Hydrodynamics inside wells of a 96-square deepwell microplate

For 96-square deepwell microtiter plates, culture volumes between 0.5 ml and  0.75 ml are typically used, preferably at 300 rpm on a shaker with a 50 mm amplitude (photo and video on the right).

For 0.5 ml cultures, these shaking conditions lead to oxygen transfer rates (OTR) of 38 mmol O2 l-1 h-1  (corresponding to kLa values of 200 h-1). Such an OTR - in its turn - corresponds to the high end achievable in Erlenmeyer shaker flasks (culture volume 10% of total flask volume, 300 rpm), and furthermore ensures mixing down to the bottom (no dead zones). The latter is especially relevant  for large and heavy cells (such as yeasts and CHO-cells) that tend to precipitate in dead zones. Some groups use culture volumes of 0.6 ml instead of 0.5 ml, which still results in an acceptable degree of mixing.


25 mm shaker amplitude, 300 rpm

(full mixing up to 0.25 ml culture volume)

OTR:   n.d., 12*, 7*, 3* mmol O2 l-1 h-1      
* Duetz et al. (2000). Methods for intense aeration, growth, storage, and replication of bacterial strains in microtiter plates. Applied and Environmental Microbiology 66:2641-2646


50 mm shaker amplitude, 300 rpm

(full mixing up to 0.5 ml culture volume)

OTR:   n.d., 38*, 24*, 18* mmol O2 l-1 h-1    
Preferred condition


Mixing times

The mixing times under the above conditions are less than a second, as determined by adding a drop of acid to an alkaline solution (containing a pH indicator), and following the colour change with a high-speed camera (in the video below for a volume of 0.5 ml, resulting in a mixing time of 0.8 seconds).

50 mm shaker amplitude, 225 rpm


Contact us

The best way to contact us is to use the form below. However, you may also contact Wouter Duetz directly by e-mail. This e-mail address can be found on the "about us" page.

Optionally you may provide us with additional details about your project; culture type & duration, throughput and desired culture volumes.

We are happy to assist you. We generally reply within a single business day.