Search for: All records

This protocol outlines a method for quantifying the density of Symbiodiniaceae cells in Scleractinian coral samples which have been previously ground into a homogenous paste consisting of aragonite skeleton, coral host tissue and endosymbiotic Symbiodiniaceae cells. There are four parts to quantifying Symbiodiniaceae cell density from ground corals: 1) grind and sub-sample the coral and store at -80 °C until ready to extract, 2) separate Symbiodiniaceae cells from the coral skeleton, 3) image Symbiodiniaceae fluorescence using Countess™ II FL Automated Cell Counter, and 4) quantify cells using the software ImageJ. Note: a CY5 EVOS™ light cube is required for this method. For instructions on installing the CY5 EVOS™ light cube to your Countess™ II FL Cell Counter, see page 40 of the Countess™ II FL Cell Counter user manual. This method was originally developed by Rowan McLachlan in February 2020. Claire Juracka assisted Rowan McLachlan by conducting tests to compare this method to the traditional method using a hemocytometer and microscope. The traditional cell counting method has been reported in several publications by Grottoli's team (e.g., Rodrigues & Grottoli 2007). This protocol was written by Rowan McLachlan with the guidance of Dr. Andréa Grottoli at The Ohio State University. dx.doi.org/10.17504/protocols.io.bdc5i2y6