血液涂片染色架的研制与创新

为了有效解决血液涂片制作过程中染色前后的载玻片、染色必用药品和物品侵占实验平台空间及涂片染色后染色废液对环境的污染等问题,研制了一种血液涂片染色架。该血液涂片染色架主要由支撑架、若干隔板和由下至上依次设置在支撑架上的废液槽、染色平台以及放置平台组成。使用时将涂有血膜的载玻片均匀摆放在染色平台上染色,涂片染色后将隔板移动至上面的放置平台,使得染色和晾干等步骤可以同时进行;而涂片晾干时的残余液体能够顺利流动至废液槽中,不会直接滴落。通过使用该血液涂片染色架提高了血液涂片制作的质量,节省了在制作过程中等待晾干的时间,染色废液集中收集处理利于生物安全,节约了实验平台空间。     

组织切片烘漂组合桌的研制与创新

常规石蜡切片的制作过程繁琐,所用实验仪器和工具较多,大量占用实验室平台空间且导致实验室平台杂乱无章。为了克服现有技术的不足,笔者研制了一种组织切片烘漂组合桌。该组合桌主要包括桌体、设置在桌体内的水浴组件、位于水浴组件四周的切削板及在桌体侧壁上的烘烤组件。这种烘漂组合桌可以收缩于桌体内,减少空间的占用,实现了组织切片的烘片、漂片的连续化操作,提高了实验效率。     

组织切片用漂烘应急处理仪的研制及使用

为了有效解决实验室突然断电导致组织切片漂烘中断的问题,研制了一种组织切片用漂烘应急处理仪。该漂烘应急处理仪主要由支撑架和箱体组成,箱体分为漂片箱和烤片箱,使用时将已切好的组织块小蜡片一片一片地依次放入漂片箱内进行漂片,一定时间后小蜡片在水温的作用下缓缓舒展,然后用载玻片从水中将舒展好的蜡片捞出固定于载玻片之上,装入染色架后放到烤片箱进行烤片,利用乙醇或燃气进行加热达到漂烘的目的。通过使用该组织切片用漂烘应急处理仪有效保证了在突发状况下漂片和烤片流程所需要的条件,保证了实验室临时停电时实验能够有序进行。     

数字切片结合雨课堂的《家畜组织与胚胎学》实验教学探索

在数字化背景下,着手建立《家畜组织与胚胎学》实验课程数字切片库,在此基础上制作微课,将其与网络教学平台雨课堂结合,并通过改革教学模式,着力培养具有自主学习意识和创新意识的现代化和专业化人才。     

废旧石蜡的回收处理及其应用效果

将组织切片中的废旧石蜡回收,之后进行不同的处理,并通过石蜡切片实验对比其使用效果。结果显示,回收石蜡性能优于新炼制的石蜡,回收石蜡中加入5%~9%的蜂蜡效果最佳。     

废旧石蜡的回收处理及其应用效果

将组织切片中的废旧石蜡回收,之后进行不同的处理,并通过石蜡切片实验对比其使用效果。结果显示,回收石蜡性能优于新炼制的石蜡,回收石蜡中加入5%~9%的蜂蜡效果最佳。     

鸡胚脾脏石蜡切片制作及H.E.染色方法改良的探讨

石蜡切片和H.E.染色,是组织学的最基本的技术之一,至今已使用了100多年[1].石蜡切片是目前组织标本制作的方法之一,其特点是可进行多种染色,细胞及组织层次感好,且能长期保存完好的组织形态[2].石蜡切片制作和染色过程中涉及的因素很多[3],各个环节均需要操作者根据所制作的组织不同严格筛选试验条件[4].已有大量文章进行石蜡切片方法学的研究,如组织固定不充分,会导致切片灰染[5],脱蜡不彻底导致染色不良[6-7],伊红复染时,要根据使用次数调节复染的时间[8]等,但并没有专门针对胚胎组织石蜡切片制作方法探讨的相关文章.本文选取脾脏为代表器官,目的是通过摸索制作鸡胚脾脏石蜡切片中脱水、透明、浸蜡、切片与染色的最佳条件,探讨制作优质鸡胚组织石蜡切片的方法.     

收纳式涂片染色装置组合桌的研制与创新

为了克服现有技术的不足,笔者研制了一种收纳式涂片染色装置组合桌,该组合桌主要包括架体组件、托盘和桌体三部分,架体组件的支撑柱为可收缩式的,可完全收缩于桌体内,减少实验室空间的占用;将染色平台、加热平台和置物平台置于同一架体组件上,可实现涂片、染色制片的连续化操作,提高实验效率;桌体上凹设有容置槽,容置槽内设置有废液缸,当进行冲水时,冲刷后的废液可经排水口流入废液缸内,冲洗后的涂片可直接放到加热平台上进行加热,加热平台具有烘干和沥水双重功能,进一步节约实验时间。     

绵羊卵巢组织石蜡切片制作条件优化研究

为观察绵羊正常卵巢组织的形态结构，采用苏木精-伊红（hematoxylin-eosin ，HE）染色法制作绵羊卵巢组织石蜡切片。结果显示，在切片制作过程中存在的问题主要包括染色不均、染色对比不明显、过度染色、染色过浅和切片出现斑点等。在试验过程中可通过充分分化、蓝化、充分水洗等因素的控制来提高切片的染色效果，该方法可为动物卵巢及卵泡染色切片制作提供技术指导。     

驴皮肤石蜡组织切片制作技术的探讨

皮肤组织与其他组织不同,具有一定的特殊性,其特点是纤维组织多、细胞组织少、组织硬韧。与石蜡相容性差,按常规技术难获满意切片。本实验以驴的皮肤组织为实验材料,探讨制作优质驴皮肤组织切片的技术,为后续实验驴真皮中胶原蛋白的定性和定位研究奠定基础。结果表明,用常规方法制作优质的驴皮肤组织切片时在10%甲醛固定液中固定7d,低熔点蜡锅浸蜡1h,高熔点蜡锅浸蜡2h,制作出的石蜡切片的效果最好。     

Virtual microscopy in a veterinary curriculum

Teaching faculty in the University of Tennessee College of Veterinary Medicine assist students in their professional education by providing a new way of viewing microscopic slides digitally. Faculty who teach classes in which glass slides are used participate in a program called Virtual Microscopy. Glass slides are digitized using a state-of-the-art integrated system, and a personal computer functions as the "microscope." Additionally, distribution of the interactive images is enhanced because they are available to students online. The digital slide offers equivalent quality and resolution to the original glass slide viewed on a microscope and has several additional advantages over microscopes. Students can choose to examine the entire slide at any of several objectives; they are able to access the slides (called WebSlides) from the college's server, using either Internet Explorer or a special browser developed by Bacus Laboratories, Inc.,(a) called the WebSlide browser, which lets the student simultaneously view a low-objective image and one or two high-objective images of the same slide. The student can "move the slide" by clicking and dragging the image to a new location. Easy archiving, annotation of images, and Web conferencing are additional features of the system.     

区分禽流感病毒亚型诊断基因芯片的构建

制备了可同时区分AIVH5、H7、H9血凝素亚型及N1、N2神经氨酸酶亚型的基因诊断芯片。试验以重组质粒为模板,进行PCR扩增,然后用异丙醇沉淀法进行纯化,制备探针及内参基因。将探针及内参基因用点样缓冲液稀释到0.3μg/μL后用芯片点样仪将其点制到醛基化基片上,样点中心间距450μm,样点直径220μm。将点好的基片在室温干燥24h以上,后经65℃再水合10s、80℃干燥、65mj紫外线交联照射25min后,再用封闭液封闭,95℃变性处理,成功构建了能区分禽流感血凝素H5,H7,H9亚型及神经氨酸酶N1,N2亚型的检测基因芯片。在PCR扩增中标记待检样品,对制备的检测芯片进行质量检验,结果表明,制备的区分禽流感病毒亚型基因芯片可区分AIV部分亚型。     

Malassezia otitis externa in the dog: the effect of heat-fixing otic exudate for cytological analysis

This study was conducted on 32 dogs with Malassezia otitis externa to determine the effect of heat-fixing otic exudate on cytological analysis. Malassezia infection was confirmed by cytological examination of otic exudate. Otic discharge collected with cotton swabs was then rolled onto glass slides. One slide per dog was heat-fixed prior to staining; the other slide was not heat-fixed. The number of yeast in 10 oil-immersion fields (1000 x magnification) was counted for both slides from each dog. Heat-fixing did not systematically cause either increased or decreased numbers of Malassezia on cytology of otic exudate.     

Quality of tissue specimens obtained endoscopically from the duodenum of dogs and cats

OBJECTIVE: To evaluate quality of duodenal tissue specimens obtained endoscopically from dogs and cats and submitted to 1 of 2 diagnostic laboratories for evaluation. DESIGN: Case series. SAMPLE POPULATION: Slides from 50 consecutive canine and 50 consecutive feline endoscopically obtained duodenal tissue specimens submitted to laboratory 1 and 49 consecutive canine and 46 consecutive feline specimens submitted to laboratory 2. PROCEDURE: Slides were examined independently by 3 investigators, and each tissue piece on each slide was classified as clearly inadequate, questionable, or clearly adequate on the basis of 4 criteria. An overall score was then assigned to the slide. RESULTS: Slides from laboratory 1 were more likely to be scored as clearly adequate and less likely to be scored as clearly inadequate than slides from laboratory 2. Clearly adequate slides from laboratory 1 had a higher number of clearly adequate pieces of tissue than did clearly adequate slides from laboratory 2. Slides scored as clearly adequate had a higher number of individual tissue pieces than did slides scored as clearly inadequate. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the quality of endoscopically obtained duodenal tissue specimens submitted to laboratories can vary, possibly because of differences in experience of individuals collecting biopsy specimens. Results suggest that at least 8 individual tissue pieces should be submitted when performing endoscopic biopsy of the duodenum in dogs and cats.     

Determination of the number of mast cells in lymph node, bone marrow, and buffy coat cytologic specimens from dogs.

Cytologic samples of popliteal lymph node, proximal femoral bone marrow, and the buffy coat fraction of blood were obtained from 56 dogs. The number of mast cells on 1 slide of each sample was determined by microscopic examination. Eleven of 46 slides of lymph node aspirate contained mast cells (range, 1 to 16; mean, 6.4; median, 5 mast cells/slide). Fifty-one bone marrow aspirate slides were evaluated. Two of these contained a single mast cell. None of the 53 buffy coat smear slides examined contained any mast cells. These results indicated that in clinically normal dogs, a few to several mast cells may be encountered in smears of lymph node aspirate, mast cells are rare in smears of bone marrow aspirate, and mast cells are absent from smears of buffy coat.     

Determination of K88 antigens and enterotoxins of Escherichia coli strains isolated from Polish piglets with diarrhea by the use of enzyme-linked immunosorbent assays.

We studied 103 Escherichia coli strains isolated from suckling and weaned piglets with diarrhea using different ELISA tests. K88 fimbrial antigen was determined by the slide agglutination test and the ELISA inhibition method. LT and STa enterotoxins were tested directly in the microtiter plates using monoclonal antibodies. It was found that 56.3% strains possessed K88 antigen, all of which were of the K88ac type. There was 100% correlation between the slide agglutination and ELISA tests. Of the 103 strains tested 68.9% produced LT or STa or both toxins. LT-positive strains were the most common ones in both groups of piglets. All K88-positive strains were enterotoxigenic and elaborated LT (56 strains) or LT and STa (2 strains); STb production was not determined in this study. Our ELISA tests were easy to perform, specific and can be used for determination of K88 and enterotoxins in E. coli strains isolated from piglets.     

A novel method for preparing histology slides to integrate the teaching of gross and microscopic anatomy

The authors have previously reported the development of a novel technique for sampling and preparing tissue slides for routine microscopic examination, without the use of a microtome. Termed "RAMP" (Rapid Adhesive Mediated Procedure), this simple, albeit somewhat crude, technique holds promise as a method that can be used in the field by veterinary practitioners for rapid microscopic evaluations to obtain early preliminary estimates of the nature of a mass or lesion. We incorporated the use of this method into a gross anatomy course in an attempt to gauge its utility for novices in tissue sampling and histology slide preparation. By having each group of students take a tissue sample from their cadaver, the activity simulated an actual necropsy situation in which practitioners in the field might use the technique. Because students were able to follow their specimen from sampling to microscopic examination, the activity provided a valuable integration of their learning of gross and microscopic anatomy. We conducted an evaluation of the process and the resulting slides with two successive classes of students. We conclude that the RAMP method is reasonably successful in the hands of individuals not trained in tissue preparation; was well received by the students as a valuable learning tool; and could potentially yield useful histological information for practicing veterinarians. Limitations of the method are also discussed.     

A comparison of bovine seminal quality assessments using different viewing chambers with a computer-assisted semen analyzer

Computer-assisted sperm analysis of fresh and frozen-thawed bovine sperm requires proper handling and preparation, and the type of slide used in the assessment is critical if the resultant data are to be useful quality control measurements. In the present study, 4 different slide viewing chambers, a Makler chamber, a clean slide-coverslip, or a 2- or 4-cell chamber Leja slide, were compared with assess their utility in providing reliable measurements of sperm motility variables. A Hamilton-Thorne IVOS Computer-Assisted Semen Analyzer (CASA) was the instrument used to determine sperm measurements utilizing the 4 different chambers. Fifty-eight different freeze batches of bovine semen that had been collected from 47 bulls at 7 sites that sex-sort sperm using Sexing Technologies sorting criteria were incorporated into the trial. Neither the percentage of motile sperm nor the percentage of progressively motile sperm differed for the Makler chamber vs. slide-coverslip comparisons. Similarly, total and progressively motile sperm did not differ between the 2- and 4-cell chambered Leja slides. However, total and progressive motility of sperm determined with the Makler chamber and slide-coverslip were greater (P < 0.0001) than motilities recorded by the 2- or 4-cell chambered Leja slides. Based on the results, the type of viewing chamber can affect the range of sperm motility values when CASA is used for quality control evaluations of thawed, cryopreserved sex-sorted sperm samples.     

几种主要禽疫病诊断基因芯片的制备及初步应用

进行了几种主要禽疫病诊断基因芯片制备及其初步应用研究。试验分别设计和克隆鉴定了NDV、IBV、AIV和IBDV的靶基因重组质粒。以克隆的靶基因重组质粒为模板。分别进行PCR扩增制备靶基因并纯化，以基因芯片点样仪将制备的靶基因点制在氨基化的基片上，经干燥、水合、紫外线交联和洗涤后，成功制备了NDV-IBV-AIV-IBDV诊断基因芯片。试验应用CY3荧光标记制备的探针进行芯片的检验，结果表明制备的NDV-IBV-AIV-IBDV诊断基因芯片质量好，可对NDV、IBV、AIV和IBDV进行诊断检测，具有检测灵敏性好，特异性高和芯片可重复检测的优点。试验对30个临床样品进行初步应用检测，结果表明该诊断基因芯片技术与RT—PCR检测技术检出率基本一致，并具有同步诊断检测多种疫病的优点。