P. generosa Male Histology Results

Goal:

Summarize the results of the ImageJ analysis of male geoduck - adults were exposued to ambient v. low pH conditions over gametogenesis. To assess reprodutive development, we measured the volume of the total acini, lumen, spermatozoa, and spermatocytes of 4-5 acini per individual.

How to measure acini-specific chacteristics?

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Follow the steps below:

(1) Draw around the acini using the freehand selection tool
	Cntrl+M to measure
  Cntrl+T to record ROI

(2) Press Edit 🡪 Clear Outside to isolate your selection

(3) Image🡪 Adjust 🡪 color Threshold to open the color threshold window

(4) Change Hue, Saturation, and Brightness to measure the following:
    Lumen – the pink area inside the acini
    Spermatozoa - dark blue sperm cells found close to the lumen
    Spermatocytes and spermatozoa – all blue area including the sperm cells near the lumen and the circular spermatocytes (lighter blue shade) that cover the majority of the acini area
      After each…
        Cntrl+M to measure
        Cntrl+T to record ROI

(5) Add the area values for total area, lumen, spermatozoa, and spermatozoa+spermatocytes to a cumulative spreadsheet.

Example jpegs:

(i) Lumen

(ii) Spermatozoa

(iii) Spermatozoa & Spermatocytes

Results

Table: Two-Way ANOVAs (treatment×time)

table displays the mean area (µm^3) of (i) total acini (ii) lumen (iii) spermatozoa (iv) spermatocytes

Figures

figure shows the the mean area (µm^3) of (i) total acini (ii) lumen (iii) spermatozoa (iv) spermatocytes