In Vivo Chemical Screening for Modulators of Hematopoiesis and Hematological Diseases
Figure 1
Chemical screening using zebrafish embryos. Step —Wild-type, reporter, or mutant zebrafish are crossed to obtain embryos. Step —Once reaching an investigator-specified developmental stage (usually between 0–5 days after fertilization), embryos are arrayed into multi-well plates either manually or by automation. Step —Compounds from a chemical library are added into the wells containing the embryos using a multichannel pipette or a pin-transfer device. Step —After reaching the developmental stage for phenotype manifestation, which is usually within hours to a couple of days after the compound treatment, embryos may be subjected to staining procedures, reporter, or functional assays to detect chemical-induced phenotypes or reversal of genetic phenotypes. The images shown here depict differential hematopoietic gene expression between the compound-treated (red circle) and vehicle-treated (black circle) embryos as detected by RNA in situ hybridization. Step —In vivo phenotypes can be detected by visual inspection or by automated imaging and recording. Thus, the whole screening procedure, once optimized, may be automated for high-throughput experimentation and finished within a few days. In addition, a wide range of phenotypes may be detected in vivo.
