The membrane-disruptive properties of the LTX-315 peptide were characterized by using a Q-Sense E4 instrument (Biolin Scientific AB, Gothenburg, Sweden), as previously described [37 (link)]. Silica- and titania-coated QCM-D sensor chips (model nos. QSX303 and QSX310, Biolin Scientific AB) were used for experiments involving SLB and intact vesicle platforms, respectively. Before experiment, the sensor chips were rinsed with 1% (wt/v) sodium dodecyl sulfate (SDS), water, and ethanol sequentially. After drying with a stream of nitrogen gas, each sensor chip was treated with oxygen plasma for ~1 min by using a CUTE-1MPR machine (Femto Science Inc., Hwaseong, Republic of Korea). Afterwards, the sensor chips were mounted in the measurement chambers and signal baselines were first establishing by injecting buffer solution into the measurement chambers. All liquid samples were introduced by using a Reglo Digital MS-4/6 peristaltic pump (Ismatec, Glattsburg, Switzerland) with a volumetric flow rate of 50 µL·min−1. The temperature in each QCM-D measurement chamber was set at 25 °C during experiment, and the resonance frequency (Δf) and energy dissipation (ΔD) shifts were monitored as a function of time. Measurement data were collected at several odd overtones (n = 3–13), and the presented data are reported from the fifth overtone (n = 5) and normalized according to the overtone number.
Q sense e4 instrument
Manufactured by Biolin Scientific
Sourced in Sweden
The Q-Sense E4 instrument is a versatile tool for studying surface interactions and material behavior. It measures changes in the resonance frequency and dissipation of a vibrating quartz crystal sensor, which can provide insights into surface phenomena such as adsorption, thin film formation, and viscoelastic properties of materials. The instrument allows for real-time monitoring of these processes under controlled environmental conditions.
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Lab products found in correlation
Gold coated qcm d sensors, by Biolin Scientific (2 mentions)
Qsense dfind, by Biolin Scientific (2 mentions)
Ne 1000 syringe pump, by New Era Pump Systems (1 mentions)
Microfuge 16, by Beckman Coulter (1 mentions)
V 660 spectrometer, by Jasco (1 mentions)
Liposofast liposome extruder, by Avestin (1 mentions)
Pbs ph 7, by Merck Group (1 mentions)
Neutravidin, by Thermo Fisher Scientific (1 mentions)
10 protocols using q sense e4 instrument
1
Characterization of LTX-315 Peptide Membrane Disruption
2
Quartz Crystal Microbalance Study of TiO2 Surfaces
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A Q-Sense E4 instrument (Biolin Scientific AB, Stockholm, Sweden) was used for all QCM-D experiments involving deposition kinetics and interaction processes. Liquid samples were injected into the measurement chamber by using a Reglo Digital MS-4/6 peristaltic pump (Ismatec, Glattsburg, Switzerland), and the flow rate was 50 µL/min. TiO2-coated QCM-D sensor chips (product no.: QSX310, Biolin Scientific AB, Gothenburg, Sweden) were used for experiments, and the main TiO2 phase on the sensor surface was anatase along with titanium [38 ,39 (link)]. Prior to the experiment, the sensor chips were sequentially rinsed with 1% (w/v) aqueous sodium dodecyl sulfate, deionized water, and ethanol, and then dried under a gentle stream of nitrogen gas. The rinsed chips were then treated with oxygen plasma for 2 min in a CUTE-1MPR oxygen plasma chamber (Femto Science Inc., Hwaseong, Korea) before being mounted in the QCM-D measurement chambers. In each experiment, a stable baseline signal was first established in aqueous buffer and the reported data were collected at the 5th odd overtone. In applicable cases, the Sauerbrey equation [40 (link)] was applied to calculate the surface mass density of adsorbed molecules, and the mass sensitivity constant used in the calculations was 17.7 ng/cm2 per 1 Hz shift.
3
Protein Adsorption Quantification by QCM-D
4
Quantifying Protein Adsorption on Dhvar5-Chitosan
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Protein adsorption to Dhvar5-chitosan coatings was evaluated using a QCM-D system (Q-Sense E4 instrument; Biolin Scientific, Gothenburg, Sweden) and gold-coated QCM-D sensors with a fundamental frequency of 5 MHz (Biolin Scientific), as previously described [35 (link)]. Briefly, after cleaning, the sensors were prepared as described above (Section 2.2 ) [14 (link)]. Adsorption of bovine serum albumin (BSA) to Dhvar5-Chitosan surfaces was followed in real-time by the frequency and dissipation shifts of the sensors. A baseline was established by pre-incubating samples with phosphate-buffered saline (PBS, pH = 7.4) for 15 min at a flow rate of 0.1 mL/min. Then, a BSA solution at 4 mg/mL in PBS was injected into the system at a flow rate of 25 µL/min until saturation was achieved (2 h). PBS was then used to remove loosely attached proteins. The temperature was kept at 37 °C throughout the assay. The resulting data was treated with the Voigt model. Three replicates per sample were analyzed in three independent assays (n = 3).
5
Lysozyme Adsorption on BCP40 Mesoporous Films
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Enzyme storage was studied with a quartz
crystal microbalance (Q-Sense E4 instrument, Biolin Scientific) on
BCP40 mesoporous films prepared onto silica-coated QCM
sensors (5 MHz 14 mm Cr/Au/SiO2, Quartz PRO) with an active
area of 0.79 cm2. In a continuous measurement, lysozyme
adsorption was induced by pumping 2 mg mL–1 of lysozyme
in 0.1 M PBS buffer (pH 7.3) into the QCM chamber, and subsequent
lysozyme desorption was induced by pumping PBS buffer into the chamber.
Lysozyme and PBS were pumped into the QCM chamber at a flow rate of
30 μL min–1. Frequency analysis, conversion
to the Sauerbrey mass using the composite Sauerbrey of the frequency
harmonics f3, f5, f7, f9, f11, and f13, and
validation of the model were performed with the software QSense Dfind
(Biolin Scientific).
crystal microbalance (Q-Sense E4 instrument, Biolin Scientific) on
BCP40 mesoporous films prepared onto silica-coated QCM
sensors (5 MHz 14 mm Cr/Au/SiO2, Quartz PRO) with an active
area of 0.79 cm2. In a continuous measurement, lysozyme
adsorption was induced by pumping 2 mg mL–1 of lysozyme
in 0.1 M PBS buffer (pH 7.3) into the QCM chamber, and subsequent
lysozyme desorption was induced by pumping PBS buffer into the chamber.
Lysozyme and PBS were pumped into the QCM chamber at a flow rate of
30 μL min–1. Frequency analysis, conversion
to the Sauerbrey mass using the composite Sauerbrey of the frequency
harmonics f3, f5, f7, f9, f11, and f13, and
validation of the model were performed with the software QSense Dfind
(Biolin Scientific).
6
Quartz Crystal Microbalance with Dissipation Measurements
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QMCD measurements were
performed with a Q-Sense E4 instrument (Biolin Scientific) and a NE-1000
syringe pump (New Era pump systems) for flow control. All measurements
were performed at a set temperature of 25 °C at a flow rate of
100 μL/min. Voight modeling21 (link) and
curve fitting was performed with the instrument specific software
package Qtools using overtones 3, 5, 7, and 9. To obtain frequency
and dissipation signals relative to a blank QCMD crystal, the absolute
frequency and dissipation were recorded in the same liquid environment
prior to PEG grafting.
performed with a Q-Sense E4 instrument (Biolin Scientific) and a NE-1000
syringe pump (New Era pump systems) for flow control. All measurements
were performed at a set temperature of 25 °C at a flow rate of
100 μL/min. Voight modeling21 (link) and
curve fitting was performed with the instrument specific software
package Qtools using overtones 3, 5, 7, and 9. To obtain frequency
and dissipation signals relative to a blank QCMD crystal, the absolute
frequency and dissipation were recorded in the same liquid environment
prior to PEG grafting.
7
Enzyme Immobilization Quantified by QCM
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Enzyme immobilization into
the porous aluminosilicate coating was studied with a quartz crystal
microbalance with dissipation monitoring (Q-Sense E4 instrument, Biolin
Scientific) using a previously coated QCM sensor (5 MHz, 14 mm Cr/Au/SiO2, 0.79 cm2 active area, Quartz PRO). Solutions
were pumped at a flow rate of 30 μL min–1 into
the QCM chamber. QCM analysis (frequency and dissipation) of the harmonics f3, f5, f7, f9, f11, and f13 was performed with
the software QSense Dfind (Biolin Scientific).
the porous aluminosilicate coating was studied with a quartz crystal
microbalance with dissipation monitoring (Q-Sense E4 instrument, Biolin
Scientific) using a previously coated QCM sensor (5 MHz, 14 mm Cr/Au/SiO2, 0.79 cm2 active area, Quartz PRO). Solutions
were pumped at a flow rate of 30 μL min–1 into
the QCM chamber. QCM analysis (frequency and dissipation) of the harmonics f3, f5, f7, f9, f11, and f13 was performed with
the software QSense Dfind (Biolin Scientific).
8
Biophysical Characterization of Nanoparticles
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Fluorescence measurements were recorded using a Perkin Elmer LS55 fluorimeter, UV-visible spectrophotometry was performed on a V-660 spectrometer (Jasco (UK) Ltd). Vesicles were extruded using a LiposoFast liposome extruder (AVESTIN Europe GmbH). Sonication was performed using a VCX130PB probe-type sonicator (Sonics & Materials Inc., 130 W power rating). Centrifugation was performed using a Heraeus Instruments Megafuge 1.0R for large tubes and a Beckman Coulter Microfuge 16 for Eppendorf tubes. Vortex mixing was performed using a Vortex Genie 2 (600-2700 rpm). Transmission electron microscopy (TEM) was performed using a Jeol 1220, 120 kV instrument with a GATAN ORIUS CCD camera or a Tecnai T20, 220 kV. Inductive heating of nanoparticles was conducted using a water-cooled EASYHEAT 0224 induction heater (2.0 kW, 150-400 kHz) with an EASYHEAT 300P workhead (392 kHz, 3 cm loop diameter, 1.5 turns). QCM-D was performed using a Q-Sense E4 instrument (Biolin Scientific, UK). NdFeB magnets ring/cylinder shape (5600 Gauss) were obtained from Magnet Expert Ltd (UK).
9
Acoustic Sensing of Biomolecular Interactions
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2.1 Acoustic array and QCM sensor. The 150 MHz HFF-QCM acoustic array signals were monitored with a newly developed platform (AWS, S.L. Paterna, Spain). The QCM sensors (AWS, S.L. Paterna, Spain), with a fundamental frequency of 5 MHz, were monitored using the Q-Sense E4 instrument (Biolin Scientific, Sweden) at the 7th overtone (35MHz). Details on the device cleaning can be found in the Supplementary S1. 2.2 Acoustic detection of b-BSA and NAv. Protein samples were diluted in PBS pH=7.4 (Sigma-Aldrich). Neutravidin (NAv-Invitrogen) (0.2 mg/mL) and biotinylated-BSA (b-BSA) (0.2 mg/mL) were applied on the device surface; 0.05 mg/mL of NAv was further applied on the b-BSA layer. b-BSA was prepared as described in SI-S2. Due to the different size of the HFF QCM and QCM devices and fluidics geometry, the working volumes were V150MHz=60 μL and V35MHz=200 μL. 2.3 HFF QCM detection of dsDNA and liposomes. dsDNA fragments of 21 bp, 50 bp 75 bp and 157 bp were prepared according to 24 and applied (60 μL of 83 or 500 nM) to a NAv pre-coated array. 100 μL x 0.2 mg/mL of 200 nm of POPC liposomes (1-palmitoyl-2-oleoyl-glycero-3-phosphocholine), prepared by extrusion as described before 24 , were added on the DNA surface.
10
Laminin adsorption on QCM-D sensors
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Gold-coated QCM-D sensors (Biolin Scientific) were cleaned as previously described [41] . Clean sensors were further modified following the procedure reported in 2.8. A QCM-D system (Q-Sense E4 instrument, Biolin Scientific) was used to monitor in real time the frequency (Δf) and dissipation Version: Postprint (identical content as published paper) This is a self-archived document from i3S -Instituto de Investigação e Inovação em Saúde in the University of Porto Open Repository For Open Access to more of our publications, please visit http://repositorio-aberto.up.pt/ A01/00 (ΔD) shifts related to laminin adsorption as described in Supplementary Materials and Methods. Data was modeled using the Voigt model [41] in the QTools® V3 software. Results are presented as mass per surface area (ng/cm2).
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