Lmd laser micro dissection 7000 microscope

Exogenous genes were incorporated into embryo genomes at the one-cell stage using the Tol2 transposon system^{43 (link)}. DNA constructs (final concentration 25 ng/μl) were mixed with Tol2 mRNA (final concentration 100 ng/μl), and about 200 picoliters mixed solutions were micro-injected into embryos at the one-cell stage, following protocols described earlier^{44 (link)}. Alternatively, about 200 picoliters Cre-ERT2 mRNA (final concentration 50 ng/μl) was injected into the Tg(ubi:loxP-Eos-stop-loxP-kRASG12V-T2A-mTFP) embryos at the one-cell stage. After injection, embryos were incubated in E3 medium at 28 °C. Healthy, normal embryos were selected at 24 hpf and 3 dpf for further observation and analysis. Fluorescent imaging was performed on a Nikon Eclipse Ti microscope equipped with a C-HGFI Intensilight Fiber illuminator, and confocal imaging was acquired on a Leica SP5-STED microscope. Embryos and adult zebrafish were anesthetized first in tricaine (Sigma-Aldrich), and aligned in a small liquid drop on a cover slide for imaging. The fluorescence intensity of individual images was quantified with ImageJ64 after background subtraction. In situ hybridization and H&E samples were imaged on a Leica LMD (Laser Micro Dissection) 7000 Microscope.

Ten days post fertilization embryos and adult zebrafish were first anesthetized with tricaine. Selected tissues bearing tumor or normal tissues from control were dissected. The embryos and dissected tissues were fixed in 4% paraformaldehyde overnight at 4 °C prior to rinsing in 70% ethanol. Fixation and decalcification of adult fish were performed as previously reported^{45 (link)}. Fish samples were then sent to UCLA Translational Pathology Core Laboratory (TPCL) for sectioning and H&E staining. Stained slides were imaged on a Leica LMD (Laser Micro Dissection) 7000 Microscope.