Inoculation; Sampling - Sartorius Stedim Biotech BIOSTAT RM 20 basic Bedienungsanleitung

4.6.2 Inoculation

4.6.3 Sampling

40 Menu Operation
4. Disinfect the Luer septum: Wipe or spray the upper part of the Luer septum with
70 % alcohol (or similar medium).
5. Connect the syringe: connect a sterile, single-use syringe without a needle to the
Luer connection under aseptic conditions.
6. Open the tubing clamp of the Luer septum.
1. Fill the CultiBag RM with the syringe contents via the Luer septum.
Then remove the syringe from the Luer septum.
2. Disinfect the Luer septum again and re-close it using the protective cover.
3. Let the remaining liquid in the tubing run back into the bioreactor. Press the
tubing together a few times to ensure that any remaining liquid runs back into
the bioreactor. Then close the tubing clamp.
4. Now restart the rocker movement. Set the speed so that a wave motion is visible
on the surface of the liquid. Decrease the speed if too much foam builds up.
Sampling is carried out in the same way as inoculation (syringe without a needle
is also used here). However, you must make sure that the cells which are inside
the tubing are removed before taking your representative sample. For this
reason, take two samples as described in this procedure. The first one you will
throw away and the second is your representative sample.
The Luer septum is designed for 50 sterile sampling and/or inoculation
procedures. It contains a silicon septum that seals the tubing automatically
when the connection is severed. The sample is also protected completely because
no aerosols can get in from the outside. At lower rocker speeds (< 15 Rocker
movements/min), the cells can settle so that the sampling may not be
representative. Therefore, the rocker speed should be increased to 15 rocker
movements/min. approx. 5-10 minutes before the sampling. Then it can be
reduced again.
If you are working with large micro carriers or large cell aggregates, you should
not use the sampling equipment because it has a relatively small opening. You
should use the tubing with the MPC connection (outside diameter 11.1 mm).