Introduction and evaluation of virtual microscopy in teaching veterinary cytopathology

Virtual microscopy (VM) uses a computer to view digitized slides and is comparable to using a microscope to view glass slides. This technology has been assessed in human medical education for teaching histology and histopathology, but, to the authors' knowledge, no one has evaluated its use in teaching cytopathology in veterinary medical education. We hypothesize that students will respond positively to the use of VM for viewing cytopathology preparations and that the technology can be successfully used for student assessment. To test this hypothesis, we surveyed students regarding their level of satisfaction with features of the VM system, their preference for use of VM in the curriculum, and the potential influence virtual slides may have on student study habits; student performance on a traditional cytopathology practical examination and a similar exam using VM was evaluated. Our results show that student perception of the VM system is generally very positive, with some concerns about resolution and the need for continued exposure to traditional microscopy. Within the curriculum, students indicated a preference for the option of using virtual slides for studying and take-home exercises. Overwhelmingly, students wanted either hybrid laboratory sessions or sessions using glass slides with virtual slides available for study and review. Students identified many VM test-taking features as advantageous compared with traditional glass-slide practical exams as traditionally administered. However, students indicated a strong preference for continued use of traditional microscopy for graded practical exams. Students may be more likely to study slides in preparation for practical examinations if virtual slides are available. Results also indicate that VM can be used successfully for assessment purposes, but students should receive training in using virtual slides if the technology will be used for assessment.     

Teaching histology to first-year veterinary science students using virtual microscopy and traditional microscopy: a comparison of student responses

Virtual microscopy (VM) is a comparatively recent innovation that is revolutionizing both the teaching of microscopic structure in human medicine and the concept of online diagnosis and telemedicine. The interactivity of the various commercially available browsers attempts to simulate the experience of looking down a microscope while offering advantages over traditional microscopy that include clarity of image, reduced infrastructure, and high flexibility, as the images are accessible online. We developed our own VM system, including customized software and a browser that was simple and intuitive to use, with the added advantage of further modifications possible to assist student learning. In this article, we report on a preliminary study wherein VM was introduced to veterinary science students in one course and directly compared to traditional microscopy to determine whether students would readily accept this new technology and which aspects of VM were advantageous. Responses from a survey form showed that students rated VM significantly higher than traditional microscopy as a tool to learn histology because it offers clearer images, the ability to learn collaboratively, more effective use of time, and the flexibility of online learning. Students also indicated a strong preference for the use of VM in future courses. These results suggest that VM is a flexible and enjoyable resource that could be useful to enhance the learning of microscopic structure in veterinary science courses.     

Determination of Oestrous Cycle of the Rats by Direct Examination: How Reliable?

For determination of the oestrous cycle in rats classical Papanicolaou technique has long been used successfully. Instead of using many stains in Papanicolaou, staining the vaginal secretions with only methylene blue has also been defined. Recently a new technique in which vaginal samples are directly examined under light microscope has been introduced. The aim of this study was to assess the reliability of this new technique by comparing it with the classical staining techniques. From 20 Wistar rats 60 vaginal samples were collected with a micropipette, three from each. Briefly, the vagina was flushed two to three times then the fluid was placed onto a glass slide. The fluid was equally distributed onto three glass slides. The glass slides were coded. Two samples were stained with Papanicolaou and methylene blue while the other one was examined directly. Determination of the phases of the oestrous cycle was made by the same histologist who was blinded to the groups and coding system. After determination of the oestrous phase in all samples, the results were compared and it was found that the results were matching. In conclusion, the same results can be obtained with the direct examination technique and this technique is reliable, so there is no need to use relatively time-consuming, less practical and more expensive techniques such as Papanicolaou or methylene blue.     

Improvement of the teflonized slides used in the immunofluorescent antibody technique

The indirect fluorescent antibody (IF) test is often performed on glass slides prepared with smears or tissue sections containing an antigen. It is convenient to have several separate and distinct areas of antigen on the same slide, e.g. to compare dilution steps, and to save time when producing the antigen slides. With this end in view a technique for preparing slides with wells in a thin film of teflon-like compound has been elaborated (Goldman 1968). This technique is useful when a great number of slides need to be prepared in advance, e.g. to test a substantial amount of material. It has, however, not been widely used im serology for Babesia, perhaps because any teflon remaining within the wells is stained with the fluorescent dye, thus creating a potentially disturbing background.     

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.     

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。     

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.     

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.     

Motic显微数码互动实验室在遗传学实验教学中的应用

在遗传学实验教学中,Motic数码显微互动系统与实验教学相结合,以形象的画面展示教学内容,实现师生互动,使教师能够高效率地对全体学生给予指导。另外,计算机强大的图像处理等也会极大地激发学生实验兴趣,有助于实际操作能力的提高。     

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。     

组织切片烘烤架的研制及使用

为了有效解决新制作的组织切片在晾干过程中所带来的侵占实验平台空间及污染问题,研制了一种组织切片烘烤架。该组织切片烘烤架制作和使用方法简单,主要由支撑架和若干隔板组成,隔板与支撑架通过滑轨连接,使用时将新制作出来的组织切片先摆放在镂空的隔板上,再将隔板插入支撑架的滑槽内,最后放入烤箱烘烤。通过使用该组织切片烘烤架提高了切片的质量,减少了二甲苯对实验人员的伤害,节省了实验平台的空间。     

An attempt to estimate students' workload

Following the recent introduction of the European Credit Transfer System (ECTS) into several European university programs, a new interest has developed in determining students' workload. ECTS credits are numerical values describing the student workload required to complete course units; ECTS has the potential to facilitate comparison and create transparency between institutional curricula. ECTS credits are frequently listed alongside institutional credits in course outlines and module summaries. Measuring student workload has been difficult; to a large extent, estimates are based only upon anecdotal and casual information. To gather more systematic information, we asked students at the Veterinary Faculty, University of Ljubljana, to estimate the actual total workload they committed to fulfill their coursework obligations for specific subjects in the veterinary degree program by reporting their attendance at defined contact hours and their estimated time for outside study, including the time required for examinations and other activities. Students also reported the final grades they received for these subjects. The results show that certain courses require much more work than others, independent of credit unit assignment. Generally, the courses with more contact hours tend also to demand more independent work; the best predictor of both actual student workload and student success is the amount of contact time in which they participate. The data failed to show any strong connection between students' total workload and grades they received; rather, they showed some evidence that regular presence at contact hours was the most positive influence on grades. Less frequent presence at lectures tended to indicate less time spent on independent study. It was also found that pre-clinical and clinical courses tended to require more work from students than other, more general subjects. While the present study does not provide conclusive evidence, it does indicate the need for further inquiry into the nature of the relationship between teaching and learning in higher education and for evaluation of the benefits (or otherwise) of more "self-directed" study.     

Influence of Random Factors and Two Different Staining Procedures on Computer-assisted Sperm Head Morphometry in Bulls

Contents Computer-assisted sperm analysis has the potential to improve reproducibility and objectivity in the assessment of sperm morphology. The aim of this study was to evaluate the use of a computer-assisted sperm morphometry analysis system for the determination of sperm head dimensions in bulls. Two experiments were performed to determine the variability caused by random factors and the influence of two different staining procedures. In the first experiment, three ejaculates were collected from each of five clinically healthy bulls. Air-dried semen smears were stained using a modified Farelly staining. The slides were observed via bright field microscopy with green filter using a 100× oil immersion objective. A video camera attached to the microscope transmitted images to a personal computer. Each sperm head was identified and analysed by the computer software (Morphology Analyzer V. 1.5; Mika Medical GmbH, Ismaning, Germany). Area, length and width of each sperm head were calculated and stored in a database for further statistical analysis. A minimum of 100 sperm heads were evaluated per slide. In experiment 2, the influence of two different staining procedures (Farelly and Papanicolaou) on sperm head dimensions was determined. The mean spermatozoal head measurements across all slides for area, length and width were 40.49 μm², 9.70 μm and 5.30 μm, respectively. On the basis of these results, the variability between slides, ejaculates and bulls using variance component estimation was calculated. All random factors (bull, ejaculate and slide) had a significant effect (p < 0.001) on sperm head dimensions. However, the variability attributable to bull (18.89–51.72%) was considerably higher compared with that of slide and ejaculate (0.17–5.27%). Additionally, differences existed between bulls concerning the shape and normality of histograms of their sperm head dimensions. The minimum number of spermatozoa required for analysis of sperm head dimensions was found to be about 60 spermatozoa per sample. The use of Papanicolaou stain resulted in significantly smaller sperm head dimensions, e.g. sperm head area 31.48 versus 38.35 μm² (p < 0.001). In conclusion, computer-assisted sperm head morphometry provides an objective, precise and reproducible tool. Comparisons of results from different studies should consider the influence of random and experimental factors to avoid misinterpretation.     

Evaluation of static telepathology in veterinary diagnostic cytology

BACKGROUND: Telepathology is the process of diagnostic pathology performed on digital images viewed on a display screen rather than by conventional light microscopy with glass slides. At present, electronic mail (e-mail) attachment is the most common communication medium for telepathology, because it is relatively simple and incurs minimal cost. OBJECTIVE: Our objective was to test whether telediagnosis through Internet e-mail could be applied to veterinary diagnostic cytology. METHODS: Using 20 cytology cases, on which a consensus diagnosis was reached by 3 experienced pathologists, a total of 130 images were digitized and sent as e-mail attachments from the Unit of Anatomic Pathology of the Faculty of Veterinary Medicine of Naples to the Unit of Pathology of the Faculty of Veterinary Medicine of Messina, Italy. The images were reviewed at the Unit of Pathology of Messina, where the consulting pathologist formulated his diagnoses by visualizing the images on the monitor of a computer. The telediagnoses were compared with the consensus diagnoses and with the consulting pathologist's interpretation using conventional light microscopy at a later date. RESULTS: The median time to capture images was approximately 30 minutes for each case. The median time to make a telecytodiagnosis was a few seconds for each case. Overall, there was good agreement (85%) between the consensus diagnosis and the consultant's telediagnosis. In 100% of the cases there was agreement between the consulting pathologist's telediagnosis and conventional glass slide diagnosis. CONCLUSION: Telepathology by e-mail provides acceptable efficacy and a faster turnaround time than post and can be applied to veterinary diagnostic cytology.     

Imaging of the lectin-labeled cell surface of human lymphocytes by the use of atomic force microscope

The atomic force microscope (AFM) is a new useful tool to examine the surface structure of specimens with a higher resolution than the conventional scanning electron microscope. In the present study, we used the AFM to observe the surface of paraformaldehyde-fixed human lymphocytes processed for histochemistry using a biotinylated lectin, wheat germ agglutinin, followed by colloidal gold and silver-enhancement method. Before the treatment, no particles were attached to the cell surface. After treatment, many particles about 100 to 150 nm in diameter were visualized on it. Since we could observe the same cells on the slide glass before and after treatment, the AFM has the advantage to enable us the repeated imaging of samples treated with various kinds of histochemistries.     

Evaluation of the expectations, learning and competencies of surgical skills by undergraduate veterinary students performing canine ovariohysterectomies

Objectives : To investigate the development of surgical skills of veterinary undergraduates and determine the number of canine ovariohysterectomies required to achieve competency and reduce levels of student concern. This was compared to student expectations and that of employers regarding surgical ability and provision of support to new graduates. Methods : A questionnaire regarding surgical concerns was sent to final year veterinary students enrolled within the University of Bristol, UK. A questionnaire was also sent to 200 UK veterinary practices regarding their impressions of surgical competence of new graduates and their provision of supervision. The responses were compared. Eleven additional final year students performed five canine ovariohysterectomies and graded their concerns. The number of supervised canine ovariohysterectomies required until competency was determined. Results : 80·4% of final year veterinary undergraduates replied that the surgical procedure which they were most concerned about their ability to perform was canine ovariohysterectomy. Students and veterinary practitioners differed in their opinions regarding whether they considered canine ovariohysterectomy to be a “day one skill” and what were desirable levels of supervision. Completing a minimum of four canine ovariohysterectomies led to 81·8% of students being assessed as competent. Clinical Significance : An unrealistically high expectation of competency by students may be a source of stress and concern. Employers should aim to provide hands‐on support whilst new graduates complete at least four canine ovariohysterectomies. Postoperative haemorrhage is uncommon but is the main concern for students.     

Implementing a multi-colour genetic marker analysis technique for embryology education

Embryology belongs to the basic sciences and is usually an integral part of the anatomy. The subject is traditionally taught by visual inspection of embryonic tissue slides stained with Haematoxylin and Eosin (H&E) to expose the dynamics of tissue histology as development proceeds. While combining in situ hybridization for gene expression analysis and immunostaining for protein expression analysis is an established technique for embryology research, the implementation of this tool in embryology teaching has not been described. The present study was conducted to assess the use of an online multi-colour gene expression analysis technique, alongside histological sections and diagrams, to improve students' understanding of embryology. The participants of this study were bachelor's students of Veterinary Medicine at the University of Khartoum. The method was also evaluated by distributing questionnaire items to Veterinary students via Google forms; subsequently, their responses were analysed qualitatively. The majority of students stated that the new technique was beneficial for their learning of embryology. The multi-colour images proved a more effective means for learning embryology than the traditional H&E image. Results from the students strengthen the belief in applying the multi-colour technique for better embryology course learning.     

APES处理的载玻片在原位杂交技术中的应用

目的：获得一种廉价的、核酸酶（RNase）处理效果好的载玻片运用于原位杂交技术中。方法：改进传统APES（3-Aminopropyl triethoxysilane）方法处理载玻片,加入载玻片高温烘烤、丙酮稀释APES等步骤,以去除RNase污染,并将处理好的载玻片分别进行冰冻和石蜡组织贴片,用原位杂交试验进行验证。结...     

Developments in undergraduate teaching of small-animal soft-tissue surgical skills at the University of Sydney

This article discusses recent developments in soft-tissue surgery teaching at the University of Sydney, Faculty of Veterinary Science. An integrated teaching program was developed for Bachelor of Veterinary Science (BVSc) students with the aim of providing them with optimal learning opportunities to meet "Day One" small-animal soft-tissue surgical competencies. Didactic lectures and tutorials were introduced earlier into the curriculum to prepare students for live-animal surgery practical. In addition to existing clinics, additional spay/neuter clinics were established in collaboration with animal welfare organizations to increase student exposure to live-animal surgery. A silicon-based, life-like canine ovariohysterectomy model was developed with the assistance of a model-making and special effects company. The model features elastic ovarian pedicles and suspensory ligaments, which can be stretched and broken like those of an actual dog. To monitor the volume and type of student surgical experience, an E-portfolio resource was established. This resource allows for the tracking of numbers of live, student-performed desexing surgeries and incorporates competency-based assessments and reflective tasks to be completed by students. Student feedback on the integrated surgical soft-tissue teaching program was assessed. Respondents were assessed in the fourth year of the degree and will have further opportunities to develop Day One small-animal soft-tissue surgical competencies in the fifth year. Ninety-four percent of respondents agreed or strongly agreed that they were motivated to participate in all aspects of the program, while 78% agreed or strongly agreed that they received an adequate opportunity to develop their skills and confidence in ovariohysterectomy or castration procedures through the fourth-year curriculum.     

THE NETWORK AND ITS ROLE IN DIGITAL IMAGING AND COMMUNICATIONS IN MEDICINE IMAGING

The elements of a digital imaging system are bound together by the network, so careful attention must be paid to this essential component. Networking hardware and cable choice will affect the speed of image transmission between devices within a network. Wireless networking offers convenience at the expense of speed and potentially, security. If a facility allows its network to connect to the Internet, security precautions are essential. Firewalls prevent unauthorized and destructive access to the network; virtual private networks allow encrypted communication with the network; and email and web browser encryption allow data transmitted from the network to other users on the Internet safely. This article presents an overview of this broad array of technologies. Readers are encouraged to seek additional depth as needed to address individual networking needs.