Thirty-five μl of purified VLPs were diluted in 10 ml SM buffer and filtered onto a 0.02-μm Anodisc polycarbonate filter (Whatman, Maidstone, UK). Filters were stained with 2×SYBR Gold (Thermo Fisher Scientific) for 15 min, then washed with H2O once. After drying, the filter was mounted on a glass slide with 15 μl of mountant buffer (100ul 10% ascorbic acid + 4.9ml 7.4 PBS + 5ml 100% glycerol; filtered through 0.02um). For each filter, viruses were counted in 5 to 10 fields of view selected randomly. The filter was visualized using a motorized inverted system microscope IX81 (Shinjuku, Tokyo, Japan) for fluorescence. VLPs were counted using imageJ. Stained particles <0.5 μm in diameter were regarded as VLPs (larger particles were not counted). Purified lambda phage with known plaque-forming unit counts (PFU) per ml were used as a positive control to adjust image color, saturation, level and contrast. VLPs mock purified from SM buffer was used as negative controls. At least one count per microscope field was set as a threshold for a positive detection, which was equal to ~6.6 ×106 counts per gram feces. Lower than this threshold, the VLP counts were considered to be below the level of detection. The results were listed in Supplementary Table 2 .
Anodisc polycarbonate filter
Manufactured by Cytiva
Sourced in United Kingdom
The Anodisc polycarbonate filter is a laboratory instrument used for the filtration of samples. It is designed to remove particulates from liquids or gases by passing them through a porous membrane. The filter is made of polycarbonate material and features a uniform pore size to ensure consistent and reliable filtration performance.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using anodisc polycarbonate filter
1
Quantification of Virus-Like Particles in Feces
2
Quantification of Microbes in Eel Aquaculture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples from surrounding water and epidermal mucosa of eels farmed in tanks at 22°C and 1% salinity in facilities at University of Valencia (Planta de Acuarios de Experimentación, PAE) were collected. Samples were maintained on ice, sonicated in 3 pulses during 4 s. 50 and 3 ml of water and mucus, respectively, were directly filtered per 0.02 μm Anodisc polycarbonate filter (Whatman). Anodisc filters were stained with SYBR Green 5x, washed and visualized using epifluorescence microscope. For each sample, 25–30 images were analyzed using ImageJ (Schneider et al., 2012 (link)). Counts of bacteria and virus-like particles per milliliter were made using a previous protocol described (Patel et al., 2007 (link)).
3
Quantification of Virus-Like Particles in Feces
4
Viral Particle Enumeration in Fecal Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VLPs were enumerated as described previously (17 (link)). Briefly, a 0.1-g sample of the rectal luminal content was suspended in 10 ml of sterilized saline magnesium buffer (SM buffer; 100 mM NaCl, 8 mM MgSO4, 50 mM Tris-HCl [pH 7.4], and 0.002% gelatin; filtered through a 0.02-μm Anodisc polycarbonate filter [Whatman] before use). After serial filtration through 5-, 0.45-, and 0.2-μm pore size syringe filters (Sartorius), the filtrate was serially diluted 10-fold, and the same dilutions of the normal and diarrheic samples were compared. The filtrates were then filtered through a 0.02-μm Anodisc filter. The filters were stained with 5× SYBR gold for DNA viruses for 10 min, washed once, and visualized under an Eclipse 50i microscope (Nikon) equipped with an Intensilight C-HGFI device (Nikon). VLP images (×1,000 magnification) were obtained, 10 images from different fields of view per sample, and VLPs were counted using an i-Solution image analyzer (InnerView, Seoul, South Korea). The SM buffer was used as a negative control.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!