Preparation of biological specimens

for Electron Microscopy

Serving Advanced Technology

for

TEM

CONTENTS

Introduction

I Procedure of sample preparation

of a biological specimen

I 1 Preparing a biological sample
for ultramicrotomy

4 1 Various staining solutions

4 2 Procedure of staining

I 2 Fixation of biological tissue

I 4 Conventional staining of ultra-thin sections

12

uranyl acetate, lead citrate

II Observation of single particles

and viruses by negative staining

2 1 Extraction of tissue

II 1 Contrast in negatively staining

13

2 2 Fixation

II 2 Staining procedures

13

2 3 Dehydration

2 4 Substitution

2 5 Embedding

2 6 Polymerization

III Preparation of supporting film

Appendix: Preparing solutions used in microtomy

I 3 Steps in sectioning with an ultramicrotome

3 1 Trimming

3 2 Sectioning

Setting a specimen

Making of ultrathin sections

Thickness of ultrathin section

Picking up sections

Grid types

f How to make glass knifes

g Diamond knife

III 1 Preparation of collodion supporting film
(wet method)

14

III 2 Preparation of formvar supporting film
(dry method)

14

Introduction
There are several methods to prepare biological specimens depending on the nature of the
specimen and the purpose of the study.
This presentation introduces some commonly used methods.
Please, be careful in handling chemicals used in these methods as some are toxic.

I Procedure of sample preparation of a biological specimen

I 1 Preparing a biological sample for ultramicrotomy
Tools and chemicals listed here are necessary for preparation.
(Procedure)
1.

Extraction of
tissue

(Instruments/Chemicals)
Tweezers

6.

Glass bottles

Sectioning

Razor

Ultramicrotome (Top photo)

Grid
Diamond knife (Middle photo)
( Glass knife)

2.

Fixation

Glutaraldehyde

7.

Osmium tetroxide

Staining

Phosphate buffer

( Knife maker) (Bottom photo)

Uranyl acetate

Fixation

Lead citrate

3. Dehydration

Ethanol

8. Observation

Propylene oxide

Ultramicrotome

Embedding plate

4.

Embedding

( Disposable syringes)
( Disposable beakers)
Epoxy resin
Diamond knife

5. Polymerization

Embedding plate
(Top photo),
Embedding capsule
(Middle photo)

Embedding capsule

Oven for polymerization

(Bottom photo)

Knife maker

Oven for polymerization

I Procedure of sample preparation of a biological specimen

I 2 Fixation of biological tissue

b Washing in phosphate buffer. 10 min 2-3 times

I - 2 -1 Extraction of tissue

(Liquid is exchanged 2 or 3 times every 10minutes.)

I - 2 - 2 Fixation

c Post-fixation (Fixes lipids) 1~2 hr. (4C)

Pre-fixation (fixes proteins) 1~2 hr.

1~2% OsO4 in Phosphate buffer

2.5% glutaraldehyde in Phosphate buffer

d Washing in phosphate buffer. 10 min

Glutaraldehyde solution

A)

I - 2 - 3 Dehydration

A) The extracted tissue is dipped in the

chemical fixation solution.

50% ethanol
b 70% ethanol
c 80% ethanol
d 90% ethanol

Specimen

B)

Razor

e 95% ethanol

B)Cutting the tissue with two razor blades

as indicated will prevent unnecessary
deformation of the tissue.

f 100% ethanol *
g 100% ethanol *

10 min.
10 min.
10 min.
10 min.
10 min.
20 min.
20 min.

*Water is removed through a molecular sieve

Methods of washing & dehydration

aspiration
Next
solution

C)

C) The sample is transferred to a second

glutaraldehyde solution by keeping
the droplet with tissue held between
the two razors blades by the buffers
Tweezers
surface tension.
Glass bottle
specimen
Glutaraldehyde solution

Liquid is quickly replaced before the sample can dry.

I - 2 - 4 Substitution

Preparation of Epoxy resin

Only propylene oxide (PO) 10 min 2-3 times

[In the case of TAAB EPON 812 resin]

(Change Liquid 2 or 3 times every 20minutes.)

Put EPON 812 DDSA MNA in beaker.

b Mixed-solution of PO and resin

PO : Epoxy resin = 2:1

30 min.

PO : Epoxy resin = 1:1

1 hr.

PO : Epoxy resin = 1:2

1 hr.

*The use of disposable syringes and beakers facilitates the post washing.

b Mix it well.
c Add DMP-30 and stir well.

c Only Epoxy resin 2hr. or overnight

*Since an added volume is small, the use of syringes for tuberculin is recommended.

I - 2 - 5 Embedding (The freshly mixed resin should be used.)

The specimen is placed in a well of a silicone embedding plate.

Resin is poured over the tissue. Ensure no air bubbles are
trapped within the resin.

Epoxy resin
[In the case of TAAB EPON 812 resin]
EPON 812
b DDSA
c MNA
d DMP-30

48g
19g
33g
2g

*For a specimen with various orientations, use a silicone embedding plate.

Specimen

Resin

Silicone embedding plate

Gelatinous capsule embedding capsule Beam capsule

Resin
Specimen

I Procedure of sample preparation of a biological specimen

I - 2 - 6 Polymerization
(35C 1 day)
45C 1 day
60C 1 day

Appendix Preparing solutions used in microtomy

How to prepare 2.5% glutaraldehyde solution
1. Add 40mL of distilled water to 10mL of 25% glutaraldehyde
solution (commercially available: GA). (=>5% GA solution is
prepared.)
2. Add 50mL of 0.2M phosphate buffer.
=> 100mL of 2.5% GA-0.1M phosphate buffer (pH7.4) is prepared.

How to prepare 0.2M phosphate buffer (Sorensens phosphate buffer)

Solution A: Phosphate sodium (NaH2PO4) 27.6g/L
Oven for polymerization (1 stage type)

Solution B: Phosphate sodium (Na2HPO4) 53.6g/L

Oven for polymerization (3 stage type)

A : B = 28mL : 72mL => pH7.2

A : B = 19mL : 81mL => pH7.4

How to prepare 1% osmium solution

1. Dissolve 1g of osmium crystal in 25mL of distilled water.
=> 4% osmium solution (kept in a dark place and in a refrigerator)
2. Add 5mL of distilled water and 10mL of 0.2M phosphate buffer
to 4% osmium solution (kept in a dark place and in a refrigerator)
prepared in Step 1.
=> 20mL of 1% osmium-0.1M phosphate buffer (pH7.4) is prepared.
*Preparation must be performed in a fumehood.

I 3 Steps in sectioning with an ultramicrotome

I 3 2 Sectioning
Setting a specimen

I - 3 -1 Trimming

A sample block and a knife are set on a microtome.

Expose the tissue.

Trim the tip using a razor to form a pyramidal shape.
A cutting surface is made into a square,
rectangle or trapezoid.

1mm
0.5mm

1mm

0.5mm

Examples of trimming

Square

Square

Rectangle

Ultramicrotome
(Leica EM UC6)

Trapezoid

: Ideal for when ultra-low magnification images are

Sample block

needed.

Diamond
knife

b Rectangle : Suitable when sequential sections are cut and when the

block contains both hard and soft tissue.

c Trapezoid : Ideal for keeping track of the order of sections in a ribbon.

I Procedure of sample preparation of a biological specimen

b Making of ultrathin sections

d Picking up sections

Sectioned thin specimens float and are flattened on water in the knife boat.
A) The sections floated in the surface.

Pick up thin specimens on water using a TEM grid.

A) Press Method
Place a grid on a thin specimen with a support film
side down.

Daimond knife

Viewed from top

Place a grid on a thin specimen

B) A actual section

Sections

Knife boat
(with distilled water)

Sections

Advantage : Simple and easy

Disadvantage : Thin specimen may be wrinkled
Thin specimens may be overlaid

B) Pull up Method
Place a TEM grid under thin specimens with a support film on the top.
Bring thin specimens on the grid by using an eyelash probe.

c Thickness of ultrathin section

The thickness of thin specimens may vary depending on the hardness of
resin and room temperature.

Immerse a grid in water.

It is necessary to judge the thickness by color of interference.

The interference color and thickness of an ultrathin section
The optimal thickness

Thickness

Gray

< 60 nm

Silver

60 ~ 90 nm

Gold

90 ~ 150 nm

Purple

150 ~ 190 nm

Blue

190 ~ 240 nm

Green

240 ~ 280 nm

Yellow

280 ~ 320 nm

Eyelash probe
(eyelash attached to probe head)

The optimal
thickness for
observation in a
120 kV TEM.

Advantage : No wrinkles on specimens

You can place specimens

as you desire
Disadvantage : Skill needed

e Grid types

f How to make glass knifes

Grids of various shapes are available; circular holes, square holes, hexagonal

1) Place a glass bar (6 to 10 mm (T), 25 mm (W), 400 mm (L)) on the knife

holes, slit holes, single hole and marked holes. Grids with large square or

maker and equally divide this rod into two portions.

hexagoinal holes, or with a single slot are often used to pick up ultrathin sections.
The grid sizes range from 50 to 2000 mesh, which indicates the number of holes

2) Repeat equal division to make glass squares 25 mm on edge.

per inch. Thus, the larger the value, the smaller the grid square is.
400mm
Grating
Hole
200mm

Circle holes

Square holes

Marked holes

25mm

Slit holes

Single hole

Examples of grids

C
Knife maker

It is advantageous to select an appropriately shaped grid depending

on the research.
Example) Slit holes: For sequential sections

Single hole: For ultra-low magnification observation

Material: Cu, SS, Ti, Mo, Pt, Al, etc.

Cu is used in many cases, but for chemical analysis by EDS, the grid
should not contain any elements targeted for analysis.
Be careful when using Ni grids, as this material is paramagnetic.

A) Lock the glass bar.

B) Score the glass surface.
C) Fracture along the score line.

10

I Procedure of sample preparation of a biological specimen

3) Place a 25 mm square glass block in the knifemaker with the fresh cut to

Typical diamond knives

the right.
4) Score the block along a diagonal line and by applying pressure fracture
the glass along the score line.

Ultra

Used in room temperature.

For thin sectioning.

45 rotation

Cryo dry

Used in low temperature.

For thin sectioning

Two knifes from one 25 mm square block

Ultratrim

For preparation of resin block

g Diamond knife

11

I 4 Conventional staining of ultra-thin sections

b Washing
Run water (very gently!) parallel to the surface of a grid or place the grids on

I - 4 -1 Various staining solutions

droplets of distilled water.

Uranyl acetate (stains nucleic acids)

Washing bottle dispensing

distilled water

Usually, 2~5% (w/v) uranyl acetate in water is used.

Grid held in
tweezers

*Since it easily decomposes in light and high temperatures, it should preferably be kept in
a refrigderator.

Beaker
glass

b Lead citrate (stains protein and glycogen granules)

Method of Reynolds (There are several other methods.)

1.33 g of lead nitrate, 1.76 g of sodium citrate, and 30 mL of distilled

water are mixed.

c Lead staining 10minutes

b 8 mL of 1N NaOH is added. (Transparent liquid)

c Then, distilled water is added so that the whole quantity is 50 mL.

Drop Lead citrate liquid on Parafilm. Float a TEM grid on it.

*Lead forms crystal by reacting with CO2. Put the mixed liquid in a syringe. Place silicon

*Place NaOH grains around liquid.

plug on the tip of needle and store it in a refrigerator.

Grid held in tweezers

I - 4 - 2 Procedure of staining
Grid

Uranyl acetate staining 10minutes

Drop of lead citrate

Place a droplet of uranyl acetate solution on Parafilm. Float the grid on the
droplet with the tissue down.

Parafilm

Tweezers which
sandwiched the grid
Drop of uranyl acetate

Grid

Laboratory

Grids are covered with a laboratory dish. dish

Parafilm
Parafilm
Laboratory dish

NaOH grains

d Washing
Run water (very gently!) parallel to the surface of a grid.

e Drying

Grids are covered with a laboratory dish.

Wick excess water by blotting with a piece of filter paper.

12

I Procedure of sample preparation of a biological specimen

II Observation of single particles and viruses by negative staining

II 1 Contrast in negatively staining

II 2 Staining procedures

Negative staining solutions (uranyl acetate, phosphotungstic

acid, ammonium molybdate, etc.) envelope the specimen
and upon drying form a glass-like, electron dense film that
increases the contrast of the specimens, i.e. the specimen
is viewed against a dark background.

a Apply the specimen to a TEM grid

b Mount specimen on a TEM grid

Clamp a grid with support film in a tweezer. With a pipette, apply the
specimen. Remove excess liquid with a filter paper.

Bacteriophage (Negative staining)

Electrons

c Negatively staining

Staining liquid

Apply a droplet of uranyl acetate liquid on the grid before the specimen
dries. Remove excess liquid with a filter paper.

Specimen

d Dry the specimen

13

III Preparation of supporting film

III 1 Preparation of collodion supporting film (wet method)

III 2 Preparation of formvar supporting film (dry method)

Clean a glass slide with lens paper.

Prepare distilled water.

Put distilled water (temperature 30 to 40C)
in a Bchner funnel (diameter: 10 to 15 cm).

b Immerse the glass slide in 2% (w/v) formvar solution.

b Arrange grids.
Place a wire mesh in the funnel and arrange grids
on the mesh.

c In a smooth motion, gently pull the glass slide up.

Wire stage
Arrange grids on the wire mesh.

The quicker you pull the glassslide up, the thicker the film will be.

c Using a gtlass pipet, apply a drop 0.5-2% (w/v) of collodion

It is possible to pull up
the slide glass with a
constant speed.

solution (in iso-amyl acetate) on the water solution.

After applying, the droplet quickly expands on the water and forms a thin
collodoin film after the iso-amylacetate evaporates.

d Pull out distilled water.

Slowly drain the distilled water at the bottom of the
Bchner funnel, and allow the thin film to settle
on the grids.

Drain water here.

Sandwich the slide glass.

e Dry the thin film.

Place the wire mesh with grids and the collodoin support film on a filter

Supporting film preparation device

paper and allow to air dry.

14

I Procedure of sample preparation

III Preparation
of a biological
of supporting
specimen
film

d Score the edge of the collodoin film.

g Lift the supporting film.

To help floating off the collodoin film on water, score the edges of the glass

Place a piece of Parafilm on the collodoin with the grids, and then slowly lift

slide with a razor.

it up.

Grids arranged on the floated

collodoin support film.

e Float the collodoin film.

Slowly immerse the glass slide in water with the slide at a 20 to the water
surface.

Floated collodoin support film

Place the Parafilm.

f Arrange the grids.

Arrange the grids on the film floating on the water.

Floated collodoin support film

Lift the Parafilm with the

collodoin film and grids

h Allow the grids with the collodoin support film to air dry.

15

http://www.jeol.com/
1-2 Musashino 3-chome Akishima Tokyo 196-8558 Japan Sales Division Telephone:+81-42-528-3381 Facsimile:+81-528-3386
ARGENTINA
COASIN S.A.C.I.yF.
Virrey del Pino 4071,
1430 Buenos Aires
Argentina
Telephone: 54-11-4552-3185
Facsimile: 54-11-4555-3321
AUSTRALIA & NEW ZEALAND
JEOL(AUSTRALASIA) Pty.Ltd.
Suite 1, L2 18 Aquatic Drive
- Frenchs Forest NSW 2086
Australia
Telephone: 61-2-9451-3855
Facsimile: 61-2-9451-3822
AUSTRIA
LABCO GmbH
Dr.-Tritremmel-Gasse 8,
A-3013 Pressbaum, Austria
Telephone: 43-2233-53838
Facsimile: 43-2233-53176

BANGLADESH
A.Q. CHOWDHURY SCIENCE & SYNERGY PVT. LTD.
House No. 12, Road No. 5A
Sector No. 11, Uttara Dhaka - 1230
Bangladesh
Telephone: 880-2-9980790, 8953450, 8953501
Facsimile: 880-2-8854428
BELGIUM
JEOL (EUROPE) B.V.
Planet II, Gebouw B
Leuvensesteenweg 542,
B-1930 Zaventem
Belgium
Telephone:32-2-720-0560
Facsimile:32-2-720-6134
BRAZIL
JEOL Brasil Instrumetos Cientificos Ltda.
Av. Itaberaba, 3563
02739-000 Sao Paulo, SP Brazil
Telephone: 55-11-3983 8144
Facsimile: 55-11-3983 8140
CANADA
JEOL CANADA, INC.
(Represented by Soquelec, Ltd.)
5757 Cavendish Boulevard, Suite 540,
Montreal, Quebec H4W 2W8, Canada
Telephone: 1-514-482-6427
Facsimile: 1-514-482-1929
CHILE
TECSIS LTDA.
Avenida Kennedy 5454 - Piso 5
Vitacura, Santiago, Chile
Telephone: 56-2-401-8520
Facsimile: 56-2-410-8541

CHINA
JEOL LTD., BEIJING OFFICE
Room B1110/11, Wantong New World Plaza
No. 2 Fuchengmenwai Street, Xicheng District,
Beijing 100037, P.R.China
Telephone: 86-10-6804-6321/6322/6323
Facsimile: 86-10-6804-6324
JEOL LTD., SHANGHAI OFFICE
Shanghai Equatorial Hotel Office Building 803,
65 Yanan Road West, Shanghai 200040, P.R. China
Telephone: 86-21-6248-4868/4487/4537/4404
Facsimile: 86-21-6248-4075
JEOL LTD., GUANG ZHOU OFFICE
N3104, World Trade Center Building
371-375, Huan Shi East-Road, Guang Zhou,
510095, P.R.China
Telephone: 86-20-8778-7848
Facsimile: 86-20-8778-4268
JEOL LTD., WUHAN OFFICE
Room 3216, World Trading Bldg.
686 Jiefang Street, Hankou, Wuhan, Hubei 430032
P.R.China
Telephone: 86-27-8544-8953
Facsimile: 86-27-8544-8695
JEOL LTD., CHENGDU OFFICE
1807A Zongfu Building,
NO. 45 Zhongfu Road, Chengdu, Sichuan, 610016
P.R. China
Telephone: 86-28-86622554
Facsimile: 86-28-86622564

No. 1201F034C Printed in Japan, Kp

CYPRUS
JEOL (EUROPE) SAS
Espace Claude Monet, 1 Allee de Giverny
78290, Croissy-sur-Seine, France
Telephone: 33-13015-3737
Facsimile: 33-13015-3747

KUWAIT
YIACO MEDICAL Co.K.S.C.C.
P.O. Box 435
13005-Safat, Kuwait
Telephone: 965-24842322/24814358
Facsimile: 965-24844954/24833612

EGYPT
JEOL SERIVCE BUREAU
3rd Fl. Nile Center Bldg., Nawal Street,
Dokki, (Cairo), Egypt
Telephone: 20-2-3335-7220
Facsimile: 20-2-3338-4186

MALAYSIA
JEOL(MALAYSIA) SDN.BHD.(359011-M)
205, Block A, Mezzanine Floor,
Kelana Business Center,
97, Jalan SS 7/2, Kelana Jaya,
47301 Petaling Jaya, Selangor, Malaysia
Telephone: 60-3-7492-7722
Facsimile: 60-3-7492-7723

FRANCE
JEOL (EUROPE) SAS
Espace Claude Monet, 1 Allee de Giverny
78290, Croissy-sur-Seine, France
Telephone: 33-13015-3737
Facsimile: 33-13015-3747
GERMANY
JEOL (GERMANY) GmbH
Oskar-Von-Miller-Strasse 1a, 85386
Eching, Germany
Telephone: 49-8165-77346
Facsimile: 49-8165-77512
GREAT BRITAIN & IRELAND
JEOL (U.K.) LTD.
JEOL House, Silver Court, Watchmead,
Welwyn Garden City, Herts AL7 1LT, U.K.
Telephone: 44-1707-377117
Facsimile: 44-1707-373254

GREECE
N. ASTERIADIS S.A.
56-58,S. Trikoupi Str. P.O. Box 26140
GR-10022, Athens, Greece
Telephone: 30-1-823-5383
Facsimile: 30-1-823-9567
HONG KONG
FARMING LTD.
Unit 1009, 10/F., MLC Millennia Plaza
663 King's Road, North Point, Hong Kong
Telephone: 852-2815-7299
Facsimile: 852-2581-4635
INDIA
BLUE STAR LTD. (HQ: Mumbai)
Analytical Instrments Department,
Sahas' 414/2 Veer Savarkar Marg
Prabhadery Mumbai 400 025, India
Telephone: 91-22-6666-4000
Facsimile: 91-22-6666-4001
BLUE STAR LTD. (Delhi)
Analytical Instruments Department,
E-44/12 Okhla Industrial Area,
Phase-II, New Delhi 110 020, India
Telephone: 91-11-4149-4000
Facsimile: 91-11-4149-4005
BLUE STAR LTD. (Calcutta)
Analytical Instruments Department,
7, Hare Street Calcutta 700 001, India
Telephone: 91-33-2213-4133
Facsimile: 91-33-2213-4102
BLUE STAR LTD. (Chennai)
Analytical Instruments Department,
No. 46, Garuda Building,
Cathedral Road, Chennai 600 086, India
Telephone: 91-44-4244-4000
Facsimile: 91-44-4244-4190

INDONESIA
PT. TEKNOLABindo Penta Perkasa
Komplek Gading Bukit Indah Blok I/11
JI. Bukit Gading Raya Kelapa Gading Permai,
Jakarta 14240, Indonesia
Telephone: 62-21-45847057/58/59
Facsimile: 62-21-45842729
ITALY
JEOL (ITALIA) S.p.A.
Centro Direzionale Green Office
Via dei Tulipani, 1
20090 Pieve Emanuele (MI) Italy
Telephone: 39-02-9041431
Facsimile: 39-02-90414343
KOREA
JEOL KOREA LTD.
Dongwoo Bldg. 7F, 458-5, Gil-Dong,
Gangdong-Gu, Seoul, 134-010, Korea
Telephone: 82-2-511-5501
Facsimile: 82-2-511-2635

MEXICO
JEOL DE MEXICO S.A. DE C.V.
Av. Amsterdam #46 DEPS. 402
Col Hipodromo, 06100, Mexico D.F.
Mexico
Telephone: 52-5-55-211-4511
Facsimile: 52-5-55-211-0720
PAKISTAN (Karachi)
ANALYTICAL MEASURING SYSTEM (PVT) LTD.(AMS LTD.)
14-C Main Sehar Commercial Avenue Lane 4,
Khayaban-e-Sehar,
D.H.A-VII, Karachi-75500, Pakinstan
Telephone: 92-21-35345581/35340747/35346057-8
Facsimile: 92-21-35345582
PANAMA
PROMED S.A.
Parque Industrial Costa del Este
Urbanizacion Costa del Este
Apartado 0816-01755, Panama, Panama
Telephone: 507-303-3100
Facsimile: 507-303-3115
PHILIPPINES
PHILAB INDUSTRIES INC.
7487 Bagtikan Street, SAV Makati,
1203 Metro, Manila Phillippines
Telephone: 63-2-896-7218
Facsimile: 63-2-897-7732
PORTUGAL
Izasa Portugal Lda.
R. do Proletariado, 1
2790-138 CARNAXIDE, Portugal
Telephone: 351-21-424-73-00
Facsimile: 351-21-418-60-20
RUSSIA
JEOL LTD. Moscow Office
Krasnoproletarskaya Street, 16,
Bld. 2, Entrance 5, 127473, Moscow,
Russian Federation
Telephone: 7-495-748-7791
Facsimile: 7-495-748-7793
SAUDI ARABIA
ABDULREHMAN ALGOSAIBI G.T.C. (Riyadh)
King Abdulaziz Avenue,
P.O. Box 215, Riyadh 11411, Saudi Arabia
Telephone: 966-1-479-3000
Facsimile: 966-1-477-1374
SCANDINAVIA
JEOL (SKANDINAVISKA) A.B.
Hammarbacken 6A, Box 716, 191 27 Sollentuna
Sweden
Telephone: 46-8-28-2800
Facsimile: 46-8-29-1647
SERVICE & INFORMATION OFFICE
JEOL NORWAY
Ole Deviks vei 28, N-0614 Oslo, Norway
Telephone: 47-2-2-64-7930
Facsimile: 47-2-2-65-0619
JEOL FINLAND
Ylakaupinkuja 2, FIN-02360 Espoo, Finland
Telephone: 358-9-8129-0350
Facsimile: 358-9-8129-0351
JEOL DENMARK
Naverland 2, DK-2600 Glostrup, Denmark
Telephone: 45-4345-3434
Facsimile: 45-4345-3433
SINGAPORE
JEOL ASIA PTE. LTD.
2 Corporation Road #01-12 Corporation Place
Singapore 618494
Telephone: 65-6565-9989
Facsimile: 65-6565-7552

SOUTH AFRICA
ADI Scientific (Pty) Ltd.
370 Angus Crescent,
Northlands Business Park, 29 Newmarket Road
Northriding, Ranburg, Republic of South Africa
Telephone: 27-11-462-1363
Facsimile: 27-11-462-1466

Preparation of biological specimens

for Electron Microscopy

for

TEM

SPAIN
IZASA. S.A.
Argoneses, 13, 28100 Alcobendas,
(Poligono Industrial), Madrid, Spain
Telephone: 34-91-663-0500
Facsimile: 34-91-663-0545
SWITZERLAND
JEOL (GERMANY) GmbH
Oskar-Von-Miller Strasse 1,
85386 Eching, Germany
Telephone: 49-8165-77346
Facsimile: 49-8165-77512
TAIWAN
JIE DONG CO., LTD.
7F, 112, Chung Hsiao East Road,
Section 1, Taipei, Taiwan 10023
Republic of China
Telephone: 886-2-2395-2978
Facsimile: 886-2-2322-4655
For Semiconductor Products:
JEOL TAIWAN SEMICONDUCTORS LTD.
11F-1, No. 346, Pei-Da Road, Hsin-Chu City 300,
Taiwan, Republic of China
Telephone: 886-3-523-8490
Facsimile: 886-3-523-8503

THAILAND
BECTHAI BANGKOK EQUIPMENT & CHEMICAL CO., Ltd.
300 Phaholyothin Rd. Phayathai, Bangkok 10400,
Thailand
Telephone: 66-2-615-2929
Facsimile: 66-2-615-2350/2351
THE NETHERLANDS
JEOL (EUROPE) B.V.
Lireweg 4, NL-2153 PH Nieuw-Vennep,
The Netherlands
Telephone: 31-252-623500
Facsimile: 31-252-623501
TURKEY
TEKSER LTD.STI.
Acibadem Cad. Erdem Sok, N 6/1
34660, Uskudar, Istanbul, Turkey
Telephone: 90-216-3274041
Facsimile: 90-216-3274046
UAE
BUSINESS COMMUNICATIONS LLC. (Abu Dhabi)
P.O. Box 2534, Abu Dhabi UAE
Telephone: 971-2-6348495
Facsimile: 971-2-6316465
BUSINESS COMMUNICATIONS LLC. (Dubai)
P.O. Box 233, Dubai, UAE
Telephone: 971-4-2220186
Facsimile: 971-4-2236193

USA
JEOL USA, INC.
11 Dearborn Road, Peabody, MA 01960, U.S.A.
Telephone: 1-978-535-5900
Facsimile: 1-978-536-2205/2206
JEOL USA, INC. WEST OFFICE
5653 Stoneridge Drive Suite #110
Pleasanton, CA 94588, U.S.A.
Telephone: 1-925-737-1740
Facsimile: 1-925-737-1749

VENEZUELA
GOMSA Service and Supply C.A.
Urbanizacion Montalban III
- Residencias Don Andres - Piso 7 - Apartomento 74
Avenida 3, entre calles 7 y 6
Montalban, Caracas, Venezuela
Telephone: 58-212-443-4342
Facsimile: 58-212-443-4342
VIETNAM
TECHNICAL MATERIALS AND RESOURCES IMPORTEXPORT JOINT STOCK COMPANY(REXCO)
Hanoi Branch,
No. 13-Lot 12 Trung Yen, Trung Hoa Street, Cau Giay Dist,
Hanoi, Vietnam
Telephone: 84-4-562-0516,17/562-0535
Facsimile: 84-4-853-2511

MGMT. SYS. MGMT. SYS.

R v A C 024 R vA C 42 5
ISO 9001 & 14001 REGISTERED FIRM
DNV Certification B.V., THE NETHERLANDS

ISO 9001 & ISO 14001 Certificated

Certain products in this brochure are controlled under the

Exchange and Foreign Trade Law of Japan in compliance
international security export control. JEOL Ltd. must pr
Japanese Government with End-users Statement of Assura
and End-use Certificate in order to obtain the export
needed for export from Japan. If the product to be exported
category, the end user will be asked to fill in these cert