Chick protocols

Chick staging table
Turn eggs so they lie on sides
Incubate at 37C 40-45h to HH11 (3 primary brain vesicles, heart bent to right)

Window eggs
Clean eggs with 70% EtOH
Poke hole on blunt end of egg and small hole on top of egg
Suck out 5ml of albumin from blunt end of egg with syringe to lower embryo from shell
Tape up hole on blunt end
Tape top of egg. Cut a circular window through tape and shell
Cover hole tightly with tape

Pull glass needles
Solenoid: 2
Heating: 15
Follow instructions. Clamp top and bottom such that centre of capillary tube is at solenoid
Close cover and press START
Solenoid heats up and clamp pull and break capillary tube
Store needles in Petri dish

Injection into neural tube
Load as little as 3ul into glass needle (1ug/ul DNA + 1/10 1% Fast Green or 1mM morpholino + a bit of 1ug/ul plasmid DNA)
Break off end of glass needle
Fit needle into PicoSpritzer
Inject into neural tube till entire neural tube is filled
Add filming media + 10x AA to cover embryo (2-4ml)
Place electrodes on either side of embryo
Electroporate (See bubbles), make sure electrodes don't stick to embryo
Seal up window with tape and incubate at 37C

Injection settings
35ms
6 spritzes

Electroporation settings
25V
50ms
3 pulses

Dissection
Check embryo is alive (heart is beating)
Cut around it and pick up with scoop
Transfer embryo to Hanks' BSS, Ca2+, Mg2+ free, w/o Phenol Red
Dissect out embryo by removing extra embryonic membranes