2.2.5 Quantification
and Characterization
of the Purified Virus
Particles by Flow
Cytometry
1. 24-well adherent cell culture plate (Nunc™ Thermo Fisher
Scientific, Waltham, MA, USA).
2. Host cells, infectable by the virus (Vero cells, ATCC, Manassas,
VA, USA).
3. Cell culture medium (DMEN, Sigma-Aldrich, Hamburg,
Germany).
4. PBS (Biochrom, Berlin, Germany).
5. Pipette tips 100, 1000 μL (Sarstedt, Nu ¨mbrecht, Germany).
6. Piston-operated pipettes 100 and 1000 μL (Eppendorf, Hamburg, Germany).
7. Sterile single-use pipette tips (serological pipettes, Sarstedt,
Nu ¨mbrecht, Germany).
8. Pipette holder/rubber bulb (Pipetus, Hirschmann Laborger€ ate, Eberstadt, Germany).
9. 50-mL centrifuge tubes (Sarstedt, Nu ¨mbrecht, Germany).
10. 2-mL reaction tubes (Sarstedt, Nu ¨mbrecht, Germany).
11. Virus standard at 1E+8 infective particles (in-house standard).
12. CO 2 -incubator (HeraCell 240, Thermo Fisher Scientific, Waltham, MA, USA).
13. Trypsin–EDTA solution (0.25%/0.02%, Merck, Darmstadt,
Germany).
14. Fetal calf serum (FCS) (Merck, Darmstadt, Germany).
15. 96-well U-bottom plate (Nunc™ Thermo Fisher Scientific,
Waltham, MA, USA).
16. Centrifuge with plate rotor (Multifuge 3L-R, Heraeus, Hanau,
Germany).
17. Flow cytometer (Guava
® easyCyte, Millipore Merck, Darmstadt, Germany).
18. Deionized water (Milli-Q Academic, Q-Gard
® 1, Merck
KGaA, Darmstadt, Germany).
19. Guava instrument cleaning fluid (ICF, Merck, Darmstadt,
Germany).
20. 0.5-mL and 1.5-mL tubes for waste, water and ICF (Sarstedt,
Nu ¨mbrecht, Germany).
21. Zetasizer Nano ZS90 (Malvern Panalytical, Malvern, United
Kingdom).
22. Semi-micro cuvettes, polystyrol (Sarstedt, Nu ¨mbrecht,
Germany).
23. PBS (Biochrom, Berlin, Germany).
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