Comparing Murray Clear and TDE

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I want to compare mounting with Murray Clear and TDE and decide what is the best option for analysing the morphology of Terebratalia. Murray seems to clear the yolk while TDE only optimizes the refraction index of the immersion liquid. For this I got Terebratalia late larvae and will stain them with phallacidin and propidium iodide.

  • Got Terebratalia late larvae (6 and 7 days).
  • 3x quick washes in 0.2% PTx.
  • 2x 5 min 0.2% PTx.
  • 2x 10 min PBT.
  • Incubated 2h using 1.25 U in 250 µL of Phallacidin (6.25 µL of stock) and 1:500 dilution of propidium iodide.
  • Mount in Murray Clear and TDE.
  • Check staining in the confocal.

Murray Clear + Phall/PI

MAX_Ttra TDE vs Murray - Phall-PI.lif - Series007 Murray-3

Murray Clear embryos shrank a lot and the morphology looks quite altered. Other than that, embryos are clearer and phallacidin works.

TDE + Phall/PI

MAX_Ttra TDE vs Murray - Phall-PI.lif - Series015 TDE-3

TDE is imcompatible with phallacidin and we get a artefact general cytoplasm staining. However, the morphology is quite preserved and you can scan the bottom regions of the sample well.