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Aetiology and Laboratory Diagnosis
Published in Raimo E Suhonen, Rodney P R Dawber, David H Ellis, Fungal Infections of the Skin, Hair and Nails, 2020
Raimo E Suhonen, Rodney P R Dawber, David H Ellis
This is a very rapid and sensitive method. However, a fluorescence microscope fitted with filters to give an excitation with ultraviolet light below 400 nra wavelength is required. Calcofluor white (M2R powder from Polysciences) or Blankophor BA (from Bayer) are used as whitening agents by the paper industry and selectively bind to cellulose and chitin. The dye fluoresces as it is exposed to ultraviolet light. Prepare two solutions: (1) dissolve 10 gm of KOH in 90 ml distilled water and then add 10 ml of glycerol; (2) dissolve 0.1 gm of calcofluor white powder in 100 ml of distilled water by gentle heating. Mix one drop of each solution on the centre of a clean microscope slide and mount a small portion of the specimen as described above. Gently heat the slide and examine microscopically for the presence of fungal elements that fluoresce a chalk-white or brilliant apple-green colour, depending on the filters used.
Animal Models for Studying Soft Tissue Biocompatibility of Biomaterials
Published in Yuehuei H. An, Richard J. Friedman, Animal Models in Orthopaedic Research, 2020
The equipment necessary for the sawing-grinding method (Exakt-cutting-grinding-system®, Exakt Apparatebau, Germany) consists of a precision guided, diamond-coated band saw and an automatic grinding machine. With the band saw, a first cut is made through the polymerized block. On the exposed tissue-implant surface, a microscope slide is glued. This slide is mounted again in the band saw using a vacuum slide holder. Subsequently, a planoparallel section is cut of a thickness between 50-200 μτη. This section is thinned down to a thickness of 5-10 fim with the grinding machine. Finally, the section is stained. For the staining, all the usually employed staining procedures for plastic embedded tissues can be used (see Chapter 7).
Aeroallergen sampling
Published in Richard F. Lockey, Dennis K. Ledford, Allergens and Allergen Immunotherapy, 2020
Estelle Levetin, Josh D. McLoud
A number of portable devices are designed to collect a single sample over a short period of time, typically from 5 to 15 minutes. The Burkard personal sampler (http://burkard.co.uk/) is a small cylindrical spore trap containing a pump within the unit (Figure 2.3). A 14 mm × 1 mm slit orifice is situated at the top of the sampler, and air is aspirated at 10 L/min with a particle cut point of 2.5 μm. Bioaerosols and other particulates are impacted as a single trace on a standard greased microscope slide. The Buck BioSlide (http://www.apbuck.com/shop/) also collects a single sample on a greased microscope slide. The flow rate of the sampling pump is adjustable from 10 to 20 L/min, and sampling time can be programmed for a 1-, 2-, 5-, or 10-minute collection period.
Baicalin attenuates XRCC1-mediated DNA repair to enhance the sensitivity of lung cancer cells to cisplatin
Published in Journal of Receptors and Signal Transduction, 2022
Zhangyong Yin, Enguo Chen, Xiaoping Cai, Enhui Gong, Yuling Li, Cunlai Xu, Zaiting Ye, Zhuo Cao, Jiongwei Pan
DNA damage for treated cells was assessed by comet assay based on the protocol from the previous report [18]. Briefly, treated cells (3 × 104) were used for this experiment. Normal melting agarose (1%) in PBS was dipped into a frosted microscope slide. Then, 10 μL cells blended with 90 μL 0.7% low melting agarose (37 °C) were added into pre-heat (37 °C) microscope slide. The microscope slide was immediately covered with a cover slip and kept at 4 °C for 10 min to allow the melting agarose to solidify. The cover slip was taken down and the microscope slide was treated with ice-cold lysis buffer for 2 h and kept shielded from light. Next, the microscope slide was placed in a horizontal electrophoresis tank and the cells were denatured in pre-cold alkaline buffer for 40 min. Then the cells were electrophoresed for 30 min at 25 V. The slides were washed three times by 0.4 mol/l Tris-HCl (pH 7.5), dyed by 20 μL ethidium bromide (5 μg/mL) for 10 min. The microscope slide was immediately covered with a cover slip, and cells were analyzed in each slide using a fluorescence microscope (Olympus BX 60 fluorescence microscope, Japan).
Protective effect of diphlorethohydroxycarmalol against oxidative stress-induced DNA damage and apoptosis in retinal pigment epithelial cells
Published in Cutaneous and Ocular Toxicology, 2019
Cheol Park, Hyesook Lee, Su Hyun Hong, Jeong-Hwan Kim, Seh-Kwang Park, Ji-Won Jeong, Gi-Young Kim, Jin Won Hyun, Seok Joong Yun, Byung Woo Kim, Wun-Jae Kim, Yung Hyun Choi
To measure the accumulation of intracellular ROS, ARPE19 cells were seeded onto 6-well plates with a density of 3 × 105 cells per well for 24 h, then treated either with or without DPHC (0, 25 and 50 μM) for 1 h, before adding H2O2 for a further 1 h. The cells were washed twice with cold phosphate buffered saline (PBS), suspended in PBS and stained with 10 µM DCF-DA for 20 min at 37 °C in the dark. The relative fluorescence intensities of the cell suspensions were measured using a flow cytometer (Becton Dickinson, San Jose, CA, USA). For image analysis for intracellular ROS production, the cells were seeded onto a coverslip-loaded 6-well plate. After 24 h of plating, cells were treated with DPHC (0, 25 and 50 μM), then 1 h later, 1 mM H2O2 was added to the plate for 1 h. After washing with PBS, 10 µM DCF-DA and 50 nM MitoTracker® Red were loaded in the well, then incubated at 37 °C for an additional 20 min. Next, the stained cells were washed and mounted on a microscope slide using a mounting medium. Finally, the images were visualized using a fluorescence microscope (Carl Zeiss, Oberkochen, Germany).
Malaria epidemiology and comparative reliability of diagnostic tools in Bannu; an endemic malaria focus in south of Khyber Pakhtunkhwa, Pakistan
Published in Pathogens and Global Health, 2019
Fatima Jahan, Nazma Habib Khan, Sobia Wahid, Zaki Ullah, Aisha Kausar, Naheed Ali
Finger-prick blood spots on filter paper from patients were processed with RDT and microscopy on-site (thick and thin smears) [26]. Briefly thin and thick slides were made on a clean, grease-free microscope slide and allowed to air dry. The films were stained with 10% Giemsa solution, allowed to air dry and then examined by microscopists at the labs/clinics with light microscopy using an oil immersion objective lens. A slide was declared negative only after observing 100 microscopic fields without finding parasites. Thick films were examined first for the detection of malaria parasites. Thick films in which malaria parasites were identified, were subsequently examined for species identification on their thin films as per standards described by WHO [27].