Lineage Analysis by Single Cell Injection of Fluorescent Dextran

1. A fixed-stage microscope (i.e., one that focuses by moving the objective lens rather than the specimen platform) with an epifluorescent attachment and an extra-long working distance x20 objective lens—a condenser and normal microscope stage are not required as a platform to hold the egg, since it can be attached to the fitment designed to hold the condenser. The microscope does not need its own light source. The embryo will be illuminated from the side with a fiber-optic.

2. A high-resolution micromanipulator, for example, a Huxley design. It is helpful to have one that has a fine axial drive on it, i.e., a drive that moves the electrode along the axis of the electrode.

3. An antivibration table or equivalent antivibration device.

4. An oscilloscope.

5. An amplifier suitable for intracellular DC recording and a current injection facility.

6. A fiber-optic light source.

7. A good micropipet-puller (e.g., Flaming/Brown Model P-87 from Sutter Instruments, Novarto, CA).

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