Before electroporation, Jurkat cells were harvested by centrifugation (10 min, 200× g, 4 °C) and resuspended in an appropriate electroporation medium. The cell concentration was determined using a Coulter counter (Z2, Beckman Coulter, Brea, CA, USA). The cell suspension (3.0 μL) containing 1.0 × 105 Jurkat cells was dispensed in the silicone oil, and a high voltage was supplied using a DC high-voltage (HV) power supply (HAR-30R10, Matsusada Precision, Kusatsu, Japan). When the applied voltage was set to 3.0 kV, instantaneous short-circuiting could be induced. After short-circuiting, with a distinctive sound, the DC HV power supply was manually turned off.
Kf 96 100cs
Manufactured by Shin-Etsu Chemical
Sourced in Japan
KF-96-100CS is a silicone fluid product manufactured by Shin-Etsu Chemical. It is a clear, colorless liquid with a viscosity of 100 centistokes. The product is designed for use as a lubricant, damping fluid, and heat transfer medium in various industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Mh2 500n, by Imada (3 mentions)
Fm4 64, by Thermo Fisher Scientific (3 mentions)
Fluorinert, by 3M (1 mentions)
Aniline blue, by Merck Group (1 mentions)
Axiocam 506 color, by Zeiss (1 mentions)
Axio imager a2 upright microscope, by Zeiss (1 mentions)
Congo red, by Fujifilm (1 mentions)
X smart plus endodontic motor, by Dentsply (1 mentions)
X smart plus, by Dentsply (1 mentions)
6 protocols using kf 96 100cs
1
Electroporation of Jurkat Cells in Silicone Oil
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Figure 1 shows the experimental setup. The apparatus was fabricated with pin headers and a printed circuit board as described in the previous paper [35 (link)]. The electrodes’ gap was 5.08 mm. The apparatus was filled with 1.5 mL fluorocarbon oil (Fluorinert, 3M, Tokyo, Japan), and then 1 mL silicone oil (KF96-100CS, 100 cSt kinematic viscosity, 2.74 relative dielectric constant, 965 kg/m3 density, Shin-Etsu Chemical, Tokyo, Japan) was added.
Before electroporation, Jurkat cells were harvested by centrifugation (10 min, 200× g, 4 °C) and resuspended in an appropriate electroporation medium. The cell concentration was determined using a Coulter counter (Z2, Beckman Coulter, Brea, CA, USA). The cell suspension (3.0 μL) containing 1.0 × 105 Jurkat cells was dispensed in the silicone oil, and a high voltage was supplied using a DC high-voltage (HV) power supply (HAR-30R10, Matsusada Precision, Kusatsu, Japan). When the applied voltage was set to 3.0 kV, instantaneous short-circuiting could be induced. After short-circuiting, with a distinctive sound, the DC HV power supply was manually turned off.
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Before electroporation, Jurkat cells were harvested by centrifugation (10 min, 200× g, 4 °C) and resuspended in an appropriate electroporation medium. The cell concentration was determined using a Coulter counter (Z2, Beckman Coulter, Brea, CA, USA). The cell suspension (3.0 μL) containing 1.0 × 105 Jurkat cells was dispensed in the silicone oil, and a high voltage was supplied using a DC high-voltage (HV) power supply (HAR-30R10, Matsusada Precision, Kusatsu, Japan). When the applied voltage was set to 3.0 kV, instantaneous short-circuiting could be induced. After short-circuiting, with a distinctive sound, the DC HV power supply was manually turned off.
2
Fluorescent Staining of Pollen Tubes
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For evaluation of fluorescent staining of pollen tubes on the medium and in the micropyle, aniline blue (415049; Merck, Germany), fluorescent brightener 28 (used as calcofluor white; F3543; Merck, Germany), Congo red (032-03922; Wako, Japan), PI (P4864; Merck, Germany), FM4-64 (F34653; Thermo Fisher Scientific, Waltham, MA, USA), and FDA (F7378; Merck, Germany) were prepared in the pollen germination medium without agarose. The final concentration of each dye is shown in Figure 1 . At 5.5 h after pollination, 20 µL of each dye was dropped on the medium. After 5 min, staining of pollen tubes was observed with an Axio Imager A2 upright microscope (Zeiss, Jena, Germany) equipped with a cooled charge-coupled device (CCD) camera (Axiocam 506 color; Zeiss, Germany). Filter set 47 HE was used for aniline blue and calcofluor white. Filter set 31 was used for Congo red, PI, and FM4-64. Filter sets 09 and 38 were used for FDA. To quantify pollen tube attraction into ovules, pollen tubes were stained with 5 µM FDA dissolved in hydrated silicone oil (KF-96–100CS; Shin-Etsu Chemical, Japan). After 5 min of staining, pollen tubes were observed.
3
Cyclic Fatigue Testing of Endodontic Files
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The same instruments as in the bending test were evaluated. A self-made cyclic fatigue tester with a movable test stand (MH2-500N; IMADA, Aichi, Japan) and the X-Smart Plus endodontic motor (Dentsply Sirona) were used [23 (link)] (Figure 2 ). An artificial root canal made of stainless steel and designed with a 1.5 mm diameter, a 60° curvature, 3 mm radius of curvature, and the center of the curvature at 5 mm from the tip of the instrument was used [24 (link)]. The instruments were rotated as per the manufacturers’ recommendations (500 rpm, 1.5 N·cm in TRN; 400 rpm, 2.4 N·cm in HEDM; 300 rpm, 2.0 N·cm in PTN; and the WaveOne Gold setting in WOG), while moving the handpiece with an axial up-and-down motion of 2 mm amplitude at 300 mm/min. The canal was lubricated with silicon oil (KF-96-100CS, Shin-Etsu Chemical, Tokyo, Japan). The length of time to fracture was measured, and the number of cycles to failure (NCF) was determined as the number of revolutions (rpm) × time to fracture (seconds). All experiments were performed at room temperature.
4
Endodontic Motor Fatigue Testing
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The testing device consisted of a test stand (MH2-500N, IMADA, Aichi, Japan) with a moving stage to which the handpiece of an endodontic motor (X-Smart Plus, Dentsply Sirona) was attached. A stainless steel artificial canal with a 1.5-mm diameter, a 60° angle of curvature and a 3.0-mm radius of curvature was used 19) . The center of the curvature was located 5 mm from the tip of the instrument. The instruments (n=10, each) were rotated in the canal, while the handpiece moved with a 2-mm back-and-forth motion at 5 mm/s. Silicone oil (KF-96-100CS, Shin-Etsu Chemical, Tokyo, Japan) was used as a lubricant. The time to fracture was recorded, and the number of cycles to failure (NCF) was calculated as rpm×time to failure (min).
5
Cyclic Fatigue Evaluation of Endodontic Files
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The dynamic cyclic fatigue test [16 (link)] was conducted at RT (25 ± 1 °C) or BT (37 ± 1 °C) in an incubator made of a plastic box and a temperature controller (E5C4, OMRON Corporation, Kyoto, Japan). The testing device consisted of a test stand with a movable stage (MH2-500N, IMADA, Aichi, Japan), to which the handpiece of an endodontic motor (Tri Auto ZX2, J. Morita, Kyoto, Japan) was attached. A 17-mm-long stainless steel artificial canal with a 1.5-mm diameter, 60° angle of curvature, and 3.0-mm radius of curvature was used, and the environmental temperature of the canal was checked with a thermocouple (HOBO UX120-014 M, Onset Computer Corporation, Bourne, MA, USA). The center of the canal curvature was located 5 mm from the tip of the instrument. The instruments (n = 10, each) were fixed in the canal at 13.6 mm and rotated in combination with a 2-mm back-and-forth motion at 5 mm/s. Silicone oil (KF-96-100CS, Shin-Etsu Chemical, Tokyo, Japan) was used as lubricant. The rotational speed was set to 300 rpm for Mtwo and 500 rpm for EDM, CM, VB, RE, and JIZAI, according to the manufactures’ recommendation. The time to fracture was recorded using a stopwatch, and the number of cycles to failure (NCF) was calculated as rpm × time to failure (min).
6
Determining Instrument Fracture Resistance
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An original stainless steel three-pin device that has previously been described in detail 9, 29) was used in this study. The instrument (n=7 in each group) was fixed using three pins at an instrument curvature of 38° with a curvature radius of 5 mm, and the tip was protruded 2 mm beyond the most apical pin (Fig. 1). The experiment was performed at 300 rpm using a motor (Dentaport ZX OTR Module, J. MORITA MFG, Kyoto, Japan) until fracture occurred. Silicone oil (KF-96-100CS, ShinEtsu Chemical, Tokyo, Japan) was used to reduce friction and to minimize the release of heat during the test. A load cell (LUR-A-50NSA1, Kyowa Electronic Instruments, Tokyo, Japan) was fixed to the middle pin to determine the magnitude of the deflection load imposed by the instrument during rotation. The output of the load cell was connected to an analog-to-digital (A/D) converter with a bridge box (TUSB-S01LC, Turtle Industry, Ibaraki, Japan), and the output of the A/D converter was connected to a personal computer. The number of cycles to fracture (NCF) was recorded by measuring the time to fracture.
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