INTRO
Steven asked me to perform resazurin assays on “selected” juvenile Ruditapes philippinarum (Manila clam) exposed to 36oC acute heat stress. The goal of the experiment is to compare metabolic activity between the “selected” clams and control clams when exposed to heat stress.
This is the first day of a two day experiment. Day 0 corresponds to the first day of measurements, and Day 1 corresponds to the second day of measurements. The same clams were measured on both days. Day 1 notebook entry is available here:
- Day 1 Notebook Entry
MATERIALS & METHODS
Control clams (C) compared with selected clams (S) at 36oC.
Clams were processed in 12-well plates and submerged in 5.0mL of resazurin working solution stored at 4oC,prepared on 20260330 (notebook entry).
Plates were measured every 30mins on a Synergy HTX (Agilent) plate reader.
Plate layout was randomized; Steven is currently the only person who knows the treatment assignments.
- Seemed to be inconsistent heating, with interior wells usually ~1.5oC lower than rest of wells.
The following wells lost volume - possibly due to clam spitting?
- Plate A, C3
- Plate C, C4
- Plate B, A3
- Plate D, B2
After 3.5hrs, Steven removed the resazurin working solution from the wells and replaced it with 5.0mL of seawater. Plates were returned to the cold room and will be measured again tomorrow.
RESULTS
Data are available here:
At the time of this writing, there is no plate layout file available for this experiment, so it’s not possible to determine which wells correspond to which clams. Steven is working on creating a plate layout file, which will be added to the GitHub repository when it’s available.