Handling blood and body fluids in your clinical laboratory.
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Fourteen patients who had dermatitis herpetiformis of the skin with no gingival lesions were studied. In each patient biopsy specimens of normal gingiva, buccal mucosa, and perilesional skin were examined for IgA and IgG deposits. Depending on the method of handling biopsy specimens for immunofluorescence, IgA deposits were detected in 21 to 25 percent of gingival specimens, 27 to 46 percent of buccal mucosal specimens, and 64 to 76 percent of skin specimens. The occurrence of IgA deposits in the gingival papillae, a site where lesions of DH have not been reported in the past or observed in this study, suggests that these IgA deposits alone may not play a primary role in the pathogenesis of this disease. Similar immunofluorescent findings of IgA deposits were observed in both quick-frozen biopsy specimens and those placed in holding solution, indicating the suitability of the holding solution for transporting suggested DH biopsy material.
Differing incidences of Vibrio parahaemolyticus reported by separate government agencies are attributable to sample handling and subsequent isolation techniques.
The quality of biological samples is a major determinant of analytical reliability and translational relevance in patients with pancreatic ductal adenocarcinoma (PDAC). However, variability in specimen procurement, handling, transport, processing, and storage can substantially affect tissue integrity and the robustness of downstream analyses. This paper, promoted by the Pathology and Basic Science Task Force of the Italian Association for the Study of the Pancreas (AISP), brings together experts in pathology, molecular biology, translational research, medical oncology, and gastroenterology to provide practical recommendations for the collection, handling, and pre-analytical management of biological samples. Draft recommendations were discussed during dedicated working group meetings and approved by consensus among all authors, supported by key literature. The document identifies the biological specimen as the critical link between patient care, pathology, and research, and provides guidance for clinicians and professionals involved in sample procurement and processing. By addressing the requirements of different analytical platforms, including genomics, organoid generation, immunophenotyping, pharmacogenomics, and multiplex/spatial analyses, this paper aims to reduce pre-analytical variability, improve diagnostic accuracy, and enhance the clinical and translational value of molecular investigations in pancreatic cancer. Standardised procedures across centres may facilitate comparable data collection, support multicentre studies, and strengthen collaboration between clinicians, pathologists, and research laboratories.
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I describe the automation/mechanization of several areas of sample processing at SmithKline Beecham Clinical Laboratories. By analyzing and implementing new systems for conveyance and sorting of incoming specimens, specimen processing, within-laboratory specimen delivery, specimen storage and retrieval, and client resupply, we greatly improved the productivity of the nontechnical aspects of laboratory operations.
Twelve surgical specimens from radical vulvectomy and bilateral groin dissection were examined by xeroradiography to identify lymph nodes. Eight of the specimens were handled in a routine fashion, and four were chemically cleared. The xeroradiograph was not helpful in identifying small lymph nodes in the specimen, nor could any xeroradiographic characteristics specific for tumor-involved lymph nodes be identified.
Wild raptors brought into an ex situ environment often have poor semen quality that is further compromised by urine contamination. Generally, it is believed that in birds, artificial insemination into the cloaca or caudal vagina of females requires large doses of high-quality spermatozoa to maximize fertility. In an effort to define and overcome some of the challenges associated with reproduction in wild raptors, the objectives of this study were to 1) evaluate the frequency, impact, and remediation of urine contamination in fresh ejaculates for the purpose of maintaining sperm motility and viability in vitro, and 2) develop a deep insemination method that allows low numbers of washed sperm to be placed directly into the magnum to increase the probability of producing fertilized eggs. The species evaluated include golden eagle (Aquila chrysoetos), imperial eagle (A. adalberti), Bonelli's eagle (Hiernaetus fasciatus), and peregrine falcon (Falco peregrinus). Semen samples were collected and pooled by species, and a minimum of 25 pooled ejaculates per species were evaluated for urine contamination, pH, sperm viability, and sperm motility; the samples were either unwashed or washed in neutral (pH 7.0) or alkaline (pH 8.0) modified Lake's diluent. Female golden eagles and peregrine falcons were inseminated via transjunctional, intramagnal insemination with washed spermatozoa from urine-contaminated samples. Urine contamination occurred in 36.8 +/- 12.8% (mean +/- SEM) golden eagle, 43.1 +/- 9.1% imperial eagle. 28.7 +/- 16.1% Bonelli's eagle, and 48.2 +/- 17.3% peregrine falcon ejaculates. The pH in urine-contaminated semen samples ranged from 6.48 +/- 0.3 to 6.86 +/- 0.2, and in noncontaminated samples it ranged from from 7.17 +/- 0.1 to 7.56 +/- 0.1. Sperm viability and motility were reduced (P < 0.05) in all species for unwashed vs. washed sperm after 30 min incubation at room temperature. Two peregrine falcon chicks and one golden eagle chick hatched after intramagnal insemination. This study demonstrates that urine contamination, a common and lethal acidifier in manually collected raptor ejaculates, can be circumvented by immediate, gentle seminal washing. Furthermore, these processed sperm, when deposited by transjunctional intramagnal insemination, can produce live young.
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OBJECTIVES: Few studies have evaluated the handling methods used in the histological diagnosis of Helicobacter pylori. Filter paper has conventionally been used as a receptacle for the biopsy specimen before fixation. The aim of this study was to determine the presence of any effect caused by the use of filter paper. METHODS: The study population consisted of 104 consecutive patients undergoing endoscopic examination. Two antral biopsy specimens from the same area were obtained from each patient. One specimen was put onto a piece of filter paper, and the other into a plastic case. The specimens were fixed overnight in buffered formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin and Giemsa. A direct smear was also prepared from 77 patients by vigorously rubbing the filter paper on a glass slide and staining it with Giemsa. RESULTS: The detection rate of H. pylori was 47.1% (49 of 104) for the filter paper method, 56.7% (59 of 104) for the plastic case method, and 57.7% (60 of 104) for either of the two methods. Of the 60 positive patients, 11 filter paper specimens were negative, whereas only one plastic case specimen was negative. Statistical analysis revealed a significant difference between the two groups (p < 0.01). On the amount of H. pylori, the filter paper method showed a significantly lower grade than the plastic case method (p < 0.05). In the Giemsa-stained smears, H. pylori was identified in 17 (22.1%) of the 77 patients studied. CONCLUSIONS: Use of filter paper may decrease the sensitivity for detection of H. pylori infection. We recommend not using filter paper in the histological diagnosis of H. pylori.
Analysis of DNA and/or RNA is an important component of most epidemiologic studies. The methods used for their preparation vary depending on the number of samples in the study as well as the amount of tissue or cells available, the specific downstream assay, and the resources available. They range from classic phenol/chloroform extractions to robotic methods suitable for large-scale studies. Several methods are also available for the quantitation of DNA/RNA, including UV or fluorescence measurements and PCR. For DNA samples of limited quantity, it is also now possible to use whole genome amplification to increase the amount of DNA suitable for genotyping. Finally, a major issue in quality control is the careful labeling and handling of samples.