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Nanocomposites for Human Body Tissue Repair
Published in S. M. Sapuan, Y. Nukman, N. A. Abu Osman, R. A. Ilyas, Composites in Biomedical Applications, 2020
Cell attachment to a nanocomposite, nonporous or porous, is critical to all subsequent cellular activities. Therefore, adhesion between cells and nanocomposites should be examined. Confocal laser scanning microscopy (CLSM) is very useful for evaluating cell adhesion. Through immunofluorescence staining, vinculin antibody could be conjugated to the adhesive plaques and fluorescence signals can be used to locate where the adhesive plaques are formed (Wang et al., 2017). Besides, phalloidin and 4-6-diamidino-2-phenylindole (DAPI) can also be used in staining for visualizing the F-actin and nucleus of cells, assisting to investigate cell activities. Morphology of cells cultured on nanocomposites can be observed using SEM. In SEM sample preparation, gradient dehydration should be conducted first for cell-nanocomposite constructs, followed by critical-point drying. Before SEM observation, a thin layer of gold is coated on the dried cell-nanocomposite constructs. With cell culture experiments going for weeks and by examining the cell morphology at different culture times, the morphological changes of cells on nanocomposites could be observed (Huang et al., 1997; Tong et al., 2010).
Anammox process in a closed sponge-bed trickling filter
Published in Javier Adrián Sánchez Guillén, Autotrophic Nitrogen Removal from Low Concentrated Effluents, 2017
The total suspended solids (TSS), volatile suspended solids (VSS), soluble chemical oxygen demand (COD) and nitrite (NO2−-N) concentrations were determined according to Standard Methods for the Examination of Water and Wastewater (2012.). Ammonia (NH4+-N) was measured spectrophotometrically following the standard NEN 6472 (NEN, 1983). For nitrate (NO3−-N) measurements the standard ISO 7890-1:1986 (ISO 7890/1, 1986) was applied. pH was measured with a portable pH meter (Model ProfiLine 3310. WTW, Germany) and the dissolved oxygen (DO) concentrations were recorded using a Hach portable meter HQ30d equipped with the LDO101optical dissolved oxygen probe (Hach Company, USA). The average equivalent granule diameter of the Anammox inoculum was measured with a microscope Leica Microsystems M205 FA (magnification 13.0, calibration factor 4.45 and software version Qwin V3.5.1.; Leica Microsystems Ltd, Netherlands). Anammox bacteria were identified by Fluorescence in Situ Hybridization (FISH) technique. Combined biomass samples were scratched out from different sponges of each section and fixed in 4% (w/v) paraformaldehyde solution. Hybridization with fluorescent probes was performed as described by Schmid et al. (2000). Epifluorescence was used for cells identification and DAPI (4’, 6’-diamidino-2-phemylindol) as general DNA stain. Oligonucleotide probes were labeled with either fluorochromes Cy3 or Cy5 (Biomers.net, Germany). Images were acquainted by an epifluorescence microscope BX51 with a camera XM10 (Olympus, Japan). The probes used in this study were Amx820 (Schmid et al., 2001; Schmid et al., 2003), Amx1240 and Kst1275 (Schmid et al., 2005).
Fluorescence Microscopy
Published in Bethe A. Scalettar, James R. Abney, Cyan Cowap, Introductory Biomedical Imaging, 2022
Bethe A. Scalettar, James R. Abney, Cyan Cowap
Binding mechanisms are dye specific. DAPI binds to the minor groove of DNA with a resulting ∼20-fold enhancement in fluorescence. MitoTracker® dyes are cationic fluorophores that accumulate in active mitochondria. In some cases, small chemical fluorophores are membrane permeant and thus can be used to label living cells.
Is micronucleus assay in nasal mucosa cells an appropriate technique for detecting genotoxins by inhalation in humans? A systematic review
Published in International Journal of Environmental Health Research, 2023
Giovana Wagner Branda Drummond, Andrea Cristina de Moraes Malinverni, Ana Claudia Muniz Renno, Daniel Araki Ribeiro
The gold standard for staining was the DNA-specific technique with Feulgen-fast green, 4,6-diamidino-2-phenylindole dihydrochloride (DAPI), and acridine orange. In total, 11 studies met this requirement (Sarto et al. 1990; Ballarin et al. 1992; Burgaz et al. 2002; Huvinen et al. 2002; Godderis et al. 2004, Demircigil et al. 2010; Bruschweiler et al. 2014; Wultsch et al. 2015; Kesimci et al. 2017; Georg et al. 2019; Zeller et al. 2019). Two studies used other types of staining that were not appropriate for this review (Titenko-Holland et al. 1996; Ye et al. 2005). Almost all studies performed cell counting as the appropriate approach. Only two studies used different parameters to evaluate the incidence of micronuclei: scoring (Zeller et al. 2019) and UV excitation (Huvinen et al. 2002).
Effect of increasing salinity to adapted and non-adapted Anammox biofilms
Published in Environmental Technology, 2019
Steffen Engelbrecht, Mohammad Mozooni, Kristina Rathsack, Jörg Böllmann, Marion Martienssen
Fluorescence in situ hybridization (FISH) analysis was carried out as described by Amann et al. [27]. For detection of bacteria, probe EUB338 I (5′-GCT GCC TCC CGT AGG AGT-3′) [27] was applied. Three Anammox-specific probes were used, addressing the Anammox bacteria Candidatus K. stuttgartiensis (KST157; 5′-GTTCCGATTGCTCGAAAC-3′) [28], Candidatus Brocadia anammoxidans (Ban162; 5′-CGGTAGCCCCAATTGCTT-3′) [29] and Candidatus Brocadia fulgida (BFU613; 5`-GGATGCCGTTCTTCCGTTAAGCGG-3´) [30]. For nitrifying bacteria, the probes NSO1225 (5′-CGCCAT TGTATTACGTGTGA-3′), NSV443 (5′-CCG TGA CCG TTT CGT TCC G-3′), NSM156 (5′-TATTAGCACATC TTTCGAT-3′) (all modified from [31]) and NIT3 (5′-CCTGTGCTCCATGCTCCG-3′) (modified from [32]) were applied. DAPI (4′,6-Diamidin-2-phenylindol) was applied to stain the DNA. Image acquisition was performed using an epifluorescence microscope (Nikon Eclipse LV100 Pol, Nikon, Tokyo, Japan) and evaluated with the software NIS-Elements BR 3.10 (Nikon, Tokyo, Japan).
Nanofiltration fouling propensity caused by wastewater effluent organic matters and surface-water dissolved organic matters
Published in Environmental Technology, 2018
Wentao Shang, Feiyun Sun, Lichun Chen
The composition and spatial distribution of the microbial community of the biofouling layer on the NF surface over incubation time were examined under a fluorescent microscope after being stained. DAPI (4,6-diamidino-2-phenylindole) staining on all cells was performed following the method described by Sun et al. [16], who used a filtered DAPI solution (10 mg/mL in 25 mM Tris–HCl buffered saline, pH 7.0). For an NF membrane slice, it was dispersed by ultrasonic for 15 min to homogenize briefly the attached bacteria colony. The dispersed biomass was then air-dried on a slide and stained with the DAPI solution. After 10 min of staining, the slide was washed using a phosphate buffer saline solution and then air-dried at room temperature. The sample was examined under a fluorescent microscope (Eclipse, Nikon) with a 100 W high-pressure mercury lamp and a filter set MWU (Olympus Optical, excitation 330–385 nm).