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

ANNUAL REPORTANNUAL REPORTAPRIL 1999 TO MARCH 2000APRIL 1999 TO MARCH 2000

ELECTRON MICROSCOPY UNITELECTRON MICROSCOPY UNITFACULTY OF MEDICINEFACULTY OF MEDICINE

NATIONAL UNIVERSITY OF SINGAPORENATIONAL UNIVERSITY OF SINGAPORE

ANNUAL REPORTANNUAL REPORTAPRIL 1999 TO MARCH 2000APRIL 1999 TO MARCH 2000

ELECTRON MICROSCOPY UNITELECTRON MICROSCOPY UNITFACULTY OF MEDICINEFACULTY OF MEDICINE

NATIONAL UNIVERSITY OF SINGAPORENATIONAL UNIVERSITY OF SINGAPORE

ContentsContents

• Introduction

• Staff of the EM Unit

• Summary

• Specimen Processed

• Users

• Revenue

• Activities

• Visitors

• On-going Activities

IntroductionIntroduction

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 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.

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 LeeHead, Electron Microscopy UnitFaculty of Medicinee-mail: [email protected]

StaffStaff

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 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

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 Netto FEG-SEM/EDXImage 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

SummarySummary

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 EDXFreeze Fracture Replica and Freeze Etching Image Analysis and Processing System with full range of Digital Capture SystemField Emission Gun (FEG) Scanning Electron Microscope with EDX208S Transmission Electron MicroscopeJEM 1220 with ISIS EDX SystemLow Vacuum Scanning Electron Microscope (LV-SEM)Environmental Scanning Electron Microscope (ESEM)Critical Point Dryer for SEM specimen processingHigh Resolution Coating System for FEG and STEM specimensTwo Ultramicrotomes with CCD attachments Magnetic Field Cancellation Devices costing S$13, 000 eachLab-Top Computer

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

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 DischargerHydrophobic Glow DischargerHigh Resolution Photomonitor for the ESEMThese purchases came to a total of SIN$ 32,815.13

Specimen ProcessedSpecimen Processed

NUMBER OF SPECIMENS PROCESSED

July 1996 to March 1999

TEM = 3261SEM = 1013Cryo- techniques = 334Freeze fracture replica = 131EDX analysis = 170Image Analysis = 503Slides/transparencies/presentation media = 2387

April 1999 to March 2000

TEM = 924SEM = 431Cryo- techniques = 80Freeze fracture replica = 20EDX analysis = 200Image Analysis = 672Slides/transparencies/presentation media =1228

UsersUsers

April 1999 to March 2000 – the list is enclosed below.

I. National University of Singapore, Faculty of Medicine

Anatomy DepartmentStaff : Bay Boon Huat

Charanjit KaurGopalakrishnakone PLeong Seng KeeLing Eng Ang

Technical Staff : Yick Tuck Yong Ng Leh SeeChan Yee Gek

Research Assistant : Liu Haiping Z Emmanuel SelvanayagamQiang Mei

LIST OF USERS

Students :Anita Selvakumari Jayasurya Aye Mya MyaCao Qiong Gao RongJin RongXian K N SrinivasanKeiko Obaro Li JingSongLu Xin Rong Myint SweNg Chee Hon Qi LongR Ashok BalajiS Dinesh Kumar SelvanayagamNirthananT Shoba Tang FengruXie Jian Ping You YimeiWang Dong Wang XinshengWang Xu Wu Ya JunZeng XiaoXia Zhang Li

Biochemistry DepartmentStaff : Chang Chan Fong Kandiah Jeyaseelan

John Candlish

Technical Staff : Lalana Youngpritisporn

Student :Khoo Keng M ArunmozhiararsiChua Kim LeeFatemeh Aririyan

Microbiology DepartmentStaff : Chan Soh Ha Ng Mah Lee

Ho Bow Song Keang PengKevin Tan Vincent ChowLee Yuan Kun Yap Eu HianMulkit Singh

Technical Staff : Ho Lip Chuen Loy Boon PhengJosephine Howe Phoon Meng Chee

Students :A M A Nasirudeen Tay Wee Lee, JonathanEugene Quek Se Thoe Su YunJoogal Sudhir Kumar ChandromohanLim Chai Yen Wu Jack Han, VictorLim Li Lian Wee Boon YuLim Hui Ying Yap Ann TeckLoke Shee MeiRaudhhah Abdul Rahim

Obstetrics & Gynaecology DepartmentStaff : Ratnam S S

Technical Staff : Jean Ho

Opthalmology DepartmentStaff : The Late Chew Sek Jin Lim S M, Arthur

Research Staff : Hu Yang

Orthopaedic Surgery DepartmentStaff : 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 HongJiang Hua Barry P Pereira

Student : Chan Wai Kam

Otolaryngology DepartmentStaff : Ruan Runsheng

Paediatrics DepartmentStaff : Lee Bee Wah Wai Cheung

Chua Kaw Yan

Pathology DepartmentStaff : Aileen Wee Shanmugaratnam K

Lee Hin Peng Sinniah RLee Yock Sun Teh Ming

Technical Staff : John Chia Lam San

Pharmacology DepartmentStaff : Benny Tan Sim Meng Hwoon

Peter Wong

Technical Staff : Tang Fengru

Research Assistant : Durairaj Ponraj

Student :Loo Li Shen Wu Xiao Hua

Physiology DepartmentStaff : Koh D R Shazib Pervaiz

Technical Staff : Ho Chiu Han

Research Assistant : S Mohammed Ali

Department of SurgeryStaff : 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

II. National University of Singapore, Non-Faculty of Medicine

Faculty of DentistryPreventive Dentistry DepartmentStaff : Hsu Chin-Ying Stephen

Research Assistant : Shi Hong

Restorative Dentistry DepartmentStaff : Thean Pik Yen, Hilary

Faculty of EngineeringChemical Engineering DepartmentStaff : Ting Yen Peng Teo Wah Koon

Research Assistant : Atul Kumar Mittal Sun Gang Student : Yang Ping

Faculty of Science

Biological Sciences DepartmentStaff : Hugh Tan Tiang Wah

Student : Goh Wee Kee Pauline Tan Wei Leng Stephanie Seah

Chemistry DepartmentStaff : J D Ranford Li Fong Yau, Sam

Physics DepartmentStaff : Frank Watt

III. Other Institutes / Industries and Hospitals

Defence Medical Research InstituteStaff : S M Moochnala

Institute of Molecular and Cell BiologyStaff : Heinz Horstmann

Xu Jing

Institute of Molecular AgrobiologyStaff : Megan Griffith Terje Dokland

Myong Ok Kwon Yin ZhanSundaresan, Venkatesan William ChiaWang Yue

National Cancer CentreStaff : Hui Kam Man

Wu XiaFang

National University Medical InstitutesConfocal Microscopy Lab & Flow Cytometry LabResearch Scientist : Hanry Yu

Research Officer : Xu Xi

Lab Technologists : Er Poh Nee, Connie Ng Bee Ling

Student : Chia Ser Mien

Singapore Eye Research InstituteStaff : Hu Yang

Wilson Wong

Singapore PolytechnicStaff : Anna Yeo

Tan Tock Seng HospitalStaff : Fong Sean Yean Low Chee Kuang

Jeffery Chew

Singapore General HospitalStaff : Robert Ng Fei Xun,Beatty

Users from Industries who are customers of Philips Singapore Pte Ltd3MAdaptec Manufacturing (Singapore) Pte LtdChartered Semiconductor Manufacturing LtdHewlett-Packard SingaporeQuantumVTM

Users from Industries who are customers of JEOL Asia Pte LtdGlaxo Wellcome

RevenueRevenue

REVENUE GENERATED

July 1996 to March 1999

Fund from usage of equipment S$ 157,600 Fund from contract work S$ 47,300Fund from workshops S$ 12,091Consumable S$ 34,900

Total income S$ 247,891

April 1999 to March 2000

Fund from usage of equipment S$ 92,676Fund from contract work S$ 13,343Fund from workshops S$ 5,400Consumable S$ 14,305

Total income S$ 125,724

Revenue Generated by Different Sources from April 1999 to March

2000

Hospitals Vendors

Non-Faculty Faculty

Research Institutes Teaching Institutes

ActivitiesActivities

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 = 76Conference Papers = 75Ph.D. Theses = 12MSc Theses = 8Honours Dissertation = 4UROP Projects = 4Advance Diploma = 1

April 1999 to March 2000

International Refereed Journals = 42Conference Papers = 30Ph.D. Theses = 4MSc Theses = 1Honours Dissertation = 3UROP Projects = 1

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)

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

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.

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 A2: 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

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

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 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.

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, 24th 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).

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.

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 3rd 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 3rd 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.

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

b) Symposia and Workshops

July 1996 to March 1999

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-holderFreeze Fracture Replica and EtchingImage Analysis and ProcessingField Emission Gun Scanning Electron Microscopy3-Dimension Reconstruction and Macro-programming208S TEMJEM1220 with ISIS EDXLow Vacuum Scanning Electron MicroscopyBioScan Digital Camera SystemAtomic Force Microscopy

Symposia

The NUS-FEI/Philips Electron Optics Symposium – biennial eventThe NUS- JEOL Electron Microscopy Seminar –biennial event

Commercial courses on :Cryo-Electron Microscopy Basic Scanning Electron MicroscopyField Emission Gun Scanning Electron Microscopy208S 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 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

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.)

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

The benefits of being a trainer at EM Unit

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.

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 Engineer’s Technical Training Course with Dr. Petr Strelec

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

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

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.

VisitorsVisitors

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 BhdMr. Edwin Tay – Tenaga National Research Sdn BhdMr. Jun Wakayama – General Manager Asia Dept., JEOL Ltd.., Tokyo JapanMr. Jan Vrysen – FEI Company, HollandMr. Nakhorm Techaphaithoan – FEI Company, ThailandMr. Kensiro Kohri - JEOL DatumMr. Ken Horie – JEOL DatumMr. Nizamil Fairuz – University technology MalaysiaMs Normalawati Shamzudin – University Kebangsaan MalaysiaMs Zubaidah Abu Hassan – University MalayaMs Azilah Abdul Jalil – Institute of Bioscience, Universitu Putra MalaysiaDr. Fauziah Othman – Biomedical Department, Faculty of Medicine, University Putra MalaysiaDr. Haji Mohd Noh Jahil – MARDI, MalaysiaDr. Hing Hiang Lian – University Kebangsaan MalaysiaDr. Mohd Hair Bejo – Faculty of Veterinary Medicine, University Putra MalaysiaMr. Ian Cotton - Gatan Inc.Ms Wang Yit Chee – Singapore Science CentreMs Chen Meaw Eng – Singapore Science CentreMs Chris Tye – Singapore Science CentreDr. Cheong Kam Khow – Singapore Science CentreDr. Stetan David – Leica, AustriaA/P Jimmy Kwang – Institute of Molecular AgrobiologyProfessor David Mckenzie – University of Sydney, AustraliaDr. Marcela Bilek – University of Cambridge, United KingdomMs Khim Hoe – Monash University, AustraliaMr. Jia Yumin – FEI Company, Hong KongMr. Coert Kea – FEI Electron Optics, NetherlandsDr. Petr Strelec – FEI Electron Optics, Czech Republic

Visitors from Malaysia

On-going ActivitiesOn-going Activities

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.

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.

6 torr

4 torr

8 torr

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