11. Crushed ice for incubation.
12. 1.5 mL centrifuge tubes (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
13. Sterile single-use pipette tips (e.g., serological pipettes, Sarstedt, Nu ¨mbrecht, Germany).
14. Pipette holder/rubber bulb (e.g., pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
15. Incubator (e.g., INCU-Line
® IL 23, VWR, Radnor, Pennsylvania, USA) with orbital shaker (e.g., Multitron Standard,
Infors HT, Basel, Switzerland).
16. Centrifuge (e.g., Sigma 1-16K, Sigma-Aldrich, Hamburg,
Germany).
17. Vortexer
(e.g.,
Lab
dancer,
VWR,
Radnor,
Pennsylvania, USA).
2.4 Cell Bank
This part of the chapter describes the materials used for the operational steps described in Subheading 3.4.
2.4.1 Materials
for Thawing Insect Cells
1. Spodoptera frugiperda cell suspension (1 mL) with a cell density
of 2 Â 10
7 cells/mL (e.g., TriEx Sf-9, Merck, Darmstadt,
Germany).
2. 100 mL baffled shake flask (e.g., Glasger€ atebau Ochs, Bovenden, Germany).
3. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
4. Sterile single-use pipette tips (e.g., serological pipettes, Sarstedt, Nu ¨mbrecht, Germany).
5. Pipette holder/rubber bulb (e.g., pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
6. Centrifuge (e.g., Sigma 1-16K, Sigma-Aldrich, Hamburg,
Germany).
7. Aspiration system (e.g., Vacusafe, Integra, Biebertal,
Germany).
8. 15-mL centrifuge tubes (e.g., Sarstedt, Nu ¨mbrecht,
Germany).
9. Incubator (e.g., B-line, Binder, Tuttlingen, Germany) with
orbital shaker (e.g., Celltron, Infors HT, Basel, Switzerland).
2.4.2 Materials for Cell
Counting by Trypan Blue
Staining
1. 0.4 % trypan blue (e.g., Sigma-Aldrich, Hamburg, Germany).
2. Phosphate-buffered saline (PBS, e.g., Biochrom, Berlin,
Germany).
3. Incident light microscope (e.g., DM1i, Leica, Wetzlar,
Germany).
100
Julie Harnischfeger et al.