1. ELECTRON MICROSCOPY UNIT 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 Engineer’s Technical Training course was initiated using the Unit’s 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 Unit’s 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 7th 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 7th 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

6. Staff

7. Staff of the EM Unit
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 Unit’s 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 Hoon CM 120 BioTwinTEM/STEM/Cryo/EDX
JEM 1220 TEM/EDX
Cryo-techniques
Miss Catherine Tang Freeze Fracture/etching
ESEM
SEM specimen preparation
From left to right : (top) Patricia, Suat Hoon, Catherine, Prof Mary,
(bottom) Cynthia, Micky.

8. Miss Patricia Netto FEG-SEM/EDX
Image analyser and processor
Digital imaging
Ms Cynthia Ng 208S TEM
1200EX TEM
Cryo-techniques
Miss Micky Leong JSM 5600 LV SEM
Freeze Fracture/etching
TEM and SEM specimen processing
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.

9. Summary

10. Since we have entered into the 21st century, it might be best to summarise the acquisition of instruments of the EM Unit during the last few years of the 20th 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
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. I. National University of Singapore, Faculty of Medicine
LIST OF USERS
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 Yong Ng Leh See
Chan Yee Gek
Research Assistant : Liu Haiping Z Emmanuel Selvanayagam
Qiang Mei

17. Students :
Anita Selvakumari Jayasurya Aye Mya Mya
Cao Qiong Gao Rong
Jin RongXian K N Srinivasan
Keiko Obaro Li JingSong
Lu Xin Rong Myint Swe
Ng Chee Hon Qi Long
R Ashok BalajiS Dinesh Kumar SelvanayagamNirthanan
T Shoba Tang Fengru
Xie Jian Ping You Yimei
Wang Dong Wang Xinsheng
Wang Xu Wu Ya Jun
Zeng XiaoXia Zhang Li

18. Biochemistry Department
Staff : Chang Chan Fong Kandiah Jeyaseelan
John Candlish
Technical Staff : Lalana Youngpritisporn
Student : Khoo Keng M Arunmozhiararsi
Chua Kim Lee
Fatemeh Aririyan
Microbiology Department
Staff : Chan Soh Ha Ng Mah Lee
Ho Bow Song Keang Peng
Kevin Tan Vincent Chow
Lee Yuan Kun Yap Eu Hian
Mulkit Singh
Technical Staff : Ho Lip Chuen Loy Boon Pheng
Josephine Howe Phoon 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 Jin Lim S M, Arthur
Research Staff : Hu Yang
Orthopaedic Surgery Department
Staff : John Chia Pho W H
P Thiagarajan V Prem Kumar
Technical Staff : Hersharan Kaur d/o Balder S Yong S C
Research Assistant : Kenjiko Hasegawa Zhu Hong
Jiang Hua Barry 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 Wee Shanmugaratnam K
Lee Hin Peng Sinniah R
Lee Yock Sun Teh Ming
Technical Staff : John Chia Lam San

22. Pharmacology Department
Staff : Benny Tan Sim Meng Hwoon
Peter Wong
Technical Staff : Tang Fengru
Research Assistant : Durairaj Ponraj
Student : Loo Li Shen Wu Xiao Hua
Physiology Department
Staff : Koh D R Shazib Pervaiz
Technical Staff : Ho Chiu Han
Research Assistant : S Mohammed Ali

23. Department of Surgery Staff : James Tan Tan Tiang Lee, Walter
Kewawan Esuwaranathan Timothy Lee
Lim Thiam Chye
Research Assistant : K M Ameer Ali Zhang Li
Student : Achuth H N Carmel 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 Peng Teo Wah Koon
Research Assistant : Atul Kumar Mittal Sun 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 Ranford Li 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 Griffith Terje Dokland
Myong Ok Kwon Yin Zhan
Sundaresan, Venkatesan William 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, Connie Ng 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 Ng Fei 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) –
3. Sinniah R and T N Khan, Ultrastructural study of renal tubular basement membrane in tubulointerstitial damage. Ultrastructural Pathology (1999, Scandinavia)
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)
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) : (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) : (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 :
10. V Zaman, J Howe, M Ng and TK Goh,Ultrastructure of the Endolimax nana cyst Parasitology Research 86: (2000)

34. 11. PY Zeng, J Hua, HC Ng and B. Ho, Unchange characteristics of Helicobacter pylori during its morphological conversion Microbios 98: , 1999
12. V Zaman, M Zak. M Manzoor, J Howe, M Ng, Postcystic development of Blastocystis hominis, Parasitology Research 85: , 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) : (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) : (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) : (Austria)
16. Zaman, V, J L C Howe and M M L Ng, Post- cystic development of Blastocystis hominis. Parasitology Research, 85 (1999) : (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) : (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 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:
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,

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 6th 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, 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 A2: Immunocytochemical localization, properties roles and involvement in neurological diseases. Neuroscientist (2000). (United States) Publication no :
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 :
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 :

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 :
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 :
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 :
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 :

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, 17th 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? 8th 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. 3rd National University of Singapore - National University Hospital Annual Scientific Meeting . 28 & 29th August 1999, Singapore.
5. R. Ashok Balaji, Gopalakrishnakone. P. " Natural toxins from Marine Conus Snail - A Review" Oral presentation, 10th 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, 7th 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 3rd NUH-Faculty of Medicine Annual Scientific Meeting 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, 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 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 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. 24th World Conference of Medical Technology Vancouver, Canada 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, 24th World Conference of Medical Techology. Vancouver, Canada 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, 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 Australia : Australian Microbiology Society, August (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 (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, 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, 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, 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, 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, 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 Australia : The Horticultural Research and Development Corporation, Australia, October (Paper presented at 3rd International Conference on Mushroom Biology and Mushroom products, 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 Australia : The Horticultural Research and Development Corporation, Australia, October (Paper presented at 3rd International Conference on Mushroom Biology and Mushroom products, 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, 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, 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, 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 A2 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. July 1996 to March 1999 b) Symposia and Workshops
Training courses were conducted for the technologists in the Faculty as part of the EM Unit’s 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 April 1999 to March 2000
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
The 5th NUS-FEI/Philips Electron Optics symposium (planned for Nov 1999) was postpone this year as it was considered too close the 7th 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 Unit’s 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 7th APEM Conference” (22 to 24 June, 2000)
d) Engineer’s Technical Training Course
The Engineer’s 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.
Participants of the Engineer’s Technical Training Course with Dr. Petr Strelec
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.

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
Success!!!

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. Visitors from Malaysia
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 don’ts” 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 7th APEM conference, as this would be a good venue to launch the handbook.
4 torr
6 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 7th 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