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ANNUAL REPORT APRIL 1999 TO MARCH 2000 ELECTRON MICROSCOPY UNIT FACULTY OF MEDICINE NATIONAL UNIVERSITY OF SINGAPORE ANNUAL REPORT APRIL 1999 TO MARCH.

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Presentation on theme: "ANNUAL REPORT APRIL 1999 TO MARCH 2000 ELECTRON MICROSCOPY UNIT FACULTY OF MEDICINE NATIONAL UNIVERSITY OF SINGAPORE ANNUAL REPORT APRIL 1999 TO MARCH."— Presentation transcript:

1 ANNUAL REPORT APRIL 1999 TO MARCH 2000 ELECTRON MICROSCOPY UNIT FACULTY OF MEDICINE NATIONAL UNIVERSITY OF SINGAPORE ANNUAL REPORT APRIL 1999 TO MARCH 2000 ELECTRON MICROSCOPY UNIT FACULTY OF MEDICINE NATIONAL UNIVERSITY OF SINGAPORE

2 Contents Introduction Staff of the EM Unit Summary Specimen Processed Users Revenue Activities Visitors On-going Activities

3 Introduction

4 INTRODUCTION Over the twelve-month period, the Electron Microscopy (EM) Unit in the Faculty of Medicine consolidated on the initiatives from the previous years. The Unit continued to conduct specialised training courses in conjunction with FEI/Philips Electron Optics on Basic Scanning Electron Microscopy and Field Emission Gun Scanning Electron Microscopy. This year we saw many participants from the industries. In addition, an Engineers Technical Training course was initiated using the Units 208S Transmission Electron Microscope. The facilities continued to be used by various vendors, as a reference site and for demonstrations on the instruments, throughout the year for their potential customers. Several foreign visitors also visited our Unit to look at our facility and organisation, with the intention of forming some form of collaboration with us. Usage of the Units facility was maintained among the Faculty users. However, during this period there was a substantial increase in usage by non-faculty users (details in report). One major activity that the Unit is very much involved is in the organisation of the 7 th Asia-Pacific Electron Microscopy (APEM) conference (26 to 30 June, 2000, Singapore). The effort seems to have paid off as the response for this conference is highly encouraging with an expected participation of about 500 delegates. Most of the pre-conference workshops will take place at the EM Unit.

5 The other major project that the staff of the Unit are concentrating on is to produce a handbook on Environmental Scanning Electron Microscopy in Life Sciences for the Novice. The aim is to launch this book at the 7 th APEM conference. It is hoped that the tips provided would assist users of this technology in Life Sciences. As usual, the EM Unit would not be able to maintain its level of activity without the usual strong support and cooperation from our users and collaborators, including our commercial partners. The Unit is very grateful for this continuous support. The efforts and the contributions of the central technologists are also appreciated. Mary Ng Mah Lee Head, Electron Microscopy Unit Faculty of Medicine e-mail: micngml@nus.edu.sgmicngml@nus.edu.sg

6 Staff

7 Staff of the EM Unit From left to right : (top) Patricia, Suat Hoon, Catherine, Prof Mary, (bottom) Cynthia, Micky. During this year, the Unit saw the departure of Mr. Ng Chee Peng. Ms Cynthia Ng was subsequently recruited and the staff number was maintained at five. As mentioned previously, their main duties are to keep the instruments of the Unit in good running order. In addition, they provide in-house training courses for new users who want to use the Units facilities. The staff members have been kept rather busy with gathering good images for the Handbook on Environmental Scanning Electron Microscopy. Hopefully, this book will be a success for the Unit as it is our first attempt at such activity. Responsibilities for the instruments are divided up as follows : Miss Tan Suat HoonCM 120 BioTwinTEM/STEM/Cryo/EDX JEM 1220 TEM/EDX Cryo-techniques Miss Catherine TangFreeze Fracture/etching ESEM SEM specimen preparation

8 Amongst all their routine duties, when time permits these technologists also take on small contract projects from the hospitals, other faculties and institutions. These contracts are revenue generating for the Unit. Miss Patricia NettoFEG-SEM/EDX Image analyser and processor Digital imaging Ms Cynthia Ng208S TEM 1200EX TEM Cryo-techniques Miss Micky Leong JSM 5600 LV SEM Freeze Fracture/etching TEM and SEM specimen processing

9 Summary

10 Since we have entered into the 21 st century, it might be best to summarise the acquisition of instruments of the EM Unit during the last few years of the 20 th century. As mentioned previously, the Unit had a major upgrading exercise together with an expansion of space in 1996. From July 1996 to March 1999, the following instruments were purchased: TEM CM 120 BioTwin with EDX Freeze Fracture Replica and Freeze Etching Image Analysis and Processing System with full range of Digital Capture System Field Emission Gun (FEG) Scanning Electron Microscope with EDX 208S Transmission Electron Microscope JEM 1220 with ISIS EDX System Low Vacuum Scanning Electron Microscope (LV-SEM) Environmental Scanning Electron Microscope (ESEM) Critical Point Dryer for SEM specimen processing High Resolution Coating System for FEG and STEM specimens Two Ultramicrotomes with CCD attachments Magnetic Field Cancellation Devices costing S$13, 000 each Lab-Top Computer

11 Some new accessories have also been added to our existing microscopes. These were purchased from the revenue generated from the activities of the EM Unit. The total sum was SIN$ 55,000. 1) Operation keyboard for the JSM 5600LV that is able to drive the full operation functions of the microscope. We found the mouse operation not responsive enough for some of our users. 2) 1440dpi photo quality, high resolution printer linked to the JSM 5600LV, to allow users to get good quality hard copies of their images immediately without using the usual roll of film. 3) Scaler and calibration software for JSM 5600LV users to measure structures directly on the image on screen. 4) Image expansion memory for high resolution imaging on the JSM 5600LV. 5) Scan rotation on the JSM 5600LV for electronic rotation of the image of the structure during viewing, because orientation is important for some images

12 6)Permanent storage specimen cartridge base, for the JEM 1220, that can be interchanged, so that important grids can be locked in position. If this grid needs to be viewed again, a particular point of the grid can be easily located. 7)Full keyboard for text linked to the JEM 1220 microscope for typing text on the negatives 8)Beryllium analytical retainer is use for elemental analyses (EDX) as it gives low background compared to standard holder. Low background is needed for good elemental profile of the specimen (JEM 1220 TEM) 9)EDX kit -this is a software for the EDX for JEM 1220 TEM 10)SEMI-STEM attachment expand the functions of the EDX system by providing the facility for doing elemental mapping on the image of the specimen. This is a much cheaper option to a full STEM attachment. This was the first Unit to be installed in Singapore at the time we purchase it (JEM 1220 TEM). For April 1999 to March 2000, the following new instruments and accessories were purchased. All the items were purchased using the revenue generated by the activities of the EM Unit. Hydrophilic Glow Discharger Hydrophobic Glow Discharger High Resolution Photomonitor for the ESEM These purchases came to a total of SIN$ 32,815.13

13 Specimen Processed

14 NUMBER OF SPECIMENS PROCESSED July 1996 to March 1999 TEM = 3261 SEM = 1013 Cryo- techniques = 334 Freeze fracture replica = 131 EDX analysis = 170 Image Analysis = 503 Slides/transparencies/presentation media = 2387 April 1999 to March 2000 TEM = 924 SEM = 431 Cryo- techniques = 80 Freeze fracture replica = 20 EDX analysis = 200 Image Analysis = 672 Slides/transparencies/presentation media =1228

15 Users

16 April 1999 to March 2000 – the list is enclosed below. I. National University of Singapore, Faculty of Medicine Anatomy Department Staff :Bay Boon Huat Charanjit Kaur Gopalakrishnakone P Leong Seng Kee Ling Eng Ang Technical Staff :Yick Tuck YongNg Leh See Chan Yee Gek Research Assistant : Liu HaipingZ Emmanuel Selvanayagam Qiang Mei LIST OF USERS

17 Students : Anita Selvakumari Jayasurya Aye Mya Mya Cao QiongGao Rong Jin RongXianK N Srinivasan Keiko ObaroLi JingSong Lu Xin RongMyint Swe Ng Chee HonQi Long R Ashok BalajiS Dinesh KumarSelvanayagamNirthanan T ShobaTang Fengru Xie Jian PingYou Yimei Wang DongWang Xinsheng Wang XuWu Ya Jun Zeng XiaoXiaZhang Li

18 Biochemistry Department Staff :Chang Chan FongKandiah Jeyaseelan John Candlish Technical Staff :Lalana Youngpritisporn Student :Khoo Keng MArunmozhiararsi Chua Kim Lee Fatemeh Aririyan Microbiology Department Staff :Chan Soh HaNg Mah Lee Ho BowSong Keang Peng Kevin TanVincent Chow Lee Yuan KunYap Eu Hian Mulkit Singh Technical Staff :Ho Lip ChuenLoy Boon Pheng Josephine HowePhoon Meng Chee

19 Students : A M A Nasirudeen Tay Wee Lee, Jonathan Eugene Quek Se Thoe Su Yun Joogal Sudhir Kumar Chandromohan Lim Chai Yen Wu Jack Han, Victor Lim Li Lian Wee Boon Yu Lim Hui Ying Yap Ann Teck Loke Shee Mei Raudhhah Abdul Rahim

20 Obstetrics & Gynaecology Department Staff :Ratnam S S Technical Staff : Jean Ho Opthalmology Department Staff :The Late Chew Sek JinLim S M, Arthur Research Staff : Hu Yang Orthopaedic Surgery Department Staff :John ChiaPho W H P ThiagarajanV Prem Kumar Technical Staff : Hersharan Kaur d/o Balder SYong S C Research Assistant :Kenjiko HasegawaZhu Hong Jiang HuaBarry P Pereira Student : Chan Wai Kam

21 Otolaryngology Department Staff :Ruan Runsheng Paediatrics Department Staff :Lee Bee Wah Wai Cheung Chua Kaw Yan Pathology Department Staff :Aileen WeeShanmugaratnam K Lee Hin PengSinniah R Lee Yock SunTeh Ming Technical Staff : John Chia Lam San

22 Pharmacology Department Staff :Benny TanSim Meng Hwoon Peter Wong Technical Staff : Tang Fengru Research Assistant : Durairaj Ponraj Student :Loo Li ShenWu Xiao Hua Physiology Department Staff :Koh D RShazib Pervaiz Technical Staff : Ho Chiu Han Research Assistant : S Mohammed Ali

23 Department of Surgery Staff :James TanTan Tiang Lee, Walter Kewawan Esuwaranathan Timothy Lee Lim Thiam Chye Research Assistant : K M Ameer AliZhang Li Student : Achuth H NCarmel Lawrencia

24 II. National University of Singapore, Non-Faculty of Medicine Faculty of Dentistry Preventive Dentistry Department Staff :Hsu Chin-Ying Stephen Research Assistant : Shi Hong Restorative Dentistry Department Staff :Thean Pik Yen, Hilary Faculty of Engineering Chemical Engineering Department Staff :Ting Yen PengTeo Wah Koon Research Assistant : Atul Kumar MittalSun Gang Student : Yang Ping

25 Faculty of Science Biological Sciences Department Staff :Hugh Tan Tiang Wah Student : Goh Wee Kee Pauline Tan Wei Leng Stephanie Seah Chemistry Department Staff :J D RanfordLi Fong Yau, Sam Physics Department Staff :Frank Watt

26 III. Other Institutes / Industries and Hospitals Defence Medical Research Institute Staff :S M Moochnala Institute of Molecular and Cell Biology Staff :Heinz Horstmann Xu Jing Institute of Molecular Agrobiology Staff :Megan GriffithTerje Dokland Myong Ok KwonYin Zhan Sundaresan, VenkatesanWilliam Chia Wang Yue National Cancer Centre Staff :Hui Kam Man Wu XiaFang

27 National University Medical Institutes Confocal Microscopy Lab & Flow Cytometry Lab Research Scientist : Hanry Yu Research Officer : Xu Xi Lab Technologists : Er Poh Nee, ConnieNg Bee Ling Student : Chia Ser Mien Singapore Eye Research Institute Staff :Hu Yang Wilson Wong Singapore Polytechnic Staff :Anna Yeo Tan Tock Seng Hospital Staff :Fong Sean Yean Low Chee Kuang Jeffery Chew

28 Singapore General Hospital Staff : Robert NgFei Xun,Beatty Users from Industries who are customers of Philips Singapore Pte Ltd 3M Adaptec Manufacturing (Singapore) Pte Ltd Chartered Semiconductor Manufacturing Ltd Hewlett-Packard Singapore Quantum VTM Users from Industries who are customers of JEOL Asia Pte Ltd Glaxo Wellcome

29 Revenue

30 REVENUE GENERATED July 1996 to March 1999 Fund from usage of equipment S$ 157,600 Fund from contract work S$ 47,300 Fund from workshops S$ 12,091 Consumable S$ 34,900 Total income S$ 247,891 April 1999 to March 2000 Fund from usage of equipment S$ 92,676 Fund from contract work S$ 13,343 Fund from workshops S$ 5,400 Consumable S$ 14,305 Total income S$ 125,724

31 Activities

32 ACTIVITIES a) Publications/conference papers/dissertations/theses generated from the use of the EM Unit's facility July 1996 to March 1999 The details on the publications have been given in previous annual reports. A summary of the total numbers for this period is given below. International Refereed Journals = 76 Conference Papers = 75 Ph.D. Theses = 12 MSc Theses = 8 Honours Dissertation = 4 UROP Projects = 4 Advance Diploma = 1 April 1999 to March 2000 International Refereed Journals = 42 Conference Papers = 30 Ph.D. Theses = 4 MSc Theses= 1 Honours Dissertation = 3 UROP Projects = 1

33 April 1999 to March 2000 I. INTERNATIONAL REFERENCE JOURNALS 1.F.R. Tang, M.K. Sim. Pre- and /or post-synaptic localisation of metabotropic glutamate receptor 1 BX (mGluR1 ) and 2/3 (mGluR2/3) in the rat spinal cord., Neuroscience Research 34 (1999) 73-78 2.Tan, Y, R Sinniah, B H Bay and G Singh, Metallothionein expression and nuclear size in benign borderline and malignant serous ovarian tumors. Journal of Pathology 189 (1999, United Kingdom) – 60-65. 3.Sinniah R and T N Khan, Ultrastructural study of renal tubular basement membrane in tubulointerstitial damage. Ultrastructural Pathology (1999, Scandinavia) 4. Tan Yi, Sinniah R, Bay B H, Singh G, Expression of Metallothionien and Nuclear Size in discrimination of malignancy in musicinous ovarian tumors. International Journal of Gvneological Pathology (1999,USA) 5. Henkel AW, Meiri H, Horstmann H, Lindau M, Almers W. Vesicles open and close rhythmically during constitutive exo-and endocytosis in chromaffin cells. EMBO J 2000 Jan 4;19(1) : 84-93 6.Ong, W Y and K Mackie, A light and electron microscopic study of the CB1 cannabinoid recepter in the primate spinal cord. Journal of Neurocytology, 27 (1999) : 719-730. (United States). 7.Ong, W Y and K Mackie, A light and electron microscopic study of the CB1 cannabinoid receptor in primate brain. Neuroscience, 92 (1999) : 1177 - 1191. (United States). 8.Ong, W Y, Y P Soh, C Y Hu, P G Pentchev and S C Patel, Neuronal localization of sterol regulatory element binding protein-1 (SREBP-1) in the rodent and primate brain : A light and electron microscopic immunocytochemical study. Neuroscience (2000) 9.Kumar, S.D. and Tay SSW (1999) Ultrastructural changes in the atrioventricular valves of streptozotocin - induced diabetic rats. Journal of Brain Research (Germany) Vol. 39 : 557-564. 10.V Zaman, J Howe, M Ng and TK Goh,Ultrastructure of the Endolimax nana cyst Parasitology Research 86: 54- 56 (2000)

34 11.PY Zeng, J Hua, HC Ng and B. Ho, Unchange characteristics of Helicobacter pylori during its morphological conversion Microbios 98: 51-64, 1999 12.V Zaman, M Zak. M Manzoor, J Howe, M Ng, Postcystic development of Blastocystis hominis, Parasitology Research 85: 437-440, 1999 13.Zaman, V, M Zakai, J L C Howe, M M L Ng, D D Leipe, JD Silberman and M Sogin, Hyperamoeba isolated from human feces : Description and Phylogenetic Affinity. European Journal of Protistology, 35 (21 June 1999) : 197-207. (Germany). 14.Lim, HY and M M L Ng, A different mode of entry by dengue-2 ventralisation escape mutant virus. Archives of Virology, 144 (1999) : 989-995. (Austria). 15.Zaman, V, J L C Howe, M M L Ng and T K Goh, Scanning electron microscopy of the surface coat of Blastocystic hominis. Parasitology Research, 85 (1999) : 974-976. (Austria) 16.Zaman, V, J L C Howe and M M L Ng, Post- cystic development of Blastocystis hominis. Parasitology Research, 85 (1999) : 437-440. (Austria). 17.Zaman, V, J L C Howe, M M L Ng and T K Goh, Ultrastructure of Endolimax nama cyst. Parasitology Research, 86 (January 2000) : 54-56. (Austria). 18.Sudhir, K and M M L Ng, Mycovirus from Shiitake mushroom and Cancer-Prevention. The Singapore Biochemist, Vol 12, edited by SI Koh, WY Kwok and HE Khoo. Singapore : Singapore Society for Biochemistry and Molecular Biology, March 1999. 6 pp. 19.Gong, NL, Armugam, A. and Jeyaseelan, K (1999) Postsynaptic short neurotoxins from Pseudonaja textiles : cDNA cloning, expression and protein characterization. Eur J Biochem. 265: 982-989. 20.Lachumanan R., Armugam, A., Ponraj, D., Gopalakrishnakone, P., Tan, CH and Jeyaseelan, K(1999) In situ hybridization and immunohistochemical analysis of the expression of cardiotoxin and neurotoxin genes in Naja naja sputatrix. J. Histochem. Cytochem. 47, 551-560

35 II. PUBLICATIONS IN PRESS – INTERNATIONAL REFERENCE JOURNALS 1.Jayasurya A, Bay BH, Yap WM, Tan NG. Correlation of metallothionein and apoptosis in nasopharyngeal carcinoma. British Journal of Cancer, 1999. 2.Subramaniam N., Lohe E., Horstmann H., Griffiths G., Habermann A. and Wanjing Hong, Preferential association of syntaxin 8 with early endosomes, Journal of Cell Science 2000 in press 3.Qi Zheng, Xiaoning Si, H. Horstmann, Yue Xu, Wanjin Hong and Catherine J. Pallen Prenylation-dependent Association of Protein Tyrosine Phosphatases PRL-1, -2, And 3 with the Plasma Membrane and the Early Endosome, Journal of Biological Chemistry, submitted 4.Menno L.W. Knetsch; Nicole Schaefer; Heinz Horstmann; Dietmar Manstein Racin, a multifunctional rac-regulator in Dictyostelium discoideum, J Cell Biol 1999 submitted 5.Ms Mi Wei Guan (MSC), Development of Immunodiagnostics for the detection of Burkholderia pseudomallei, the camature agent of mehadosin.. Submitted 6 th Nov 1999 6.Professor Ng Soon Chye, Light microscopic and ultrastructural observations of enzymatically treated human blastocysts : zona-free blastocyst transfer and rescue of blastocysts with hatching difficulties (in preparation, to be submitted to Human Reproduction) 7.Li, J., Foo, M.L., Wang, Y., Ng, H.T., Jaenicke, S., Xu, G.X., Li, S.F.Y. The Synthesis of Single-Walled Carbon Nanotubes by CVD Catalyzed with MCM-41 Powder. Science and Application of Nanotubes (Tomanek, D. and Enbody, R., Kluwer Academic Publishers) in press. 8.Ng, H.T., Fang, A.P., Chan, L. and Li, S.F.Y. Neighbouring-seeding-mediated highly selective electroless copper metallization on non-catalytic surfaces.

36 9.Fang, A.P., Ng, H.T., Su, X.D., Li, S.F.Y. Soft lithography-mediated submicrometer patterning of self- assembled monolayer of hemoglobin on ITO surfaces. Submitted for publication. 10.Zaman, V, J L C Howe and M M L Ng, Scanning electron microscopy of Chilomastix mesnili : (Wenyon, 1910) A lexieieff, 1912. Parasitology Research (2000). (Austria). 11.Se Thoe, SY, AE Ling and M M L Ng, Alteration of virus entry mode a neutralisation mechanism for dengue-2 virus. Journal of Medical Virology (2000). (Austria) 12.Jeyaseelan, K., Armugam, A., Ma, DH and Tan, NH.(2000). Anticoagulant Group I phospholipase A genes of Naja naja sputatrix. Molecular Biology and Evolution. 13.M. M. Aye, G. Singh and S.S.W. Tay Ultrastructural change in the Sinuatrial and Atrioventricular Nodes of the Heart of the Monkey (Macaca Fascicularis) after bilaterial vagotomy (Journal of Brain Research) 14.M. M. Aye, G. Singh and S.S.W. Tay Ultrastructural study of the effects of 6-hydroxydopamine on theSinuatrial and Atrioventricular Nodes of the Heart of the Monkey (Macaca Fascicularis) (Journal of Brain Research) 15.M. M. Aye, G. Singh and S.S.W. Tay An ultrastructural study of the Innervation of the Heart of the Monkey (Macaca Fascicularis) (Annals of Microscopy). 16.Farooqui, A A, W Y Ong and L A Horrocks, Brain cytosolic phospholipase A 2 : Immunocytochemical localization, properties roles and involvement in neurological diseases. Neuroscientist (2000). (United States) Publication no : 0104821 17.Ng, C H and W Y Ong, A light and electron microscopic study of GAT- 1 positive cells in the monkey brainstem and spinal cord. Journal of Brain Research (2000). Publication no : 0117619 18.Wang, X S and W Y Ong, A light and electron microscopic study of GAT-1 in the monkey basal ganglia. Journal of Neurocytology (2000). Publication no : 0117620

37 19.Lu, X R, W Y Ong and K Mackie, A light and electron microscopic study of the CB1 Cannabinoid recepter in monkey basal forebrain. Journal of Neurocytology (2000). Publication no : 0117621 20.Ong, W Y, C Y Hu, O P Hjelle, O P Ottersen and B Halliwell, Changes in glutathione in the hippocampus of rats injected with kainate - depletion in neurons and upregulation in glia. Experimental Brain Research (2000). Publication no : 0117622 21.Ong, W Y, H M Lim, T M Lim and B Lutz, Kainate-induced neuruonal injury leads to persistent phosphorylation of CREB in glial and endothelial cells in the hippocampus. Experimental Brain Research (2000). Publication no : 0117624 22.Ong, W Y, Y P Soh, C Y Hu, P G Pentchev and S C Patel, Neuronal localization of sterol regulatory element binding protein-1 (SREBP-1) in the rodent and primate brain : A light and electron microscopic immunocytochemical study. Neuroscience (2000) Publication no : 0117625

38 III. CONFERENCE PAPERS 1.H. Horstmann, Zhang Tao and Wanjin Hong, Electron Microscopic Localization Of the Intermediate Compartment in VSV infected Unfixed Rapid Frozen Normal Rat Kidney Cells (NRK) Abstract : International Meeting on Transport of proteins and Membranes in Eukaryotic Cells, Goettingen, 1999, September 29 2.ML Ng, SH Tan, LN Leong, EK Tang, P Netto and J Howe, Environmental Scanning Electron Microscopy Technology for Life Sciences, 17 th Conference of the Electron Microscopy Society of Thailand, 1999 3.ML Ng, SH Tan, LN Leong, EK Tang, P Netto and J Howe, Is Envirmental Scanning Electron Microscopy Suitable for Biological Specimens? 8 th Scientific Conference of the Elecron Microscopy Society Malaysia, Genting Highlands, Malaysia 2-4 Dec 1999 4.Sinniah, R. Mechanisms of renal injury in malaria infection.(Abstract pg70), I in.Symposium. Malaria : New Insight into an old World-wide disease. 3 rd National University of Singapore - National University Hospital Annual Scientific Meeting. 28 & 29 th August 1999, Singapore. 5.R. Ashok Balaji, Gopalakrishnakone. P. " Natural toxins from Marine Conus Snail - A Review" Oral presentation, 10 th Asia pacific Military Medicine Conference, 7-12 May 2000, Singapore. 6.R. Ashok Balaji, Gopalakrishnakone.P, Bay Boon Huat,. Kazuki Sato. The structural characterization of venom apparatus and radula tooth of Indo-pacific molluscivorous Gastropod (Cone Snail) species Conus amadis. Abstract, June 2000, 7 th Asia-Pacific Electron Microscopy Conference Singapore. 7.R.Ashok Balaji, Toru Sasaki, Gopalakrishnakone. P, Kazuki Sato, Bay Boon Huat. Toxin myopathy induced by a toxin (ST28 or Covalitoxin-I) and crude venom obtained from Coremiocnemius validus. Abstract, Aug 3 rd NUH-Faculty of Medicine Annual Scientific Meeting 1999. Singapore. 8.Ng, H.T., Foo, M.L., Lim, R., Li, J. and Li, S.F.Y. Direct Controllable Non-Photolithographic Patterning and CVD Growth of 2-Dimensional Arrays and Complex Intertwined Network of Carbon Nauotubes. Submitted to The International Workshop on Advances in Materials Science and Technology. 9.Fang, A.P., Ng, H.t., Su, X.D. and Li, S.F.Y. Fabrication of miniaturized biosensors via soft lithography- mediated self-assembled monolayer of hemoglobin on ITO surfaces. Submitted to The International Workshop on Advances in Materials Science and Technology.

39 10.Kevin SW Tan, J Howe, GC Ng, NP Ramachandran, A M A Nasirudeen, EH Yap, M Singh. Programmed cell death in colony forms of Blastocystis hominis in Microbes and diseases: cause, effect and pharmacotherapy. Annual Scientific meeeting of Biomedical Research and Experimental Therapeutics Society of Singapore/Singapore Society of Biochemisry and Molecular Biology/ Singapore Society of Microbiology and Biotechnology. National University of Singapore, 13-14 August 1999 11.AMA Nasirudeen, Kevin SW Tan, EH Yap, M Singh. Programmed cell death in Blastocystis hominis after exposure to a cytotoxic monoclonal antibody. Annual Scientific meeting of Biomedical Research and Experimental Therapeutics Society of Singapore/Singapore Society of Biochemisry and Molecular Biology/ Singapore Society of Microbiology and Biotechnology. National University of Singapore 13-14 August 1999 12.AMA Nasirudeen, Eugene Quek, EH Yap, M Singh and Kevin SW Tan. Apoptosis-like death in Blastocystis hominis induced by colony growth and cytotoxic monoclonal antibody. Asia-Pacific Congress on the Biology of the Environment. National University of Singapore 21-24 November 1999 13.MC Phoon, J Howe, V Chow TK, Charles DesBordes. Linoleic and Linolelaidic acids affect the proliferation and ultrastructure of Molt-4 lymphoblastic leukemai cells. 24 th World Conference of Medical Technology Vancouver, Canada. 8-23 June 2000 (accepted for presentation) 14. J Howe, XQ Chen, M singh, LC Ho, SW Tan and EH Yap. Ultrastructural changes during in vitro excystation of Blastocystis Ratti, 24 th World Conference of Medical Techology. Vancouver, Canada 18-23 June 2000 (accepted for presentation) 15.Wang Woo, YH and M M L Ng, Randomized Clinical Trial to Evaluate the Medical Effects of Shiitake Mushrooms and Products. Paper presented at Federation of Asia-Pacific Microbiology Societies "Microbiology and Infectious Diseases in the Asia Pacific : The 21st Century, 24-27 May 1998, Suntec City, Singapore. 16.Lok, SM, M M L Ng and J Aaskov, Identification of functional epitopes on the e-protein of dengue 2 virus (Abstract). In Abstracts of the XIth International Congress of Virology, compiled by Organising committee, pp. 328. Australia : Australian Microbiology Society, August 1999. (Paper presentated at XI International Congress of Virology, 9-13 August 1999, Sydney Convention Centre, Darling Harbour, Australia).

40 17.Se Thoe, SY, AE Ling and M M L Ng, Inhibition of normal entry pathway for Dengue type 2 virus by monoclonal antibody (Abstruct). In Abstracts of the XIth International Congress of Virology, compiled by Organising Committee, pp. 11. Australia : Australian Microbiology Society, August 1999. (Paper presented at XIth International Congress of Virology, 9-13 August 1999, Sydney Convention Centre, Darling Harbour/Sydney, Australia). 18.Lim, HY and M M L Ng, Entry and Replication of Dengue Virus in Baby Hamster Kidney Cells. Third Asia Pacific Organisation of cell Biology Congress, 24-28 August 1998, Senri Life Science Centre, Osaka, Japan. 19.Lok, SM and M M L Ng, Comparison of Amino Acid Changes in the Envelope protein of Dengue 2 virus (New Guinea C) Grown in C6/36 (Aedes albopictus) and Baby Hamster Kidney (BHK) Cells (Abstracts). Third Asia Pacific Organisation of Cell Biology Congress, 24-28 August 1998, Senri Life Science Centre, Osaka, Japan. 20.Kumar, S and M M L Ng, Interferon Induction : A Mechanism to Explain Antitumour Activity and Defective AKT Retrovirus Production By Lentinus edodes Extract (Abstract). Fourteenth International Congress on Electron Microscopy, 31 August - 4 September 1998, Cancun Convention Centre, cancun, Mexico. 21.Tan, SH and M M L Ng, Close Association of Flavivirus Envelope Protein with Nucleocapsids During Transportation to Cell Surface (Abstract). Fourteenth International Congress on Electron Microscopy, 31 August - 4 September 1998, Cancun Convention Centre, cancun, Mexico. 22.Lok, SM and M M L Ng, Fluorescent Patterns of Dengue 2 Neutralisation Resistant Virus Obtained from Confocal Microscopy (Abstract). Fourteenth International Congress on Electron Microscopy, 31 August - 4 September 1998, Cancun Convention Centre, cancun, Mexico. 23.Yap, AT, S Kumar and M M L Ng, Immunopotentiation by Constituents of Shiitake Mushroom (Lentinus edodes) : Role of Lentinan Extracted By Novel Method (Abstract). International Symposium Science and Cultivation of Mushrooms, 12-15 October 1998, Nanjing International Conference Hotel, Nanjin, China. 24.Wang Woo, YH, S Kumar and M M L Ng, Immunomodulation and Alleviation of Respiratory Symptoms by Shiitake Mushrooms. International Symposium on Science and Cultivation of Mushrooms, 12-15 October 1998, Nanjing International Conference Hotel, Nanjin, China.

41 25.Kumar, S and M M L Ng, Immunopotentiation by Constituents of Shiitake Mushrooms (Lentinus edodes) : Role of Virus-Like Particle. International Symposium on Science and Cultivation of Mushrooms, 12-15 October 1998, Nanjing International Conference Hotel, Nanjin, China. 26.Yap, AT, K Sudhir and M M L Ng, Partially purified Lentinan from Shiitake mushroom (Lentinus edodes) still retain antitumor activity. In Mushroom Conference 1999 (CD ROM), edited by Andrew Broderick and Tan Nair, pp. 215- 224. Australia : The Horticultural Research and Development Corporation, Australia, October 1999. (Paper presented at 3 rd International Conference on Mushroom Biology and Mushroom products, 12-16 December 1999, Novotel, Brighton/Sydney, Australia). 27.Sudhir, K and M M L Ng, Apoptosis and cytokine induction studies by virus-like particles from Lentinus edodes (Shiitake) mushroom on murine lymphoma. In Mushroom Conference 1999(CD ROM), edited by Andrew Broderick and Tan Nair, pp. 167-174. Australia : The Horticultural Research and Development Corporation, Australia, October 1999. (Paper presented at 3 rd International Conference on Mushroom Biology and Mushroom products, 12- 16 December 1999, Novotel, Brighton/Sydney, Australia). 28.Ng, M M L, What is a cell? Cell Biology International (Paper presented at Third Asia-Pacific Organisation of cell Biology, 24-28 August 1998, Senri Life Science Centre, Osaka, Japan). 29.Ng, M M L, Application of TEM for Biomedical Sciences. Fourteenth Annual Conference of the Electron Microscopy Society of Thailand : 1996, 11-13 December 1996, J.B. Hotel, Hatyai Songkla, Thailand. 30.Ng, M M L, S.Y. Se Thoe and A.E. Ling, Immune response of dengue virus-infected human sera. Seminar on Tropical Diseases in Asia, 22-25 February 1997, Tropical Disease Research Centre, Airlangga University/Surabaya, Indonesia.

42 IV. THESES SUBMITTED a. PHD THESES 1.Studies on Rat Blastocystis. Chen Xue Qin, 1999 b. MSC THESES 1.Epitope Maping of Envelope protein of Dengue 2 Virus. MSC, Lok Shee Mei 2.Purification, characterization and cDNA cloning of phospholipase A 2 in Austrelaps superbus venom. - Simran Bedi Singh. 3.Molecular cloning, characterization and expression of short alpha-neurotoxin genes in Naja sputatrix. - Fatemeh Afifiyan. 4.Cloning and characterization of genes encoding cardiotoxins in Naja naja sputatrix. - Ramkumar Lachumanan. c. HONOURS DISERTATION 1.Immuno-modulating effects of spirulina in pracoh. Microbiology Dept. Honours Thesis. Victor Wu Jack Han (1999). 2.Immuno-modulating effects of spirulina in Prawn: Dosage effect and active component. Victor Wu Jack Han 1998/1999 3.A study on the mechanisms by which lactobacillus exclude pathogen from the human gastrointestinal tract. Lim Chai Yen 1998/1999 d. UNIVERSITY UNDERGRADUATE RESEARCH PROGRAM 1.Do Acantameoba have a role in transmission of pathogens. - Denise Teo

43 b) Symposia and Workshops July 1996 to March 1999 Training courses were conducted for the technologists in the Faculty as part of the EM Units continual education programme. CM120 TEM BioTwin with EDX and cryo-holder Freeze Fracture Replica and Etching Image Analysis and Processing Field Emission Gun Scanning Electron Microscopy 3-Dimension Reconstruction and Macro-programming 208S TEM JEM1220 with ISIS EDX Low Vacuum Scanning Electron Microscopy BioScan Digital Camera System Atomic Force Microscopy

44 Symposia The NUS-FEI/Philips Electron Optics Symposium – biennial event The NUS- JEOL Electron Microscopy Seminar –biennial event Commercial courses on : Cryo-Electron Microscopy Basic Scanning Electron Microscopy Field Emission Gun Scanning Electron Microscopy 208S TEM were conducted from 1998/99 onwards. Participants were not only from Singapore but also from as far as Australia and China. April 1999 to March 2000 Symposia The 5 th NUS-FEI/Philips Electron Optics symposium (planned for Nov 1999) was postpone this year as it was considered too close the 7 th APEM conference (June 2000). This event would now take place as one of the pre-conference activities on 22 June 2000. Workshops with FEI/Philips Electron Optics

45 a) Basic Scanning Electron Microscopy (11 to 15 October 1999) Dr. Eric Barthel from FEI Electron Optics (The Netherlands) was the instructor. Micky Leong the EM Units technologist was assisting in this course. The participants were: Micky Leong (Electron Microscopy Unit, NUS) Cau Kok Hing (Hewlett Packard in full Singapore Pte. Ltd.) Kow Roland (Institute Material Research Engineering) See Cheng Huat (Singapore Technology Microelectronics Pte Ltd.)

46 b) Field Emission Gun Scanning Electron Microscopy and EDX (18 to 21 October 1999) Dr. Barthel continued as the instructor together with Micky Leong as the assisting technologist. There were more participants for this workshops. They were: Ang Eng Chuan [Seagate Technology International (Singapore)] Lau Yi Ming (CICFAR – in full, Faculty of Engineering, NUS) Chu Heng Kian (HP- in full Singapore Pte Ltd..) Kong Lee Yong (Penang Seagate Industries) Ponniah David (Penang Seagate Industries) Tan Chee Ming (Penang Seagate Industries) Ang Ghim Boon (Chartered Semiconductor manufacturing Ltd..) Demonstration Hands-on Session

47 The benefits of being a trainer at EM Unit

48 c) Cryo-Electron Microscopy Course The cryo-Electron Microscopy course was postponed this year. It will be conducted as one of the workshops in the Pre-Conference Workshops of 7 th APEM Conference (22 to 24 June, 2000) d) Engineers Technical Training Course The Engineers Technical Training Course on the 208S TEM (17 to 21 January 2000) in conjunction with FEI/Philips Electron Optics. This is the first time a technical course of this nature has been conducted away from the factory in the Netherlands. Two factory engineers, Mr. Coert Kea and Dr. Petr Strelec were the instructors. The course is meant for the maintenance engineers and it involved disassembly and re-assembly of this microscope during the training course. It is an extensive course involving both theory and practical which started at 8.30 am until 9 p.m. each day. Patricia Netto from the EM Unit also participated in this course although she is not an engineer by training. However, she came in second out of her class of four in her test on the last day of the course. Good for her! The other three participants are the engineers from FEI/Philips Electron Optics. Participants of the Engineers Technical Training Course with Dr. Petr Strelec

49 The foyer area of the EM Unit was converted into a make shift classroom for this course. The students hard at work trying to understand what the instructor is explaining The students looking for the circuit diagrams while frantically following the instructions

50 The trainer hard at work trying to understand what the instructor (Prof Mary) is explaining

51 e) Demonstration by Philips Singapore and JEOL Asia During the period covered by this report, our microscopes were also used on some occasions by the above vendors for demonstrations to their potential customers. The demonstrations were conducted mostly by their own specialists although on a couple of occasions our technologists had to conduct the demonstrations. A charge is levied for the use of the instruments. Demonstrations were given to the following establishments (see end of users list 1999/2000 for names) f) Filming by Premier 12 One episode from the new Big Q scientific series for children was filmed in the EM Unit. The episode is called the Dirty fly. A scanning electron microscope was used to illustrate the hairy nature of the legs of a fly. Microorganisms and dirt were seen trapped among the hairs and this is the mode of transmission of these microorganisms and dirt onto the food. High magnification images of the bacteria and fungi were shown in this episode. The episode was aired on 26 February 2000, 0930hrs on Central.

52 Visitors

53 g) Visitors The Unit received the following visitors: Dr. Wim Busing – FEI Company, Holland A/P CH Diong – Division of Biology, School of Science, Nayang Technological University Mr. Badrol Ahmad – Tenaga National Research Sdn Bhd Mr. Edwin Tay – Tenaga National Research Sdn Bhd Mr. Jun Wakayama – General Manager Asia Dept., JEOL Ltd.., Tokyo Japan Mr. Jan Vrysen – FEI Company, Holland Mr. Nakhorm Techaphaithoan – FEI Company, Thailand Mr. Kensiro Kohri - JEOL Datum Mr. Ken Horie – JEOL Datum Mr. Nizamil Fairuz – University technology Malaysia Ms Normalawati Shamzudin – University Kebangsaan Malaysia Ms Zubaidah Abu Hassan – University Malaya Ms Azilah Abdul Jalil – Institute of Bioscience, Universitu Putra Malaysia Dr. Fauziah Othman – Biomedical Department, Faculty of Medicine, University Putra Malaysia Dr. Haji Mohd Noh Jahil – MARDI, Malaysia Dr. Hing Hiang Lian – University Kebangsaan Malaysia Dr. Mohd Hair Bejo – Faculty of Veterinary Medicine, University Putra Malaysia Mr. Ian Cotton - Gatan Inc. Ms Wang Yit Chee – Singapore Science Centre Ms Chen Meaw Eng – Singapore Science Centre Ms Chris Tye – Singapore Science Centre Dr. Cheong Kam Khow – Singapore Science Centre Dr. Stetan David – Leica, Austria A/P Jimmy Kwang – Institute of Molecular Agrobiology Professor David Mckenzie – University of Sydney, Australia Dr. Marcela Bilek – University of Cambridge, United Kingdom Ms Khim Hoe – Monash University, Australia Mr. Jia Yumin – FEI Company, Hong Kong Mr. Coert Kea – FEI Electron Optics, Netherlands Dr. Petr Strelec – FEI Electron Optics, Czech Republic Visitors from Malaysia

54 On-going Activities

55 ONGOING ACTIVITIES a) 7th Asia-Pacific Electron Microscopy (APEM) Conference, 26 to 30 June 2000 This conference is only a few months away. The Unit has been successful in attracting more than 25 companies relevant to microscopy for the technical exhibitions that will be held in conjunction with the conference. In total about 50 booth space have been booked. The anticipated number of delegates is at least 500 and this includes 45 invited speakers from many eminent Institutions world wide.

56 b) Handbook on Environmental Scanning Electron Microscopy (ESEM) in Life Sciences for the Novice. This Handbook will be the first publication project of the Unit. It will consist of the dos and donts encountered with this relatively new technology for life science specimens. It will be a pictorial presentation of images taken under different environmental conditions. A comparison will be made between conventional scanning electron microscopy images and that from the ESEM. It is hoped that this publication will be ready by mid-2000 to coincide with the 7 th APEM conference, as this would be a good venue to launch the handbook. 6 torr 4 torr 8 torr

57 c) The Unit has previously begun negotiating to conduct an "EMBO (European Molecular Biology Organisation) type" course on "Cryo-techniques for microscopy". Most of the instructors for this course will be attending the 7 th APEM and will visit our Unit to see our facility. From there we can then gauge the feasibility of this collaboration. d) Discussion is under way with the Australian Key Centre for Microscopy and Microanalysis, University of Sydney and Carl Zeiss Singapore to conduct a training course on Image Analysis. Expertise from the Key Centre and our Unit will be combined to conduct this course with sponsorship from Carl Zeiss. This will probably be a traveling workshop, initially within the ASEAN region, with the possibility of expanding into the Asia-Pacific area. A charge will be levied for this course and the profit shared among all the parties contributing to this course. Anticipated timing for this course will be November 2000. Report compiled : ML Ng


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