Evaluation of concentration using a CASA system: the 2 key mistakes to avoid (2)

03.10.2017

Mistake no. 2: Pipetting without any prior training or defined procedure

Several factors, including the rheotaxis behavior, can also have an impact on the distribution of spermatozoa during phases of sample preparation and deposition on the slide.

There are several factors that can have an impact on spermatozoa quality and quantity in the samples:

• the sample mixing technique
• the pipetting technique (direct, reverse-mode, position of the tip in the liquid, volume, etc.)
• temperature of the fluid, consumables and instruments used
• the technique used for placing the fluid on the slides
• time management from the preparation stage to the analysis

The method and the standardization of the sample preparation process through to reading the CASA is therefore crucial. You need:

• Good quality pipettes that are inspected regularly by a third-party laboratory and used with good quality pipet tips,
• A single procedure that is followed by all the technicians in the laboratory,
• A training protocol.

A detailed training course must be given when the CASA system is first installed in the laboratory but also every year after that. IMV Technologies has a technical team that specializes in training technicians to ensure they use the CASA system as efficiently as possible.

Marleen Broekhuijse illustrated this aspect very well in her thesis dated 2012, entitled “Prediction of porcine male fertility”.

Before Training

After training

Figure 1: These two graphs show the impact of training on the repeatability of measurements using the CASA system. A comparison was made before and after the training course, based on parallel reading of motility measurements for several samples. 

Figure 2: A low repeatability resulting from a lack of training will generate overestimated or  underestimated values for the number of doses, giving rise to a risk of insufficient fertility (insufficient concentration) in one case, and economic losses for the center in the second case, because of the insufficient number of doses produced.

Training course

In this context, IMV Technologies pays special attention to organizing training sessions on how to prepare samples when installing Ivos and Ceros devices in laboratories. Our technical team can also provide assistance with setting up annual refresher courses on pipetting, which are necessary to ensure optimum use of CASA systems.

IMV Technologies also organizes special sessions throughout the year in L’Aigle, one of the themes being sample preparation.