Pandalus platyceros, a native shrimp from San Juan Island

Abstract

With the fascination in immunology and the marine life in San Juan Island, I set to investigate Pandalus platyceros, a native shrimp from San Juan Island. By looking at its hemocytes, and the vascular system of Pandalus platyceros through scanning electron microscopic (SEM) photography, fix and stained microscopy, and micro CT scan. For preparation for SEM photography, I extracted the hemolymph from the subject’s ventral carapace. The hemolymph was mixed with buffered glutaraldehyde and phosphate-buffered saline to fix and keep the integrity of the hemocyte. Hemolymph was centrifuged and washed with distilled water to dry off a stub inside the fume hood for six hours. The stub would then sputter in gold atoms for SEM photography. This method allowed me to differentiate and classify different types of hemocytes in the highest definition. Thus, these images allow for further studying of hemocyte’s exterior morphology. Although, imaging of live hemocytes does not give the best quality to study their morphology. By staining hemocytes with eosin, I was able to look at their differentiation under the 40x oil lens. This method gave hemocytes to preserve in their best condition, thus allowing me to capture the hemocytes’ lifetime and differentiation. Lastly, I used micro CT to draw a 3D depiction of the Pandalus platyceros vascular system. This method created the most difficulty because the phosphotungstic acid (PTA) staining agent took extremely slow to penetrate through the chitin wall. To resolve this, I dissected the carapace then sonicate for six hours to allow PTA to penetrate at its fastest rate. The vascular system was segmented digitally with 3DSclicer and labeled. This method wraps everything up with how the hemolymph circulates and distributes inside the subject. Ultimately tie up the vascular and immunology relationship of Pandalus platyceros.