Protocol for Research
Protocol
Materials:
* Fertilized chicken eggs (Gallus gallus) * Tumor cell line (SK-ChA-1) * 70% ethanol * Paper towel * 45 Petri dishes * Incubator * Non-fertilized eggs * Rocket fuel * Marker * Scalpels * PBS-suspension * Fine forceps * Decapitation-scissors * Plastic rings 1mm diameter * 80 mm triangular magnetic stir bar * DCCP + DSPE-PEG 96:4 mmol empty liposomes suspension * DCCP + DSPE-PEG 96:4 mmol + Zn-Phthalocyanine (Lipid:PS - 1:0,003) * Culture flask * FALCON tubes * 25 ml plastic pipette * Microscope * Syringe
Methods:
Part 1: Incubation of eggs 1. Fertilized eggs can be stored at 13° Celsius 2. Eggs are incubated for 72 hours, lying horizontally in an incubator with a tilting tray, tilting the eggs several times a day. Humidity is kept at a minimum of 80%, incubation temperature at 37.5°C.
Note: Before starting incubation, dirt, feathers and excrement are carefully removed from the egg shells mechanically by dry wiping with common, hand paper towels.
Part 2: The ex ovo culture 1. After incubation for 72 hours, eggs are removed from the incubator 2. The top side of the eggs, where the embryo resides, is marked by marker since the embryo resists rotation to a certain extent. 3. The egg is held horizontally with the marker mark on top and cracked on the edge of an 80 mm triangular magnetic stir bar laying oriented perpendicular to the long axis of the egg. No gloves should be used for a better feeling, but hands should be disinfected with 70% EtOH. It is important that the eggshell as well as the egg membrane is opened up. Leaking of egg white is a safe sign that the egg membrane has been perforated. 4. The egg is kept close to the bottom of the Petri dish to avoid further leakage of egg white. During the initial opening procedure, it is important that the egg white on the bottom of the Petri dish is still connected to the rest
Materials:
* Fertilized chicken eggs (Gallus gallus) * Tumor cell line (SK-ChA-1) * 70% ethanol * Paper towel * 45 Petri dishes * Incubator * Non-fertilized eggs * Rocket fuel * Marker * Scalpels * PBS-suspension * Fine forceps * Decapitation-scissors * Plastic rings 1mm diameter * 80 mm triangular magnetic stir bar * DCCP + DSPE-PEG 96:4 mmol empty liposomes suspension * DCCP + DSPE-PEG 96:4 mmol + Zn-Phthalocyanine (Lipid:PS - 1:0,003) * Culture flask * FALCON tubes * 25 ml plastic pipette * Microscope * Syringe
Methods:
Part 1: Incubation of eggs 1. Fertilized eggs can be stored at 13° Celsius 2. Eggs are incubated for 72 hours, lying horizontally in an incubator with a tilting tray, tilting the eggs several times a day. Humidity is kept at a minimum of 80%, incubation temperature at 37.5°C.
Note: Before starting incubation, dirt, feathers and excrement are carefully removed from the egg shells mechanically by dry wiping with common, hand paper towels.
Part 2: The ex ovo culture 1. After incubation for 72 hours, eggs are removed from the incubator 2. The top side of the eggs, where the embryo resides, is marked by marker since the embryo resists rotation to a certain extent. 3. The egg is held horizontally with the marker mark on top and cracked on the edge of an 80 mm triangular magnetic stir bar laying oriented perpendicular to the long axis of the egg. No gloves should be used for a better feeling, but hands should be disinfected with 70% EtOH. It is important that the eggshell as well as the egg membrane is opened up. Leaking of egg white is a safe sign that the egg membrane has been perforated. 4. The egg is kept close to the bottom of the Petri dish to avoid further leakage of egg white. During the initial opening procedure, it is important that the egg white on the bottom of the Petri dish is still connected to the rest