1) How to solve liquid leakage?

Leakage is usually caused by poor sealing or improper assembly. Check the following:

1. Flat membrane installation: The membrane (light side as the active surface, blue side as the support layer) should tightly fit the microporous plate inside the barrel. Wet the membrane with buffer or deionized water to ensure it is flat, wrinkle-free, and free of bubbles.
2. Symmetrical chamber assembly: After installing the membrane, push both barrels into the chamber simultaneously and tighten the threaded caps evenly to avoid uneven pressure and sealing failure.
3. Proper Luer connection: Align the syringe with the Luer interface and tighten clockwise. Avoid bending or impacting the Luer capillary during operation to prevent deformation or misalignment.
4. O-ring inspection: If leakage persists, check and replace worn or damaged O-rings to ensure proper dynamic sealing during extrusion.

2) How to improve extrusion efficiency and sample quality?

To improve extrusion efficiency and sample quality, the following methods are recommended:

1. Gradient extrusion: Use membranes sequentially (800 nm → 400 nm → 200 nm) for stepwise extrusion to reduce particle size distribution and improve uniformity and stability.
2. Temperature-controlled extrusion: When used with the RT manual extruder (temperature range 0–100 °C), adjust the extrusion temperature according to sample properties to improve lipid or nanoparticle flowability and formation.
3. Sample pretreatment: For samples prone to aggregation, perform ultrasonic dispersion, constant-temperature water bath, or gentle shear pretreatment before extrusion to prevent membrane clogging and extend membrane life.