remarkable to note that the fi rst course organized in 1979, at the European Molecular Biology Laboratory in Heidelberg, Germany, focused exclusively on the Tokuyasu method described in an article that was published only a year earlier! T is was, in retrospect, highly ambitious because in those days the ultramicrotomes were rudimentary, and the technique required much patience. Today, the technique can be rapidly learned and is taught in several specialized courses. Prof. Tokuyasu was awarded the MSA Distinguished Scientist Award in 2000.


Courses that Teach the Tokuyasu Cryosectioning Method T e fastest way to learn about immunocytochemistry and how to prepare thawed cryosections for immunolabeling is to attend a course. T ese courses are off ered occasionally by various organizations. T e European Molecular Biology Organization (EMBO) sponsors an annual “Advanced Electron Microscopy for Cell Biology” practical course. T e course teaches the Tokuyasu technique for immunolabeling as well as covering all aspects of immunocytochemistry for electron microscopy and other important EM specimen preparation methods for biologists. In 2018 the course was held at the University


of Würzburg, Germany. In 2019 the course will be held at the University of South Bohemia in the Czech Republic. T e Department of Cell Biology at the University of Utrecht holds basic and advanced courses for sectioning and immunolabeling ( http://www.cellbiology-utrecht.nl/em-courses.html ). Courses held in response to demand are also organized by Marine Reef International ( http://www.marinereef.com/courses.php ) and RMC-Boeckeler ( http://www.rmcboeckeler.com ). Electron Microscopy Sciences include cryosectioning of fi xed frozen biological material in their academy curriculum ( https://www. emsdiasum.com/microscopy/academy/courses/courses.aspx ). Cryoultramicrotomes and information about them can be obtained from Leica Microsystems and RMC-Boeckeler.


References [1] H Fernandez-Moran and AO Dahl , Science ( 1952 ) 116 ( 3018 ) 465 – 67 .


[2] K Tokuyasu , J Cell Biol 57 ( 2 ) ( 1973 ) 551 – 65 . [3] J Dubochet and A McDowall , J Microsc Oxford 124 ( 3 ) ( 1981 ) 3 – 4 .


[4] P Brüggeller and E Mayer , Nature 288 ( 5791 ) ( 1980 ) 569 – 71 . [5] G Griffi ths et al. , J Ultra Mol Struct R 89 ( 1 ) ( 1984 ) 65 – 78 . [6] JW Slot and HJ Geuze , Nat Protoc 2 ( 10 ) ( 2007 ) 2480 – 91 . [7] K Tokuyasu and S Okamura , J Biophys Biochem Cy ( 1985 ) 305 – 08 .


[8] A Christensen , A way to prepare frozen thin sections of fresh tissue for electron microscopy and in Autoradiography of Diff usible Substances eds LJ Roth and WE Stumpf, Academic Press , London, New York , 1969 , pp. 349 – 62 .


[9] AK Christensen , J Cell Biol 51 ( 3 ) ( 1971 ) 772 – 804 . [10] J McLean and S Singer , P Natl Acad Sci 65 ( 1 ) ( 1970 ) 122 – 28 . [11] WP Faulk and GM Taylor , Immunochemistry 8 ( 11 ) ( 1971 ) 1081 – 83 .


[12] MJ Horisberger et al ., Experientia 31 ( 10 ) ( 1975 ) 1147 – 49 . [13] K Tokuyasu and S Singer , J Cell Biol 71 ( 3 ) ( 1976 ) 894 – 906 . [14] K Tokuyasu , J Ultra Mol Struct R 63 ( 3 ) ( 1978 ) 287 – 307 . [15] LY Bourguignon et al ., J Cell Physiol 95 ( 3 ) 1978 ) 239 – 57 . [16] E Bos et al ., J Struct Biol 175 ( 1 ) ( 2011 ) 62 – 72 . [17] P Webster and A Webster, “Cryosectioning Fixed and Cryoprotected Biological Material for Immunocytochemistry ,” in Electron Microscopy , ed J Kuo, Humana Press , Totowa, NJ , 2014 , pp. 273 – 313 . [18] K Tokuyasu and E Yamada , J Cell Biol 6 ( 2 ) ( 1959 ) 225 – 30 .


2018 July • www.microscopy-today.com


47


Page 1  |  Page 2  |  Page 3  |  Page 4  |  Page 5  |  Page 6  |  Page 7  |  Page 8  |  Page 9  |  Page 10  |  Page 11  |  Page 12  |  Page 13  |  Page 14  |  Page 15  |  Page 16  |  Page 17  |  Page 18  |  Page 19  |  Page 20  |  Page 21  |  Page 22  |  Page 23  |  Page 24  |  Page 25  |  Page 26  |  Page 27  |  Page 28  |  Page 29  |  Page 30  |  Page 31  |  Page 32  |  Page 33  |  Page 34  |  Page 35  |  Page 36  |  Page 37  |  Page 38  |  Page 39  |  Page 40  |  Page 41  |  Page 42  |  Page 43  |  Page 44  |  Page 45  |  Page 46  |  Page 47  |  Page 48  |  Page 49  |  Page 50  |  Page 51  |  Page 52  |  Page 53  |  Page 54  |  Page 55  |  Page 56  |  Page 57  |  Page 58  |  Page 59  |  Page 60  |  Page 61  |  Page 62  |  Page 63  |  Page 64  |  Page 65  |  Page 66  |  Page 67  |  Page 68  |  Page 69  |  Page 70  |  Page 71  |  Page 72  |  Page 73  |  Page 74  |  Page 75  |  Page 76