INTRO

As part of a post-treatment temperature survival trial with Westcott Pacific oysters (Crassostrea gigas) (Wikipedia), Ariana collected ctenidia tissue from 39 oysters for a resazurin assay. Steven asked that I collect ctenidia tissue from the same oysters for assessing glycogen content.

This is part of the sormi-assay-development (GitHub repo).

MATERIALS & METHODS

Ctenidia Sampling

Oysters were shucked by Ariana and she collected ctenidia samples for her resazurin assays from a total of 39 oysters. After she shucked 16 oysters, I collected and weighed ctenidia tissues for the glycogen assay. Tubes had been previously filled with homogenization buffer (see Homogenization section below). This was repeated until all 39 were samples. Samples were stored on ice prior to homogenization.

Sample labels were plate IDs (e.g. A1, A2, A3, etc.) which were tied to Ariana’s resazuring assay.

Note

Weights were to the nearest milligram.

Homogenization

The Glycogen-Glo Kit (Promega; PDF) will be used for quantifying glycogen in the ctenidia samples. Ctenidia tissues were processed in 1.5mL Safe-Lock snap cap tubes (Qiagen):

500uL PBS
250uL 0.3N HCl
~300uL 0.5mm glass beads

Samples were homogenized in a pre-cooled (dry ice) Bullet Blender Gold 5+, using the 1.5mL tube adapters. Samples were processed at Speed 12 for 4mins. After homogenization, add 250uL of Tris buffer to each, vortexed, and stored @ -20^oC.

Warning

The following samples seemed to have lost buffer during homogenization (which is weird). An additional 500uL of PBS was added, but probably won’t help due to the required pH needed for the luciferin component of the assay:

RESULTS

20251210-mgig-ctenidia_weights-glycogen_assay.csv (GitHub)

plate_ID	weight(mg)
A01	25
A02	50
A03	14
A04	21
A05	24
A06	25
A07	17
A08	12
B01	20
B02	31
B03	15
B04	28
B05	27
B06	35
B07	17
B08	31
C01	23
C02	16
C03	15
C04	24
C05	10
C06	19
C07	35
C08	35
D01	25
D02	22
D03	40
D04	23
D05	20
D06	28
D07	23
D08	25
E01	26
E02	9
E03	25
E04	31
E05	17
E06	7
E07	11