ISOLATION AND PURIFICATION OF GIARDIA-TROPHOZOITES FROM RAT INTESTINE
ISOLATION AND PURIFICATION OF GIARDIA-TROPHOZOITES FROM RAT INTESTINE
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DOI：
10.2307/3280779
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发表时间：
1981-01-01
影响因子：
1.3
通讯作者：
ERLANDSEN, SL
中科院分区：
文献类型：
--
作者：
FEELY, DE;ERLANDSEN, SL
开发了一种基于贾第虫滋养体附着在温暖表面的能力来分离它们的方法。从感染贾第鞭毛虫的大鼠小肠取粘膜刮片，悬浮在Hanks‘s平衡盐液中。通过离心法浓缩滋养体，并允许滋养体附着在37℃孵育的聚苯乙烯培养皿表面。孵化温度显著影响滋养体的恢复。在37度时附着。C、通过低温孵育将滋养体从污染的肠道碎片中分离出来。滋养体在4度时分离。C，而肠道碎片仍有粘连。分离的滋养体在37度重贴壁分离。C.扫描电子显微镜检查显示，在37度的冷温分离和再附着后，附着的滋养体和培养皿表面的污染明显减少。C.滋养体的活力，用红素-B染料排除法测定，在分离后120分钟仍保持90%。这种分离方法便于直接从小肠中回收该原虫，用于形态和实验研究。
A method for the isolation of Giardia trophozoites based on their ability to attach to warm surfaces was developed. Mucosal scrapings were obtained from the small intestine of rats infected with Giardia and suspended in Hanks'' balanced salt solution (HBSS). Trophozoites were concentrated by centrifugation and allowed to attach to the surfaces of polystyrene petri dishes incubated at 37.degree. C. Incubation temperature significantly affected the recovery of trophozoites. After attachment at 37.degree. C, trophozoites were separated from contaminating intestinal debris by incubation at cold temperature. The trophozoites detach at 4.degree. C, whereas the intestinal debris remain adherent. Then the detached trophozoites were isolated by reattachment at 37.degree. C. Examination by scanning electron microscopy revealed a marked reduction in contamination of attached trophozoites and dish surfaces after the use of cold temperature detachment and reattachment at 37.degree. C. Viability of trophozoites, as measured by erythrosin-B dye exclusion, remained > 90% up to 120 min after isolation. This method of isolation facilitates the recovery of this protozoan directly from small intestine for morphological and experimental study.