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
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-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