BioEyes

I have the incredible opportunity to observe researchers in action! The two scientists that I work with are Aprell Carr Sparks and Francis Raycroft. They are both zebrafish researchers at the University of Notre Dame. I recently got to hang out with Francis while he was injecting ~ 2 hour old zebrafish. This procedure is at such a small scale that he actually resembled a surgeon performing microscopic surgery-and I guess he was!! He first had to create his own injection needle in order to stick the zebrafish without causing harm, and then to insert a VERY SMALL amount of “stuff” into just the right spot. He did this by heating up a needle to the point where he could actually stretch the glass so that the “neck” became longer thus creating a longer, and skinnier needle.   The slang for the apparatus he used is a “puller”, and that is exactly what it does! Once the needle had been stretched, Francis then had to carefully break off the very end (it had melted together closing off the opening).

Once he had “made” his needle, he was ready to start injecting the embryos. He placed about 30 embryos in a row along the edge of a slide, and then placed them under a microscope. He then began sticking the needle through the “shell” of his zebrafish embryos, and into the yolkball. He had to do it just right or else he might injure the fish or not get the injection in the right place in order for the “stuff” to be carried throughout the body of the growing zebrafish. Here are some great pictures that Francis took:

I am sure that both Francis and Aprell find much of what they do as more commonplace or humdrum, but to me, it is an AMAZING world that I previously only knew about from books, and I just think it is pretty darn cool!

(more later about WHAT he was injecting and why:)

Be the first to like.

Like

Unlike

Surprisingly, room 208 in Jordan Hall is just not a storage area for NDeRC collaboration equipment. Last Thursday, Anita, Francis and I cleared out a table space in 208 to talk about BioEYES.

Anita was looking for new projects that high school students could do, particularly those who already had experienced BioEYES. The project needs to to fit in a 5 day week schedule, with limited time to teach the students and have the students observe results (~45 min class periods). Three major projects that were discussed included observing melanocytes, characterizing ENU-induced mutant larvae (from Hyde or Li Lab at ND), or crossing Glo fish. glofish.com

While the study of melanocytes has been reported as a good activity for K-12 classrooms (Fields et al 2009), the mutant characterization would be a novel idea. The students can incross adult heterozygous mutant parents to obtain homozygous mutant larvae (~25% of progeny). The heterozygous mutants were previously isolated in an ENU-induced mutagenesis screen for visual defects. The challenge in this project is that the phenotype of the homozygous mutants may not display until 3-5 days, thereby exceeding the classroom scheduled time. However, if the mutant phenotype is visible at 3 days, then the students can conduct a gross phenotype analysis during development, i.e., heart rate, eye morphology, pigmentation, and maybe even cell death (degeneration). The homozygous mutants have not been extensively studied in the ND labs, therefore the students would be identifying new information for the ND labs.

The Glo fish project is an attractive enhancer for BioEYES. Glo Fish are commercially available transgenic zebrafish that express fluorescent color. This color is visible in normal lighting conditions. Previously, these transgenic fish were generated as a tool to detect pollutants. The fluorescent color is driven by DNA promoters that are responsive to pollutants. Ideally, the fish would only turn fluorescent in water contaminated with a specific pollutant, i.e., one promoter may induce the red color for a specific pollutant whereas another promoter induces blue color in response to a different pollutant.

http://www.nus.edu.sg/research/rg12.php

But, I think the commercially available fish are not for testing pollution, because you can purchase them with the color expression already visible.

glofish.com

In BioEYES, the students will cross the Glo fish with normal zebrafish or with other Glo fish of different colors. They will then determine the number of colored progeny, etc. Anita is planning to try this soon, and I am curious about her results. One problem with this project is that the embryos/larvae may not show the color in under 5 days, or the color expression may be weak or faint. Also, I do not believe that crossing fish of different colors would yield “rainbow fish,” but would either yield fish of one color or a population of colors (e.g., 50% of the larvae hatched will be red, and the other 50% would be blue).

But the Glo Fish are attractive to the students, especially the younger ones….and they shed new light on BioEYES in the future.

Be the first to like.

Like

Unlike