The primers used for chromatin immunoprecipitation followed by quantitative PCR (ChIP–qPCR) are listed in Table 3 . Briefly, CAL27 cells were grown to 85% confluence in a 150 mm culture dish (2 × 107 cells). After the media were removed, the cells were washed twice with 1× PBS and fixed with 0.75% formaldehyde (Sigma–Aldrich, Cat. No. 252549) for 10 min at room temperature. Glycine was added to a final concentration of 125 mM to stop cross‐linking, followed by a 5‐min incubation. The cells were washed twice with ice‐cold 1× PBS, scraped, and centrifuged for 5 min at 4°C at 1000g. ChIP lysis buffer (50 mM HEPES‐NaOH (pH 7.5), 140 mM NaCl, 1 mM EDTA, 1% Triton X‐100, 0.1% sodium deoxycholate, 0.1% SDS) supplemented with protease inhibitors was added to the pellet. The lysate was sonicated (VCX750, Sonics Inc., USA) for 8 min (1 s on/off cycles at 40% amplitude) and centrifuged at 8000g for 10 min at 4°C. Five percent of the total chromatin was used as an input. Immunoprecipitation was performed overnight with antibodies against H3K4me3 (Cell Signaling Technology, Cat. No. C42D8), H3K27ac (Cell Signaling Technology, Cat. No. D5E4), H3K27me3 (Cat. No. C36B11; Cell Signaling Technology, USA), and normal nonimmuno rabbit IgG (MerckMillipore, Cat. No. NI01) overnight with Protein A/G plus agarose beads (sc‐2003; Santa Cruz). The immunoprecipitated lysate was briefly centrifuged at 2000g for 1 min at 4°C, and the pellet was washed with high‐salt buffer (20 mM Tris‐HCl pH 8, 500 mM NaCl, 0.1% SDS, 1% Triton‐X‐100, 2 mM EDTA), low‐salt buffer (20 mM Tris‐HCl pH 8, 150 mM NaCl, 2 mM EDTA, 1% Triton‐X‐100, 0.1% SDS), and KCl buffer (10 mM Tris‐HCl pH 8, 1 mM EDTA, 1% sodium deoxycholate, 1% NP‐40, 0.25 M KCl). Reverse crosslinking was performed by adding TE (25 mM Tris–HCl, 1 mM EDTA) buffer supplemented with proteinase K (20 μg/mL) and RNase A (10 μg/mL) at 55°C for 15 min, followed by incubation at 100°C for 15 min. Approximately 500 pg of IP DNA was used for qPCR. The fold enrichment was calculated relative to the mock IgG control (nonimmune normal rabbit IgG), and the data are presented as an average of three biological replicates with standard deviation (SD).
Vcx 750
Manufactured by Sonics
388 citations
Sourced in United States, United Kingdom, Germany
About the product
The VCX 750 is a high-performance ultrasonic cell disruptor designed for efficient cell lysis and sample preparation. It utilizes advanced transducer technology to generate powerful ultrasonic waves, enabling rapid and effective disruption of cellular structures.
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Market Availability & Pricing
The VCX 750 ultrasonic processor is currently available from Sonics & Materials Inc. through authorized distributors. Prices for this model typically range from $5,757.69 to $7,962.97 based on information from secondary markets.
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Check compatibility
388 protocols using «vcx 750»
1
Chromatin Immunoprecipitation with Quantitative PCR
2
Microplastic-Induced Cytotoxicity Quantification
The effect of different concentrations of PS microplastics on LDH release was measured using the CyQUANT LDH Cytotoxicity assay kit (Thermo Fisher Scientific, USA) according to the manufacturer’s instruction. After incubation for 24 h, 50 μl of the supernatant was added to a 96-well plate. For maximum LDH release, 108 CFU/ml cells were lysed in an ice bath using a tip sonicator (VCX 750, Sonics & Materials, Inc., USA) at a duty cycle of 16.7% (10 s sonication and 50 s rest) for 20 min. The sonicated sample was added to a 96 well-plate, followed by addition of 50 μl of the substrate mixture and incubation without agitation at ambient temperature in the dark for 30 min. Then, 50 μl of the stop solution was added, absorbance was acquired with a spectrophotometer at 490 nm and 680 nm (background), and the difference in absorbance at these two wavelengths was determined. The LDH release was calculated using the LDH activity of the PS-treated sample (LDHT), spontaneous LDH activity (LDHS), and maximum LDH activity (LDHM):
The LDH release of each sample (LDHSample) was then expressed relative to the negative control (LDHNC) that had no microplastics:
The LDH release of each sample (LDHSample) was then expressed relative to the negative control (LDHNC) that had no microplastics:
3
MALDI-TOF MS Profiling of Mycobacterium abscessus
Mycobacterium abscessus isolates were cultured on blood agar (Clinag Co., Limited, Thailand) for five days at 37 °C. A full loop (5-10 mg) of the culture was then transferred to a microcentrifuge tube containing 400 µ L of TE buffer. This tube was heated at 95 °C for 30 minutes and then frozen at -20 °C for one hour. After thawing at room temperature, the sample was centrifuged at 13,000 rpm for 2 minutes. The supernatant was discarded, and 900 µ L of ethanol was added. The sample was centrifuged again at 13,000 rpm for 2 minutes, and the supernatant was removed. The cells were washed with 900 µ L of sterile water and ethanol, each time followed by centrifugation at 13,000 rpm for 2 minutes. After the washing steps, the supernatant was discarded, and the sample was allowed to dry at room temperature. Glass beads of the same volume as the pellet, 20 µ L of formic acid and 20 µ L of acetonitrile were added. The sample was homogenized with an Ultrasonic processor VCX-750 (Sonics, Sonics & Materials, Inc.) for 5 minutes and then centrifuged at 13,000 rpm for 10 minutes. Before analysis, 1 µ L of the supernatant was placed on a MALDI target plate and allowed to dry at room temperature. Then, 1 µ L of HCCA (α-cyano-4-hydroxycinnamic acid) was added to the MALDI target plate and also allowed to dry at room temperature. The MALDI target plate was analyzed using an auto-flex max within an hour.
4
Sonochemical Degradation of BPA under Various Conditions
Junsei Chemical Co. Ltd. (Tokyo, Japan) supplied potassium iodide (KI) and sodium sulfate (Na2SO4). Sigma-Aldrich Co. (USA) provided BPA (C15H16O2) and luminol (3-aminophthalhydrazide, C8H7N3O2). Samchun Pure Chemical Co., Ltd. (Korea) supplied sodium persulfate (Na2S2O8) and NaOH. All chemicals were used as received.
Three types of ultrasonic systems (20, 28, and 300 kHz) were used in this study, as shown inFig. 1 . All ultrasonic systems consisted of a circular glass vessel with a temperature-control double-layer wall, a recirculating chiller with a mixture of water and antifreeze (ethylene glycol), and an US generator [20 kHz probe-type sonicator with a replaceable tip (d: 13 mm) (VCX–750; Sonics & Materials Inc., USA), 28 kHz transducer module (Mirae Ultrasonic Tech., Bucheon, Korea), or 300 kHz transducer module (Mirae Ultrasonic Tech., Bucheon, Korea)]. Two double-layer wall vessels were used at 20 kHz and 28/300 kHz. The inner diameters of the glass vessels at 20 kHz and 28/300 kHz were 90 and 105 mm, respectively. The inner height of both vessels was 130 mm. A circular glass vessel without a double-layer bottom was mainly used for the 20 kHz systems in this study. A vessel with a double-layer wall and bottom was used to investigate the effect of the double-layer bottom on sonochemical oxidation activity, as shown in Fig. 1 . The electrical input powers, measured using a power meter (HPM-300A, ADPower, Korea), were fixed at 80 ± 2.1 W for all frequency conditions. The ultrasonic power, also known as calorimetric power, was calculated using Equation (2) : where Pcal is the ultrasonic or calorimetric energy, dT/dt is the rate of increase in liquid temperature, Cp is the specific heat capacity of the liquid (4.184 J/(g·K) for water), and M is the mass of the liquid. All prepared solutions for ultrasound irradiation in this study were air-saturated to ensure consistent results [52] (link), [53] (link), [54] (link), [55] (link), [56] (link).![]()
To determine the geometrically optimal probe position and liquid volume in the systems, KI dosimetry (KI conc.: 10 g/L) for the quantification of sonochemical oxidation activity and the sonochemiluminescence (SCL) method (0.1 g/L luminol and 1 g/L NaOH) for the visualization of the sonochemically active zone were used under various probe positions and liquid volumes.
It has been reported that the threshold for thermal activation of PS is around 40 ℃ [34] (link). So, six different temperature conditions (5–10 ℃, 15–20 ℃, 25–30 ℃, 45–50 ℃, 55–60 ℃, and no temperature control (NTC)) were applied in the only PS, only US, and US/PS processes in this study. The temperature in the liquid phase was measured every 15 min, and no considerable change in the controlled temperature range was observed. Under various temperature conditions, the BPA degradation tests [BPA conc.: 1.0 mg/L (0.004 mM)] were conducted in the only PS, only US, and US/PS processes at the determined optimal geometric conditions (Optimal liquid volumes of 400, 900, and 420 mL were determined for 20, 28, and 300 kHz, respectively, and this will be discussed in Chapter 3.1). The initial Na2S2O8 concentrations were 45 and 90 mg/L (0.19 and 0.38 mM, respectively). The degradation of BPA was analyzed using the pseudo 1st-order reaction kinetics.
The concentration of I3- ions, sonochemically generated in the KI solution, was measured using a UV–vis spectrophotometer (Libra S60; Biochrom Ltd., UK). BPA concentration was measured using a HPLC system (1260 Infinity II LC; Agilent, USA). The concentration of sulfate ions, evidence for PS activation, was quantified using ion chromatography (IC) system (ICS-2100; Dionex, USA). SCL images were obtained using a digital camera (DSC-RX100M7; Sony Corp., Japan) in a completely dark room (ISO-6400, F 2.8). The exposure time was 10–30 sec.
Three types of ultrasonic systems (20, 28, and 300 kHz) were used in this study, as shown in
Schematics of the 20 and 28/300 kHz ultrasonic systems and the temperature control recirculation system. A circular glass vessel without a double-layer bottom was mainly used for the 20 kHz systems. The image in the dotted line represents the vessel with a double-layer wall and bottom for comparison experiments.
To determine the geometrically optimal probe position and liquid volume in the systems, KI dosimetry (KI conc.: 10 g/L) for the quantification of sonochemical oxidation activity and the sonochemiluminescence (SCL) method (0.1 g/L luminol and 1 g/L NaOH) for the visualization of the sonochemically active zone were used under various probe positions and liquid volumes.
It has been reported that the threshold for thermal activation of PS is around 40 ℃ [34] (link). So, six different temperature conditions (5–10 ℃, 15–20 ℃, 25–30 ℃, 45–50 ℃, 55–60 ℃, and no temperature control (NTC)) were applied in the only PS, only US, and US/PS processes in this study. The temperature in the liquid phase was measured every 15 min, and no considerable change in the controlled temperature range was observed. Under various temperature conditions, the BPA degradation tests [BPA conc.: 1.0 mg/L (0.004 mM)] were conducted in the only PS, only US, and US/PS processes at the determined optimal geometric conditions (Optimal liquid volumes of 400, 900, and 420 mL were determined for 20, 28, and 300 kHz, respectively, and this will be discussed in Chapter 3.1). The initial Na2S2O8 concentrations were 45 and 90 mg/L (0.19 and 0.38 mM, respectively). The degradation of BPA was analyzed using the pseudo 1st-order reaction kinetics.
The concentration of I3- ions, sonochemically generated in the KI solution, was measured using a UV–vis spectrophotometer (Libra S60; Biochrom Ltd., UK). BPA concentration was measured using a HPLC system (1260 Infinity II LC; Agilent, USA). The concentration of sulfate ions, evidence for PS activation, was quantified using ion chromatography (IC) system (ICS-2100; Dionex, USA). SCL images were obtained using a digital camera (DSC-RX100M7; Sony Corp., Japan) in a completely dark room (ISO-6400, F 2.8). The exposure time was 10–30 sec.
5
Synthesis of Silica-TiO2 Nanocomposite
Silica nanostructures (60 mg) derived from Pennisetum glaucum residues and TiO2 nanoparticles (40 mg) were mixed with 200 mL of 5% gum arabic aqueous solution. Next, the mixture was sonicated at 750 W at 20 kHz for 30 min using a probe ultrasonicator (VCX-750, Sonics, Newtown, CT, USA). The obtained colloid was dried at 60 °C. Following that, the obtained material was used for further studies.
Top 5 most cited protocols using «vcx 750»
1
Quantification of Extracellular Polymeric Substances in Planktonic and Biofilm Cultures
The EPS was extracted using a modification of the sonication method described previously [45 (link),46 ]. This modification was based on the work of Vandevivere and Kirchman [47 (link)]. Briefly, planktonic samples were prepared using 5 mL overnight culture of PA14 cells (OD at 595 nm, ~1.5) with appropriate concentrations of ginger (1~10%). The cultured cells were then harvested via centrifugation at 8,000 × g and resuspended in 10 mL 0.01 M KCl. Biofilm samples, on the other hand, were prepared by also using the overnight culture. The aliquot was diluted with fresh AB medium (1:20) with appropriate amounts of ginger extract. Then, 3 ml of the dilution were aliquoted into borosilicate bottles and incubated at 37°C for 24 h without agitation. The suspended cultures were measured by spectrophotometer at OD 595 nm then discarded. The bottles were washed with phosphate-buffered saline (pH = 7.2) to remove any remaining suspended cells. Biofilm cells on the wall were removed by vortexing and scraping after addition of 3 ml 0.01 M KCl. The next steps in processing were the same for both planktonic and biofilm cells and are as follows. The cells were disrupted with a sonicator (VCX 750, SONICS, Newtown, CT, USA) for 4 cycles of 5 s of operation and 5 s of pause at a power level of 3.5 Hz. The sonication method did not result in significant cell lysis. This was confirmed by conducting a cell counting experiment in which the number of dead cells after sonication was less than 10% (Figure S1 ). The sonicated suspension was centrifuged (4,000 × g, 20 min, 4°C), and the supernatant was then filtered through a 0.22-µm membrane filter (Millex® filter, Carl Roth). The amounts of protein and carbohydrate in the filtrate were analyzed. For the analysis of protein, 40µL filtrate was aliquoted onto a 96-well polystyrene microtiter plate, and 200 µL Lowry reagent (L3540, Sigma Aldrich, St. Louis, Missouri, USA) was added to the aliquots. After 10 min of incubation at room temperature, 20 µL Folin-Ciocalteu reagent (L3540, Sigma Aldrich) was added to the mixture. After another 30 min of incubation at room temperature, absorbance at 750 nm was measured using the iMark microplate reader. The amount of protein was quantified by dividing OD at 750 nm by OD at 595 nm.
Carbohydrate was analyzed in a manner similar to that used to quantify protein. Fifty microliters of the filtrate were aliquoted in a 96-well polystyrene microtiter plate, and 150 µL 99.9% sulfuric acid was added to the aliquots. After 30 min of incubation at room temperature, 5% phenol was added to the mixture. After another 5 min of incubation at 90°C in the water bath, absorbance at 490 nm was measured using the iMark microplate reader. The amount of carbohydrate was quantified by dividing OD at 490 nm by OD at 595 nm.
Carbohydrate was analyzed in a manner similar to that used to quantify protein. Fifty microliters of the filtrate were aliquoted in a 96-well polystyrene microtiter plate, and 150 µL 99.9% sulfuric acid was added to the aliquots. After 30 min of incubation at room temperature, 5% phenol was added to the mixture. After another 5 min of incubation at 90°C in the water bath, absorbance at 490 nm was measured using the iMark microplate reader. The amount of carbohydrate was quantified by dividing OD at 490 nm by OD at 595 nm.
Corresponding organizations : Korea University
2
Synthesis of mPEGylated Lipid-based Micelles
The DTX-loaded LsbMDDs was prepared following previously reported procedures with minor modifications to introduce mPEGylation with DSPE-PEG2K or DSPE-PEG5K into the outer shell portion of micelles (Chen et al., 2015 (link); Su et al., 2018 (link)). In brief, a fixed drug (DTX)/amphiphilic polymer (DSPE-PEG2K) ratio of 1:5 with the addition of an appropriate amount of TPGS as an antioxidant was first dissolved in methanol, and a thin film was formed after evaporation (Rotavapor R124; Buchi, Flawil, Switzerland) of the organic solvent. Soybean lecithin (S100) at 1000 and 375 mg of DSPE-PEG5K (or DSPE-PEG2K) were suspended in 25 mL of deionized water and then subjected to ultrasonication (VCX 750, 20 kHz, Sonics and Materials, Market Harborough, United Kingdom) to form a lecithin/DSPE-PEG (2K or 5K) nanosuspension. Then 1 mL of the lecithin/DSPE-PEG (2K or 5K) nanosuspension was used to hydrate the thin film obtained above, and the reconstituted mixture was further subjected to ultrasonication at full power for at least 5 min while maintaining a constant temperature to form lecithin-stabilized NCs in the solution. Any unencapsulated drug was discarded by filtering this NC solution through a 0.22-μm membrane (Millipore, Billerica, MA). An appropriate amount of an anti-freeze agent was added to the filtrate and then freeze-dried to obtain the dry powder form of NCs. Two amphiphilic polymers of DSPE-PEG2K and DSPE-PEG5K with different PEG chain lengths were used to formulate the DTX-loaded mPEGylated LsbMDDs, respectively, designated DTX-loaded LsbMDDs(2K) and DTX-loaded LsbMDDs(5K). To prepare the DIO (green fluorescence for in vitro assays)-loaded or DIR (near-infrared fluorescence for in vivo imaging)-loaded LsbMDDs, we followed the same procedure as that for the DTX-loaded LsbMDDs except that the DTX was replaced with either DIO or DIR.
Corresponding organizations : Taipei Medical University, Yuanpei University, Taipei Medical University Hospital
3
Ultrasound-Assisted Nanoemulsion Formulation
Nanoemulsions were prepared by ultrasound treatment with EPS as an emulsifier, PEG400 as a co-emulsifier, liquid paraffin oil as an oil phase, and NYS as the drug. EPS was dissolved in distilled water, then PEG400 was added and vortexed (QL-901, China, Aomen) for 5 min. NYS was weighed and dissolved in liquid paraffin oil, and added into the EPS solution and vortexed for 5 min to obtain raw EPS/NYS emulsions. The emulsions were ultrasonicated (SONICS, VCX750) using a 3 mm diameter probe and an interval time of 5 s on, 5 s off. Throughout the ultrasound treatment, the container containing the sample was in an ice bath, and temperature difference in the system was less than 20°C.
Corresponding organizations : Weifang Medical University
4
Dual-Drug Loaded PLGA Nanoparticles
Drug-loaded PLGA nanoparticles were prepared by the solvent evaporation method according to previous reports,36 (link)–39 (link) with slight modification. The organic phase of quercetin 0.05 g, voglibose 0.01 g, and PLGA 0.25 g solubilized in 3 mL of acetone was added dropwise into 30 mL of the aqueous phase containing 0.194 g of Tween-20 (as emulsifier) under magnetic stirring at 2500 rpm for one hour to generate a PLGA-drug nanoemulsion. Subsequently, oleic acid (at ratios of 4%, 4.5%, 5%, 5.5%, and 6% w/v) was added to the crude emulsion, which was then sonicated for 5 minutes with a probe sonicator (VCX750; Sonics and Materials Inc, Danbury, CT) at an optimal amplitude of 40% and a frequency of 20 kHz. Sonication is a crucial step in preparation of a nanoemulsion, and can increase the temperature to that needed to inactivate antioxidants. Therefore, we utilized an on/off cycle to maintain a low temperature. The solution gradually changed from clear light yellow to milk-white yellow. Oleic acid acted as both a chemical permeation enhancer and as a surfactant to obtain a homogeneous nanoemulsion.25 (link),40 The nanoemulsion was diluted by adding water to make a final formulation of 70 mL. The acetone was removed, and the aqueous phase was concentrated by rotary evaporation at 40°C under reduced pressure. The concentrated nanoemulsion contained nanoparticles of the smallest size. These nanoparticles were centrifuged, washed, freeze-dried, and stored at room temperature for further experiments.
Corresponding organizations : Gachon University
5
Scalable Giant Vesicle Production
The w/o emulsion transfer method was chosen to produce GVs as previously described [15 (link)] with some minor modifications for some lipids. Calcein was added to the inner aqueous solution enabling quantitative analysis of GVs by flow cytometry. After the lipids were dissolved and the chloroform was entirely evaporated, the w/o emulsion was made by sonication using an ultrasonic processor (SONICS & MATERIALS, INC, VCX750, Newton, Connecticut, USA) on ice for 1 min (Ampl. 40%, pulse on/off 2 s/1 s) and equilibrated on ice for 10 min. Considering that lipid species used in this study have different structures and transition temperatures, the sonication conditions for the emulsion produced with those lipids were optimized, as listed in Table 2 . The bottom aqueous solution was placed in a new microcentrifuge tube, and the w/o emulsion obtained in the previous step was gently poured on top of the bottom aqueous solution. After standing for 5−10 min to equilibrate the interface, the emulsion was centrifuged to form a pellet of GVs. The supernatant oil was removed and a fresh bottom aqueous solution was added to resuspend the pellet. Oil traces were removed by another centrifugation round. Finally, 100 μL of this GV suspension was prepared and immediately analyzed by IFC or stored at 4 °C for further stability analysis.
Corresponding organizations : East China Normal University
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