Does droplet generation result in complete cell lysis during encapsulation? What biosafety considerations should be taken into account when operating the instrument?

Answer:
No. Cell lysis during droplet generation is not complete for all encapsulated cells.

• The system uses a lysis strategy based on non-ionic surfactants within the droplet environment. While many cells are lysed after encapsulation, a fraction of droplets may still contain viable or partially intact cells, particularly for cell types that are more resistant to lysis.

• For example, certain cell types with robust membranes (including some pathogenic or structurally resilient cells) may not be fully lysed immediately upon encapsulation. As a result, viable cells may transiently exist within the water-in-oil droplets.

• However, subsequent downstream reactions, including reverse transcription and thermal steps, will further disrupt cellular integrity.
• This behavior should be considered when evaluating biosafety risks, particularly when working with infectious or hazardous samples.
• During droplet generation, a portion of encapsulated droplets may contain viable cells due to incomplete lysis at the encapsulation stage.
• Because the instrument is not a fully enclosed biosafety system, there is a potential risk associated with handling emulsions that may contain viable cells. This is particularly relevant when working with infectious samples.
• Appropriate biosafety precautions should be implemented, such as operating within a biosafety cabinet when required and following institutional biosafety guidelines.