一种快速、简便制作旋毛虫幼虫永久标本的方法

经甲醛、乙醇、冰醋酸溶液固定,固绿染色液染色制作旋毛虫肌幼虫标本,方法简单,制作快速,幼虫虫体清晰可见,整体色调柔和,易于观察,可长期保存。     

旋毛虫肌幼虫囊包染色标本制作方法的改进

目的探讨旋毛虫肌幼虫囊包染色标本的制作方法。方法采用醋酸卡红染色法制作旋毛虫肌幼虫囊包标本。结果醋酸卡红染色的标本,囊包轮廓清晰,囊内幼虫透明度及清晰度高,体态自然,立体感较强。结论醋酸卡红染色可用于旋毛虫肌幼虫囊包永久标本的制作。     

3种旋毛虫成虫永久标本染色方法的比较

目的探讨制作旋毛虫成虫染色标本的最佳方法。方法分别使用乙醇硼砂卡红、醋酸卡红、固绿3种染色法制作旋毛虫成虫标本。结果乙醇硼砂卡红染色的标本,颜色鲜艳,结构清晰,另外两种染液染色效果不佳。结论乙醇硼砂卡红染色可用于旋毛虫成虫永久标本的制作。     

一种旋毛虫肌幼虫囊包标本的单染制作方法

本研究采用醋酸明矾卡红染色液对旋毛虫肌幼虫囊包标本进行染色。为观察不同染色时间对制片效果的影响,分别采用4种染色方法:①染色20 h后,分色;②染色2.5 h,不分色;③染色20 h,不分色;④染色40 h后,分色。结果显示,染色方法 1制作的标本效果最佳。囊包、囊内幼虫和周围肌纤维色泽差别较大,易于观察。囊壁内外两层结构可清晰辨别,囊内幼虫典型。     

两种吸虫复染制片技术的探究

采用醋酸卡红溶液和孔雀绿水溶液,对固定后的华支睾吸虫和布氏姜片虫标本进行复合染色。醋酸卡红对两种吸虫标本进行单染。复染法使得标本虫体呈三色,消化系统、排泄系统和周围肌肉组织均为浅红色,生殖器官中的子宫呈现鲜绿色,虫体两侧卵黄腺为黄褐色,虫体结构清晰、着色适度、立体感强。醋酸卡红染液单染对吸虫睾丸组织的显现力更强。在吸虫的教学和科研工作中,结合使用单染法和复染法,观察效果将会更好。     

旋毛虫肌幼虫囊包不染色标本的制作

在医学寄生虫学实验教学中,制作旋毛虫肌幼虫囊包标本通常采用染色封制法,梭形囊包明显,但如果分色过程掌握不好,虫体轮廓不清。作者尝试制作旋毛虫肌幼虫囊包不染色封制标本,用于教学实践取得良好效果。介绍如下。 旋毛虫肌幼虫囊包取自感染旋毛虫幼虫的猪横纹肌。操作工具:眼科剪,眼科镊,载玻片,大瓶皿,显微镜,吸管,玻片标本盘等。试剂:甲醛,蒸馏水,乙醇,水杨酸甲酯,中性树胶。 将感染旋毛虫幼虫的肌肉剪成碎块,夹在两张载玻片之间,轻轻加压、捆扎,置于10％甲醛固定12～24 h。解开载玻片后再浸泡1～2h,充分固定。倾去甲醛,用蒸馏水洗3次,每     

长期保存姜片虫标本固定及染色方法的改进

经中性甲醛溶液固定,明矾卡红溶液染色和2%钾明矾分色的布氏姜片虫（Fasciolopsis buski）标本,虫体内部结构清晰、颜色鲜艳、可长期保存。与常规制片方法相比,染色效果好,制片步骤简化。     

人芽囊原虫标本染色方法的改进

人芽囊原虫标本经肖氏液固定后,用改良的哈氏苏木素染色。结果表明,标本内部结构清晰,中心体明显,与传统的铁苏木素染色方法相比,染色效果好,制片时间短,操作步骤简单。     

并殖吸虫的皮棘染色方法

并殖吸虫的皮棘,是区别该虫的重要依据之一。但在一般的制片染色中,很不容易看清它的形状、大小、排列等。本法中,皮棘非常清楚,介绍如下: 一、染色液的配制 1.醋酸付洋红:洋红(Carmine)1g,氯化铝0.5g,氯化钙4g,70％酒精100ml,冰醋酸5ml。将钙、铝、洋红逐一溶解于酒精中,最后加入醋酸,放置过夜,过滤后即可使用。 2.皮棘染色液:蒸馏水845ml,升汞50g,丙酮50ml,甲醛10ml,冰醋酸95ml,酸性品红(acid     

绦虫永久染色标本的制作技术

目的制作绦虫玻片染色标本用于教学和科研。方法用墨汁染色法制作绦虫妊娠节片玻片标本,卡红染色法制作绦虫成熟节片、头节和囊尾蚴玻片标本。结果妊娠节片、成熟节片、头节和囊尾蚴经染色制片后特征结构明显可见。如妊娠节片染色标本清晰可见染成墨汁颜色的子宫主干和分支状的子宫侧支,节片一侧边缘中部可见突出的生殖腔;成熟节片染色标本均染成红色,内有着色更深的雌雄生殖系统各一套。结论墨汁染色和卡红染色制作带绦虫染色标本效果好,可永久保存。     

Use of acetocarmines for staining of larvae of digenetic Trematoda in situ]

Qualities of microscopical pictures of digeneic larvae total slides stained with different acetocarmines were compared. Semichon's acidifying carmine for topographical staining, Schneider's and Belling's carmines for chromosomes as well as iron acetocarmine (modification of the last two) were used. The last stain permits to obtain the slides, that in respect of their colour and quality of microscopical picture may be compared with slides stained with Semichon's carmine. Contrast, colour and reaction of staining depend on concentration of iron acetate in the staining solution. Total slides of investigated larvae stained respectively with Schneider's and Belling's carmines are not enough readable.     

旋毛虫成囊前期幼虫抗原保护性免疫的研究

目的比较旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原和表面抗原对小鼠产生的免疫保护作用。方法分别用旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原、表面抗原免疫小鼠,同时设佐剂组和阴性对照组,间隔7d免疫1次,共3次。末次免疫后7d,每只小鼠用200条旋毛虫感染期幼虫经口进行攻击感染。感染后7d和30d分别检查各组小鼠肠道成虫数和肌幼虫数;用ELISA测血清中抗旋毛虫肌幼虫IgG抗体。结果虫体抗原、排泄分泌抗原、表面抗原和佐剂组的成虫减虫率分别为84．89％、89．73％、85．65％、2．57％;肌幼虫减虫率分别为71．71％、80．98％、73．66％、5．60％。排泄分泌抗原组、表面抗原组的成虫减虫率（P均〈0．05）及肌幼虫减虫率（P均〈0．01）均高于虫体抗原组。各免疫组小鼠血清IgG抗体滴度明显升高,虫体抗原组、排泄分泌抗原组和表面抗原组的几何平均倒数滴度分别为2798．89、3474．51、2984．83,分别为阴性对照组（459．32）的6．09、7．56、6．50倍。结论旋毛虫成囊前期幼虫虫体抗原、排泄分泌抗原和表面抗原均能诱导宿主产生较强的抗攻击感染保护力。成囊前期幼虫的排泄分泌抗原显示出更强的免疫原性。     

乙醇对旋毛虫幼虫活力及感染性的影响

目的观察不同体积分数的乙醇溶液对旋毛虫幼虫活力及感染性的影响。方法在体外模拟胃内环境条件下,将100条旋毛虫用不同体积分数的乙醇溶液处理,美蓝-伊红-硼砂（M.E.B）染液鉴定幼虫的活力。64只昆明小鼠随机分为8组（每组8只）,6组小鼠分别经口接种或喂饲500条用不同体积分数的乙醇溶液处理不同时间后的幼虫或含500条幼虫的肌肉,另2组分别为生理盐水处理幼虫与含幼虫肌肉的对照组,感染后7 d与40 d每组各剖杀4只小鼠,分别观察肠道雌虫数与肌幼虫数。结果肌幼虫用体积分数为0.20和0.25的乙醇溶液处理240 min,死亡率分别为0和1.4%;在体积分数为0.30、0.35、0.40、0.45、0.50、0.55和0.60的乙醇溶液处理组幼虫全部死亡所需时间分别为180、90、45、30、30、20和20 min;肌幼虫用体积分数为0.65的乙醇溶液处理1和6 min的死亡率分别为44.4%和100%。旋毛虫幼虫经不同体积分数乙醇溶液处理不同时间后的死亡率差异有统计学意义（P〈0.05）。趋势性分析结果表明,在体积分数为0.25～0.60的乙醇处理组,幼虫的死亡率随乙醇体积分数的增加及处理时间的延长而升高（P〈0．05）。不同体积分数的乙醇溶液处理不同时间的幼虫接种小鼠后7d和40d,未发现肠道成虫与肌幼虫。结论乙醇对旋毛虫幼虫有较强的杀伤作用,幼虫用体积分数≥0．35的乙醇溶液处理30min其感染性及生殖力完全丧失。     

蓝氏贾第鞭毛虫粪样刮片标本复合三色染色法的初建

目的 建立蓝氏贾第鞭毛虫患者粪样刮片标本的复合三色染色方法.方法 用聚乙烯醇固定液替代传统三色染色法中的肖氏固定液,并与病理切片Masson三色染色法相结合,对现场收集的粪样刮片标本进行染色观察.结果 蓝氏贾第鞭毛虫包囊的轮廓与内部结构清晰,着色较为丰富.结论 复合三色染色法可用于蓝氏贾第鞭毛虫标本的染色保存.     

口服三苯双脒不同疗程抗小鼠旋毛虫成囊期幼虫的效果观察

目的比较以不同疗程口服300mg/（kg.d）三苯双脒抗小鼠横纹肌中旋毛虫成囊期幼虫的效果。方法 40只8周龄BALB/c小鼠被随机均分为5组,每只小鼠口饲感染旋毛虫成囊期幼虫50条。感染后第29d,分别以不同疗程（连续给药2、4、6、8 d）口服三苯双脒300mg/（kg.d）治疗,对照组不治疗。记录小鼠健康状况。停药后第7d,颈椎脱臼处死小鼠。肌肉压片法观察小鼠膈肌、咬肌、胸肌、腓肠肌中成囊期幼虫存活情况,计数总虫数、活虫数和死虫数。另取40只8周龄BALB/c小鼠,随机均分为5组,分别用不同疗程治疗后的小鼠膈肌成囊期幼虫50条口饲感染,感染后第29d,肌肉压片法计数膈肌中成囊期幼虫。结果实验期间,各组小鼠健康状况良好,未见药物不良反应。随着疗程的增加,4个部位肌肉中幼虫总虫荷和活虫数均呈下降趋势,而死虫数呈上升趋势。与对照组相比,2 d及2 d以上疗程组膈肌、咬肌和腓肠肌中成囊期幼虫总虫数和存活虫数均显著减少（P〈0.05、P〈0.01）,6 d疗程组和8 d疗程组胸肌总虫数显著减少（P〈0.05、P〈0.01）。随着疗程的增加,膈肌、咬肌、胸肌和腓肠肌的幼虫死亡率呈上升趋势,其中6 d疗程组分别为96.16%、98.06、99.13%和98.56%（P〈0.01）,8 d疗程组为99.62%~100%（P〈0.01）。疗效验证性感染表明,6 d（37.5%）和8 d（12.5%）的感染率显著低于对照组（100%）和2 d（100%）疗程组（P〈0.01）。2 d及以上疗程组小鼠平均虫荷和平均减虫率均显著低于对照组（P〈0.01）。结论口服TBD 300mg/（kg.d）,连续给药6 d或8 d,无不良药物反应,可有效杀死肌肉中的成囊期幼虫,为适宜疗程。     

Trichrome staining for detection of intestinal protozoa a better screening method

Intestinal protozoal infections are common in our country because of poor hygiene and tropical conditions. The efficacy of trichrome staining to screen stool smear was compared with commonly used methods i.e. concentrated iodine mount and direct wet mount to test its better effectiveness. All Stool samples were first examined by routine methods i.e. direct wet mount and iodine staining. A portion of stool sample was also inoculated in vial containing polyvinyl alcohol (PVA) fixative. From PVA preserved samples, slides were prepared and stained by modified wheately's trichrome method. The results of both methods were compared and relative accuracy was calculated. 1054 stool specimens were examined and 259 parasites detected, of which 20.7% were protozoa and 3.7% helminthde. Trichrome staining detected 19.1% protozoa while routine methods detected 12.9% protozoa. For identification of protozoa, accuracy was 91.8% in favor trichrome staining and 61.8% by wet mount and iodine staining. Trichrome stained smear alone can be used as screening method in those geographic areas where protozoa infections are common.     

Development of slide-method to distinguish alive and dead mycobacteria by fluorescent staining--a trial for solving the biohazard problem in TB laboratories]

An acid-fast staining can detect mycobacteria in clinical specimens rapidly and specifically. It equally stains living and dead bacteria. It would be of more clinical use if the viability of mycobacteria in a sample was determined by the staining. In the present paper, the problems of FDA/EB staining, which detects live or dead bacteria, were solved by establishing a new technique, a slide-method. An air-dried smear of Mycobacterium bovis BCG (Tokyo 172) on a glass slide was covered by a filter paper fully soaked in the staining solution (500 micrograms FDA and 40 micrograms EB per ml PBS). This was kept in an incubator at 37 degrees C for 20 min. The filter paper was removed after incubation and the slide was examined using a fluorescent microscope with a blue filter. Live bacteria were stained greenish yellow taking the FDA stain in while dead bacteria were stained red with EB. This new slide technique eliminated the problems associated with FDA/EB staining. Moreover, stained smears appeared to be more stable compared with the conventional tube method. To overcome the biohazard problems in smear examination of tubercle bacilli, heating of the slides on a heat block at 100 degrees C for 20 min or passing air dried smears in a flame 5 to 30 times was tried to kill the bacteria. The heat-treated slides were stained with FDA/EB and the number of green and red bacteria were counted. Samples of the smeared bacteria were taken after heating and cultured on a solid medium to determine the presence of any colony-forming unit. It was found that no CFU was observed after heating and the morphology of the stained sample was the same to that before heating. These facts suggest that the above mentioned method is a simple, safe yet inexpensive diagnostic tool for mycobacterial clinical specimens.     

Comparison of variants of carbol-fuchsin solution in Ziehl-Neelsen for detection of acid-fast bacilli.

To evaluate Ziehl-Neelsen (ZN) staining using variants of carbol-fuchsin solution, duplicate smears from 416 samples were stained with ZN, one set with 1% basic fuchsin and the other 0.3%. Another set of duplicate smears from 398 samples were stained with ZN, one with 1% basic fuchsin and the other 0.1%. The coded smears were read and discrepancies resolved. All samples underwent mycobacterial culture. The sensitivity of ZN using 0.3% (65%) and 1% basic fuchsin (62%) was comparable, while it was reduced using 0.1% (74%) compared to 1% basic fuchsin (83%). Reducing the concentration of basic fuchsin below 0.3% in ZN staining was found to significantly reduce its sensitivity.