Week 01 of the Spring Quarter Summary

This week I cultured tissue from freshwater snails and grew the primary cultures in a shared UWT incubator. This process was necessary to initialize the workflow for starting any form of primary culture.

Later in the week I microdissected epithelial tissue from a lab-reared specimin of Botryllus schlosseri and seeded the tissue on a culture plate. I did not practice the procedure in full as we were still waiting for the arrival of several antibiotic supplements. The culture the following day showed signs of contamination likely due to both the inexperience with microdissection of the tissue and because of the lack of antibiotic treatment of the whole organism prior to dissection.

Intake Protocol

On Friday, I initialized the intake workflow that I want to apply weekly for all wild specimen that I bring to lab with four tunicate colonies that had been in our recirculating system. This process looks like:

  1. Using a razor blade, remove tunicate from substrate and clean off the dorsal portion with the blade and a soft paint brush.

  2. Tie the tunicate to a 3’’ x 2’’ x 1.2 mm glass slide using cotton thread and write an identification number on the slide using a diamond tip pencil following the following format sample location two letter acronym, date in YYYYMMDD, two digit value to distinguish tunicates collected from same day and location, and generation in the F0-FN format.

  3. Place glass slide with tunicate on a glass rack contained in a humidity chamber ensuring that the slide is free of any water. The goal is to get the tunicate to adhere to the slide. Leave the tunicate in humidity chamber for at least 15 minutes.

  4. After the humidity chamber, place organism in 5x CoralRx solution. For every 100 mL of salt water, add 230 uL of CoralRx. Make sure to do all the tunicates you are dipping in one batch and not sequentially to avoid a build of toxins that parasites may potentially release.

  5. Rinse tunicate and slide thoroughly with at least 2000 mL of salt water from recirculating system. Dispose of water down the drain and not back into the recirculating system.

  6. House all tunicates separately in different containers to avoid competition.

  7. Feed and clean tunicates every two days.