QX-102 QX-10232

QuantomiX.Ltd

QuantomiX Ltd.

12 Hamada Street, Tamar Science Park, rhovot POB4034, Nes-Ziona 70400, Israel

Tel:+972-8-9462244, Fax:+972-8-9465874

http://www.quantomix.com

QX-102 Applications Manual, Issue2.0

http://www.technosaurus.co.jp/support/qx-manual.htm

5

Chapter1 Introduction 7

7

7

QX-102 8

Chapter2 Liquid Samples 9

Chapter3 Particles in Solution 11

11

Poly-L-Lysine 11

13

Chapter4 Biological Applications 15

15

QX-102 17

18

19

Poly-L-Lysine 20

21

21

22

23

25

26

26

27

28

29

29

QX-102 QX-1024 5

a.　

b.

MSDS(Material Safety Data Sheet)

30

30

PTA 32

33

35

37

40

Appendix A Glossary 41

Appendix B Troubleshooting 43

QX-102 QX-1026 7

Chapter1 Introduction

QX-102

QX-102 QX-102

support@technosaurus.co.jp

Chapter/appendix Heading Provides

1 Introduction QX-1022 Liquid Sample QX-102

3 Particles in Solution

4 Biological Applications

A GlossaryB Troubleshooting

QX-102 QX-1028 9

QX-102

QX-102

wetSEMTM

QX EDS electron dispersive spectroscopy X

SEM EDS QX

QX-102

QX-102

Chapter2 Liquid Samples

QX-102

1

2 15 L

3

4 QX-102 Chapter3

Notes

a.

b.

c.

QX-102 QX-10210 11

Chapter3 Particles in Solution

QX-102

Chapter4

Poly-L-Lysine Poly-L-Lysine

Poly-L-LysinePoly-L-Lysine

Poly-L-Lysine

0.1% w/v Poly-L-lysine in water (for example Sigma Cat.No.P8920)

Distilled water

QX-102 QX-10212 13

Poly-L-Lysine

1 0.1 Poly-L-Lysine 15 L 60min

2 2

3

QX

SEM

QX

Poly-L-Lysine

1

2 15 L 60min

96 500g 5min

3 QX 15 L

4

5 QX-102 Chapter3

Poly(sodium-4-styrenesulfonate) (PSS)

PSS

PSS

30% w/v Poly(sodium-4-styrenesulfonate) in water (for example Aldrich, cat. No. 527483

Distilled water

PSS

1 30% 0.3%

2 15μl 60min

3

4

QX-102 QX-10214 15

Chapter4 Biological Applications

QX-102

QX-102

1

QX-102 QX-10216 17

QX-102

QX-102

QX-102

Poly-L-Lysine

Poly-L-Lysine

15min

0 5 2h 3 4h

1h overnight

0 5 1h

QX-102 QX-10218 19

0.1% Fibronectin solution(for example, Sigma F-1141)

PBS

1 0.01% PBS

2 0.01% 15 L 30min

3 PBS 2

4 2

5

Note

1

Gelatin(for example, Sigma Cat. No. G6144)

Distilled water

PBS

1 37 0.1% w/v

2

UV

3 0.1% 15 L 60min

4 PBS 2

5 PBS

　　　　

Notes

a

b

QX-102 QX-10220 21

Poly-L-Lysine

Poly-L-Lysine

Poly-L-Lysine

0.1% w/v Poly-L-lysine in water (for example sigman Cat. No. P8920)

Distilled water

Poly-L-Lysine

1 0.1 Poly-L-Lysine 15 L 60min

2 2

3

QX

HeLa CHO A431 NIH3T3 QX

Poly-L-Lysine

a MP-10

b MP-10

QX-102 QX-10222 23

Normal growth medium

Trypsin or non-enzymatic detachment reagent

PBS

1

QX-102

2

3 PBS

4 15 L

Table1

Note

1000 2000 sub-confl uent

5 15 L

6 MP-10 200 L

Note

MP-10

QX-102

7 5%CO2

1 0.01% 30min

2 PBS 2

3 2

4 2000

5 MP-10 200 L

6 37

QX-102

Poly-L-LysineCells/dish Cell concentration Seeding volume

1000 6.6 104/ml 15 l1500 1 105/ml 15 l2000 1.3 105/ml 15 l

Table1:

QX-102 QX-10224 25

1 Poly-L-Lysine

QX-102

2 PBS PBS

3

E.coli 100 1

4 15 L 1

96 500g 5min

5 4% PBS 10min

　 2.5% PBS 5min

6 PBS 5

QX-102

PBS

QX-102 QX-10226 27

QX-102

25% Glutaralldehyde solution EM grade(for example, Agar.Cat.No.R1020)

PBS

1 2% PBS

2 PBS 4

3 2% 30min

4 PBS 4

5

Note

0.2%～ 1%

QX-102 QX-10228 29

Paraformaldehyde, EM grade(for example,a 16% solution, Electron Microscopy Sciences, Cat. 　

　 No. 15710)

PBS

1 4% PBS

2 PBS 4

3 4% PBS 15min

4 PBS 4

5

Note

2

QX-102

QX-102

Methanol cooled to -20

PBS

1 PBS 4

2 -20

3 4 5min

4 PBS 4

QX-102 QX-10230 31

QX-102

Uranyl acetate(5% stock, pH 3.5 with HCI, kept at 4 in the dark)

Tannic Acid(2% stock in water)

4% PFA in PBS

PBS

Distilled water

0.45 m syringe fi lters

1 1%

0.5%

0.45 m

Notes

a

b

2 PBS 4

3 4% PBS 15min

P.28

4 PBS 4

5 5min4

6 1% 5min

7 5min2

8 0.5% 30min

Note

9 5min2

10 40

QX-102 QX-10232 33

PTAPTA

DNA

PTA

Phosphotungstic acid (for example sigma Cat. No. P4006), 2% stock solution in double distilled

water, pH 1.5.

Double distilled water

2% Glutaraldehyde in PBS

Notes

1

3. PBS 3

4 3

5 2%PTA 30min

6 5

7 40

Note

2% 1% PBS 30min

HeLa

QuantomiX Web Site www.quantomix.com gallary

4% OsO4(for example, Sigma Cat. No.75632)

2% Paraformaldehyde/0.1% Glutaraldehyde in PBS

Distilled water

PBS

PTA

1. PBS 4

2. 2% /PBS 30min

QX-102 QX-10234 35

Note

1 PBS 4

2 2% 0.1% PBS 30min

3 PBS 4

4 4

5 4% 0.1%

6 0.1% 30min

Note

7 4

8 40

QX-102

A

QX-102

10nm

4

2

QX-102 QX-10236 37

0.8nm 100nm

PBS

Fixative

0.2% Triton® X-100 in PBS (for intracellular antigens)

Blocking agent (BSA, normal Serum or other)

Primary antibody

Gold particle conjugate

Distilled water

Silver staining kit (for example, AURION R-GENT SE-EM, Cat.No. 500.033)

QX-102

1

2

Note

3 PBS 4

4

5 PBS 4

a

0.2%Triton X-100/PBS 10min

b PBS 5min2

6 1%BSA 5%

PBS 30min

7 1%BSA PBS

0.5h

10 min

0.5h

1h

1h

2

QX-102 QX-10238 39

Notes

a

b 30min 60min

c 37 4

8 1%BSA PBS 4

Notes

a 0.05% 0.1% Tween20

b

9 1%BSA 5%

Protein A Protein G

Note

10 PBS 4

11 30nm

Note

AURION R-GENT SE-EM kit

12 6～ 10

13 40

QX-102 QX-10240 41

QX SEM

QX PBS

QX-102 DMSO

SEM

1 QX-102

15 L

2

Notes

4

SEM

3 SEM QX-102

AppendixA

SEMMP-10

24 QX-102

MA-4

QX-102QX-102 SEM

QX-102

QX-102 SEM

Calibration Capsule

QXQX Imaging Buffer QX-102 SEM

BSA Bovine Serum Albumin

BSE Back-scattered electrons

BSED Back-scattered electrons detector

PBS Phosphate-Buffered Saline

ECM Extracelluar Matrix

PFA Paraformaldehyde

GA Glutaraldehyde

w/v Weight to Volume

QX-102 QX-10242 43

Appendix B Troubleshooting

MP-10

MA-4

MA-

QX-102

MP-10

Chapter3,

Chapter4

QX-102 QX-102

QX-102