Mzfliii fluorescent microscope

Transgenic sticklebacks were generated by microinjection of freshly fertilized eggs
as previously described (Chan et al., 2010 (link)).
Plasmids were co-injected with Tol2 transposase mRNA as described
(Fisher et al., 2006 (link); Wada et al., 2010 (link)). Mature
Tol2 mRNA was synthesized by in vitro transcription using the
mMessage mMachine SP6 kit (Life Technologies). All enhancer assays were performed on
high-plated fish derived from Little Campbell River (British Columbia), Bodega Bay
(California), or Rabbit Slough (Alaska). Microscopic observation for GFP expression
was conducted with a MZFLIII fluorescent microscope (Leica Microsystems, Bannockburn,
IL) using GFP2 filters and a ProgResCF camera (Jenoptik AG, Jena, Germany) to
distinguish GFP expression from autofluorescence in pigmented fish. We generated
stable lines by making crosses from mosaic founder animals.

Transgenic sticklebacks were generated by microinjection of freshly fertilized eggs as previously described (Chan et al., 2010 (link)). Plasmids were co-injected with Tol2 transposase mRNA as described (Hosemann et al., 2004 (link)). Mature Tol2 mRNA was synthesized by in vitro transcription using the mMessage mMachine SP6 kit (Life Technologies). All enhancer assays were performed on pelvic-complete stickleback from Matadero Creek, California, USA (MATA). All larvae were raised under standard aquarium conditions to Swarup St 29/30 (Swarup, 1958 (link)), when pelvic bud development is initiated, for phenotyping. Larvae were anesthetized in 0.0003% w/v tricaine (Ethyl 3-aminobenzoate methanesulfonate, Sigma). Microscopic observation for GFP expression was conducted with a MZFLIII fluorescent microscope (Leica Microsystems, Bannockburn, IL) using GFP2 filters and a ProgResCF camera (Jenoptik AG, Jena, Germany) to distinguish GFP expression from autofluorescence in pigmented fish.