. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. CELL BIOLOGY 205. Figure 1. Microtubules in varnnis model cell types visuali:ctl b\ /•/'; i. c'I: Limulus amehocvies (</, </'): [»;</ hlnuj urk cnthrocvtes (e, e'). Sen urchin r\K"te initnlic apparatus (Lytechinus pictus; / /'; fertilization membrane removed h\ Ni/e\ filtration) and sea urchin sperm jltiijcllu I Arbacia punctulata; g, g') are also readilv labeled. (g\ inset): a DAPI-staineJ sperm nucleus. Also, tin unkiiomi tvpe "I spider i rub hemot \te e.\/nhils distinct MB labeling III. /;'). The thiomh
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. CELL BIOLOGY 205. Figure 1. Microtubules in varnnis model cell types visuali:ctl b\ /•/'; i. c'I: Limulus amehocvies (</, </'): [»;</ hlnuj urk cnthrocvtes (e, e'). Sen urchin r\K"te initnlic apparatus (Lytechinus pictus; / /'; fertilization membrane removed h\ Ni/e\ filtration) and sea urchin sperm jltiijcllu I Arbacia punctulata; g, g') are also readilv labeled. (g\ inset): a DAPI-staineJ sperm nucleus. Also, tin unkiiomi tvpe "I spider i rub hemot \te e.\/nhils distinct MB labeling III. /;'). The thiomhoeyle MB in c. r' \\'as labeled h\ on-slide pcrjiision of the pre-selected ft\ctl it'll. liars = 10 /juti. (PBS. pH ). then incubated in PBS containing mouse monoclo- nal anti-a and anti-/3 tuhulins (Sigma T9026. T-4026. 1:1 mass mixture) pre-bound with Zenon™ Alexa Fluor 488 Flh (Z-25002. 1:1 by mass; Molecular Probes). Representative results using a Zeiss standard epifluorescence microscope and Nikon 950 digital camera are shown in Figure 1. MBs were readily visible in dogfish erythrocytes (Fig. 1 a. a'), (inactivated dogrish thrombocytes (Fig. Ib. b': c. c'). unactivated Limulus amebocytes (granulocytes; Fig. Id. d'). and blood ark erythrocytes (Fig. le. e'): moreover, the preservation of blood cell morphology was the best obtained to date. Sea urchin zygote mitotic apparatus was well labeled (Fig. If, f). including such details as crossing astral rays in the equatorial periphery. Sea urchin sperm flagellar microtubules were brightly labeled (Fig. la, g'), as were MBs discovered in unknown types of spider crab hemocytes (Fig. 1 h. h'). In all cases, microtubules were observed in only 30 min or less after antibody application. Precautionary pre-antibody blocking (PBS-1% bovine serum albumin, 60 min) was performed in some cases, but even without it there was little background. No labeling was observed when fluorescent Flh was used without anti-tuhulin. How rapidly were
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
