Biomedical Engineering Reference
In-Depth Information
Aspergillus nidulans
1. Minimal medium: 5 % (v/v) salt solution, 0.1 % (v/v) trace
elements, 2 % (w/v) glucose or 2 % (v/v) glycerol; add H
2
O
bid.
,
and adjust pH with 10 M NaOH to 6.5.
2. Salt solution: 12 % NaNO
3
; 1 % KCl, 1 % MgSO
4
× 7H
2
O, 3 %
KH
2
PO
4
; add H
2
O
bid.
.
3. Trace elements: 2.2 % ZnSO
4
× 7H
2
O; 1.1 % H
3
BO
3
; 0.5 %
MnCl
2
× 4H
2
O; 0.5 % FeSO
4
× 7H
2
O; 0.16 % CoCl
2
× 5H
2
O;
0.16 % CuSO
4
× 5H
2
O; 0.11 % (NH
4
)
6
Mo
7
O
24
× 4H
2
O; 5 %
Na
4
-EDTA; adjust pH with 10 M KOH to 6.5.
4. Glass-bottom FluoroDish (World Precision Instruments,
Berlin, Germany).
1. Wide-fi eld-fl uorescence microscope (Axio Observer.Z1, Zeiss,
Jena, Germany).
2. Metal-halide lamp or laser for excitation of eGfp (488 nm/50
or 100 mW) or Rfp/mCherry (561 nm/50 or 150 mW).
3. Filter sets (eGfp ET 470/40×, ET525/50 m, T495_PXR and
mCherry ET560/40×, ET 630/75 m, T585lp, AHF
Analysentechnik, Tübingen, Germany).
4. Zeiss objectives: Plan-Apochromat (63×, NA 1.4) and EC
Plan-Neofl uar (100×, NA 1.3).
5. CoolSNAP HQ2 CCD camera (Photometrics, Tucson, AZ,
USA).
6. Software package MetaMorph (version 7, Molecular Devices,
Sunnyvale, CA, USA).
7. VisiFRAP 2D control software for Meta Series 7.6 (Visitron
Systems, Munich, Germany) to adjust laser excitation power.
8. Microscope slides 76 × 26 mm (Marienfeld, Lauda-
Königshofen, Germany).
9. High precision cover slips 18 × 18 mm,
D
= 170 ± 5
2.2 General
Microscopy Equipment
for Live Cell Imaging
ʼ
m (Zeiss,
Jena, Germany).
10. Agarose (Bio-Rad, Munich, Germany).
1. A VS-LMS4 Laser-Merge-System (Visitron Systems, Munich,
Germany) with solid-state lasers for excitation of eGfp
(488 nm/50 or 100 mW) and Rfp/mCherry (561 nm/50 or
150 mW).
2. Multiband fi lter set for eGfp/mCherry (59022, Chroma,
Bellows Falls, VT, USA).
3. VisiFRAP 2D-System (Visitron Systems, Munich, Germany)
including a 405 nm/80 mW diode laser, regulated by a UGA-40
controller (Rapp OptoElectronic GmbH, Hamburg, Germany).
4. VisiFRAP 2D control software for Meta Series 7.6 (Visitron
Systems, Munich, Germany).
2.3 Microscopy
Equipment for Local
Photobleaching
and Dual-Color
Acquisition
