Pressure stabilizer for reproducible picoinjection in droplet microfluidic systems

Our novel pressure balancing structure stabilizes a picoinjector, allowing reagent injection into flowing droplets in channels with fluctuating pressure.

Research on the Preparation and Regeneration Condition of the Chaetomium cupreum Protoplast

The factors of species of the medium, spawn age, species of the enzyme, time of hydrolysis, hydrolysis temperature, osmotic pressure stabilizer and regeneration medium, which have important effects on the Preparation and Regeneration Condition of the Chaetomium cupreum Protoplast, were discussed in this paper. These results showed that the hydrolysis condition can be described as following: Chaetomium cupreum mycelium with spawn age about 48h, the concentration of the lytic enzymes about 2% and the time of enzymatic hydrolysis 2h. The regeneration condition of the protoplast is that: Chaetomium cupreum mycelium with the spawn age 6h, the concentration of the lytic enzymes 2%, the temperature of 25°C and the time of enzymatic hydrolysis 1.5h. We can get the Chaetomium cupreum protoplast, regenerating in the RA Medium with the sorbitol, which regeneration can be over 50%.

Isolation and regeneration of protoplasts fromPenicillium digitatum

The effects of some factors on the isolation of protoplasts fromPenicillium digitatumwere studied, including the appropriate material (young mycelia and generating spores), the concentrations of enzyme and osmotic pressure stabilizers, reaction time and reaction temperature. Results demonstrated that germinating spores were an ideal material resource for the isolation ofP. digitatumprotoplasts. Highest yield and quality ofP. digitatumprotoplasts were obtained by shaking germinating spores suspended in a solution of 10 mg/ml Lywallzyme™ dissolved in 0.7 M NaCl as osmotic pressure stabilizer at 80 rev/min and 30°C for 3.0–3.5 h. The regeneration rate of the isolated protoplasts was as high as 24.9% on double-layer Czapek medium containing 0.7 M NaCl. Additionally, observation of the protoplast release pattern showed that the protoplasts ofP. digitatumwere released primarily from the hyphal apex and occasionally from the subapical or original sites of germinating tubes. The protoplasts ofP. digitatumwere regenerated in a direct manner of either yeast-like cell development or mycelium formation.