Stabilized Ultrasound Contrast Agent

A method of making a lyoprotected ultrasound contrast agent (UCA), said method comprising the steps of:

• a) preparing a UCA comprising at least a first surfactant and a second surfactant;
• and
• b) adding a lyoprotectant to the UCA to prepare a lyoprotected UCA,
• wherein the lyoprotectant is a saccharide.

A method of making a particle-stabilized ultrasound contrast agent (UCA), said method comprising the steps of:

• a) mixing at least two surfactants in 50 ml of water where one of the surfactants is TPGS and heating said mixture until both surfactants are dissolved;
• b) cooling said mixture to room temperature while stirring rapidly until the dispersible waxy solid comes out of solution as fine particles;
• e) purging the mixture using a sterile filtered gas in an ice bath;
• d) sonicating the mixture at between 100-140 W for 1-5 minutes with constant purging;
• e) placing mixture in a separation funnel with 50 ml PBS to allow effective separation of the bubbles;
• f) discarding lower 25 ml of the solution and transferring the next 50-75 ml of solution remaining is placed in a second separation funnel;
• and
• g) washing the bubbles of desired size, collecting them, and optionally, freeze drying them for storage.