. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 118 M. ENZIEN ET AL. Figure 6. A,B. Phase/fluorescence micrographs of DAPl-stained amoebae with nuclei in telophase. C- H. Phase and fluorescence; Autofluorescence with blue filter (490nm) of small bodies, amoebae, and cysts. Bar scales = lOjim. scribed above) were allowed to grow for two days. Several milliliters of distilled water were added to the suspension and 3 ml of this water were transferred to a centrifuge tube to which ml of glutaraldehyde was added to make the final concentration of the fixative 5%. After fi
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 118 M. ENZIEN ET AL. Figure 6. A,B. Phase/fluorescence micrographs of DAPl-stained amoebae with nuclei in telophase. C- H. Phase and fluorescence; Autofluorescence with blue filter (490nm) of small bodies, amoebae, and cysts. Bar scales = lOjim. scribed above) were allowed to grow for two days. Several milliliters of distilled water were added to the suspension and 3 ml of this water were transferred to a centrifuge tube to which ml of glutaraldehyde was added to make the final concentration of the fixative 5%. After fixation overnight at room temperature and pelleting in a clinical centrifuge, the samples were washed twice in M cacodylate buffer (10 min each wash). The pellet was post-fixed for 10 min in 2% osmium tetroxide, washed twice with distilled water, and then stained with uranyl acetate for 30 min. Samples were then washed with distilled water (10 min) and dehydrated through an ethanol series as follows: 70% (5 min), 80% (5 min), 90% (5 min), 95% (10 min), 100% 3X 10 min. For embedding, the pelleted sample was placed in pro- pylene oxide for 30 min, followed by 30 min in a 1:1 mixture of propylene oxide: Spurr's resin. After transfer- ring to a beam capsule, it was recentrifuged and left in Spurr's for 5 h after which the spent Spurr's was poured off and fresh Spurr's added; the resin was polymerized for 16hat70°C. Mat material from North Pond, Laguna Figueroa, Baja California, was fixed at the site according to the methods of Stolz( 1983). Sectioning and observations. The blocks were sec- tioned by Floyd Craft (Boston University) on a Porter Blum MT2B ultramicrotome, post-stained with 2% ura- nyl acetate for 15 min, lead citrate for 5 min, and then observed on a Philips model 410 transmission electron microscope at 20 Please note that these images are extracted from scanned page images that may have been digitally enhanced for readability - coloration and appearance
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