RobertsLab
Assess Resazurin signal from open vs closed oysters
Noah, please run some tests today to see if you can tell whether oysters are open or closed during resazurin incubations.
Please do the following:
Mix up a small batch of test solution (250 mL):
- 97 mL seawater DI water with Instant Ocean adjusted to approx. 25 ppt
- 555 µL resazurin stock solution (in FTR 209 fridge on top shelf)
- 250 µL (0.5 mL) DMSO (in FTR 209 chemical cabinet; dimethyl sulfoxide)
No antibiotic is required for these tests.
Next, grab some oysters from the tank room (from the black or yellow plastic crates of "extra oysters") about 15-20mm in size (approx 20 oysters).
Put the oysters in the small cups we have been using for trials and fill with 10 mL each of your resazurin solution.
Put 10 oysters in the incubator at 42°C and 10 on the counter as control.
During a 2-4 hour incubation, check every 30-45 minutes to see if you can tell if oysters are open or closed in the cups. We will need to do this carefully, since oysters may close if you move the cup. You may try to use a strong light or flashlight to shine into the cups to see if you can tell if oysters are open or closed.
Fluorescence measurements are not required.
Please also take temperature measurements of the cups during your trial. There is a new temperature probe on the bench in FTR 213 near the supplies. Just turn Probe 1 to "on".
Our goal is to determine 1) are oysters open or closed during trials and 2) how we should assess if they are open or closed in the cups.
Thank you! Let me know if you have questions.
Revised resazurin solution recipe:
194 mL seawater DI water with Instant Ocean adjusted to approx. 25 ppt) 1,110 µL (1.11 mL) resazurin stock solution 500 µL (0.5 mL) DMSO
@Noah-Oz Please proceed with another trial and incorporate fluorescence measurements. Use the same format, sample size, and temperatures from the first trial (above).