High-proof Ethanol Fixation of Larval and Juvenile Fishes for Clearing and Double Staining

摘要

The fixation of larval and juvenile fishes in 95% ethanol can be substituted for formalin and bears several advantages for morphological and molecular studies: 1) specimens clear and double stain rapidly and brilliantly; 2) otoliths are preserved; and 3) high-quality DNA is available from the tissues. We present merits and limitations of 70% ethanol and 95% ethanol as alternative fixatives to 4% buffered formalin. In particular, we compare our results of clearing and double staining teleost larvae and juveniles from these three fixatives and those that have been frozen at -20 degrees C prior to the initial fixation. With our results, we can refute the long-standing notion that ethanol-fixed specimens disintegrate during clearing and staining.