. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 38 M. R. KAUFMAN ET Figure 2. 1 Uioiescem micmgrapli of l-/vni section of tixed and DAPI-stained tissues of the accessory nidaniental gland. Bacteria (bac) densely populate the tubule, which is composed of columnar epithelial cells indicated bv the arrow. Mas;nitication bar = 40 /jin. males could be selected, the gonad at the anterior end of the mantle cavity was first examined in thin sections by light microscopy; if it was judged to be an ovary, the sectioning was continued until the region of accessory nidamental gla
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 38 M. R. KAUFMAN ET Figure 2. 1 Uioiescem micmgrapli of l-/vni section of tixed and DAPI-stained tissues of the accessory nidaniental gland. Bacteria (bac) densely populate the tubule, which is composed of columnar epithelial cells indicated bv the arrow. Mas;nitication bar = 40 /jin. males could be selected, the gonad at the anterior end of the mantle cavity was first examined in thin sections by light microscopy; if it was judged to be an ovary, the sectioning was continued until the region of accessory nidamental gland was observed. The following juveniles were examined (age post-hatching followed by number of specimens in parentheses): 0 days (10): 46 days (I); 75 days (2): 87 days (1): 100 days (2): 129 days (1). Accessory nidamental glands from adult females were dissected and fixed in glutaraldehyde prior to embedding in either Spurr's resin as described above or LR White resin as described previously (Biggs and Epel, 1991). Transmission electron microscopy Freshly dissected adult AN gland tissue was cut into 2-mm sections and immediately placed into fixative con- taining 4 ml 50% gluteraldehyde, g HEPES (1 mAf), and 80 ml sterile seawater (pH ) and incubated on a rotator overnight at 4°C. Fixed glands were rinsed in ster- ile seawater and post-fixed in osmium tetroxide (1%) for 1 h at room temperature. After two seawater rinses (5 min each), glands were dehydrated in an acetone series (20% to 70%, 10 min each) and held overnight in 70% acetone at 4°C. The acetone dehydration was continued (70% to 100%) using acetone dehydrated with CUSO4. Spurr's resin was measured volumetrically and degassed under a vacuum. Glands were incubated in a 50:50 mix of ace- tone:Spurr"s for 2 h at room temperature, 25:75 mix for 2 h, and 100% SpuiT's overnight. Samples were then sus- pended in fresh Spun"s. placed into BEEM embedding capsules, and polymerized by baking at 70°C overnigh
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