Histology Staging

Staging clam reproductive state using histology slides involves first taking a single cross-section through the center of the clam starting at the hinge and bisecting the animal.

On the slide, gonad material, when present, is located towards the top on either side of the stomach lumen

Staging involves identifying maturing and mature gametes based on literature descriptions and images, such as the following from Moura et al. 2017

Staging plan

Images for all clam sections were captured by Delaney Lawson. Multiple images were taken for most slides. To reduce bias in grading, letters were assigned to each replicate image at 4X magnification for each slide, and image numbers were randomly sorted. All images for each species and sex group were then reviewed twice. Where different stage numbers were assigned between reviewing rounds or where different images from the same individual where assigned different numbers, slides were reviewed manually for final stage number determination.