Myocyte volume was measured as described previously.1 (link) Isolated myocytes were placed on an inverted microscope stage (model IX51; Olympus, Tokyo, Japan). After a 5-minute stabilization, the chamber was perfused (3 mL/min). Chamber temperature was controlled by a water bath system (37°C; HAAKE; Thermo Electron Karlsruhe, Karlsruhe, Germany or 9°C; Polystat; Cole-Parmer, Vernon Hills, Ill). Cell images were displayed on a video monitor using a charge-coupled device camera (IonOptix, Westwood, Mass). Digital images of viable cells were captured using a video frame grabber (Scion, Frederick, Md) every 5 minutes (Figure 1 ). Relative cell volume change was determined as described previously.1 (link)
Polystat
Manufactured by Cole-Parmer
Sourced in United States
The Polystat is a temperature control system designed for laboratory applications. It functions as a temperature regulator, maintaining a specific temperature within a controlled environment.
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Lab products found in correlation
9 protocols using polystat
1
Measurement of Myocyte Volume Changes
2
In Vitro Left Heart Model Validation
3
Establishment of ECMO Perfusion System
4
Cardiorespiratory Monitoring in Wistar Rats
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Male Wistar rats were employed in the experiment (Charles River; aged: 20 to 24-weeks old, body weight: 300–380 g). Animal groups: vehicle (n = 7), negative control aCSF (n = 3) and TGN-073 treated group (n = 7). Rats were randomly assigned to either the treated or the vehicle group. Two rats were excluded from the study, one vehicle and one TGN-073 treated, due to sudden death and unstable blood pressure, respectively. All animals were housed in the MRI unit one week prior the study for acclimatisation to reduce variabilities in results. Food and water access was ad libitum, and 12 h dark and 12 h light cycle was maintained. Humidity (53% ± 2%), ventilation and temperature (21.5°C ± 0.5°C) were controlled automatically. The sample size was based on literature [32 (link)].
Heart rate, respiration rate and blood pressure were monitored throughout the experiments along with arterial blood gases. The rats’ mean physiological signs were: respiration 65 ± 5 breaths per minute, heartbeats per minute 400 ± 50 and mean blood pressure 95 ± 5 mmHg. In addition, the core body temperature was monitored throughout the whole experiment and was maintained at 37.0 ± 0.5°C using a rectal thermocouple and controlled by a heating pad (Polystat® Cole-Parmer). At the end of scanning, animals were euthanised.
Heart rate, respiration rate and blood pressure were monitored throughout the experiments along with arterial blood gases. The rats’ mean physiological signs were: respiration 65 ± 5 breaths per minute, heartbeats per minute 400 ± 50 and mean blood pressure 95 ± 5 mmHg. In addition, the core body temperature was monitored throughout the whole experiment and was maintained at 37.0 ± 0.5°C using a rectal thermocouple and controlled by a heating pad (Polystat® Cole-Parmer). At the end of scanning, animals were euthanised.
5
Electrochemical Synthesis of Silver Nanoparticles
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The synthesis of silver nanoparticles was conducted in a two-electrode system using an AutoLab, model PGSTAT 128N, potentiostat, controlled via software by a PC. Silver wire was used as anode and a glassy-carbon electrode was used as cathode. The electrochemical cell was filled with 100 mL of ultrapure water and the temperature was set to the desired value (preferably to 15 °C). The cell was water-jacketed. The temperature in the cell was controlled using a refrigerated circulator (Polystat, Cole Parmer, Vernon Hills, IL, USA).
The 8 cm silver electrode immersed in ultrapure water was electrochemically dissolved under chronoamperometric conditions (pulse potential and duration: 3.5 V and 3–8 h, respectively). The electrochemically active area of the silver wire was kept constant for all experiments and was equal to 5 ± 0.2 cm2. The experimental conditions (i.e., the temperature of 15 °C and the applied potential), employed for these studies resulted from our previous tests (during which those parameters were variable) and were selected as the most optimal. The exemplary chronoamperogram of metallic silver dissolution in water is presented inFigure 1 .
The 8 cm silver electrode immersed in ultrapure water was electrochemically dissolved under chronoamperometric conditions (pulse potential and duration: 3.5 V and 3–8 h, respectively). The electrochemically active area of the silver wire was kept constant for all experiments and was equal to 5 ± 0.2 cm2. The experimental conditions (i.e., the temperature of 15 °C and the applied potential), employed for these studies resulted from our previous tests (during which those parameters were variable) and were selected as the most optimal. The exemplary chronoamperogram of metallic silver dissolution in water is presented in
6
Skin Biopsy DNA Extraction Protocol
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DNA was extracted from skin biopsies using the previously described lysis protocol41 (link). Briefly, 25 mg of tissue was manually homogenized, mixed with 1 ml of 1X phosphate-buffered saline (PBS), centrifuged at 8600g for 10 min at 4 °C and the pellet was dried for 1 hr. at 50 °C. Later, 200 µl of lysis buffer containing 100 mM Tris at pH 8.5, 1 mg/ml of proteinase K and 0.05% of Tween 20 was added to the homogenate, vortexed and incubated at 60 °C in a water bath (Polystat – Cole Parmer Inc.) for 16 hrs. Post incubation, proteinase K was inactivated at 95 °C for 15 min. The lysate was cooled to room temperature and DNA was isolated by phenol chloroform extraction. DNeasy Kit (Cat No: 69504, Qiagen Inc. Netherlands) was used in samples where the tissue content was low.
7
Cysteine-Chitosan Hydrogel Fabrication
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Aqueous solutions of 4% (w/v) cysteine-chitosan and odex were prepared in distilled water. A 2% (w/v) solution of LBG in distilled water was heated at 80 °C for 30 min in a water bath to achieve complete dissolution. Equal volumes of pre-cooled solutions of cysteine-chitosan and LBG were mixed and vortexed for 1 min followed by addition of odex solution in equal volume and the polymeric solution was vortexed again for 1 min to achieve a homogeneous blend. The fabrication mixture was filled in plastic moulds and kept for cross-linking in a pre-chilled methanol-filled cryostat bath (Polystat, Cole-Parmer) at −12 °C for 48 h. After 48 h, the samples were removed and freezed at −80 °C overnight. The samples were then lyophilized (FreeZone, Labconco) and stored in air-tight vials at 4 °C until further use.
8
Viscosity of Aqueous HPβCD Solutions
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The viscosity measurements of aqueous HPβCD solutions were performed with a Brookfield model DV-I + (USA) viscometer equipped with a Polystat circulating thermostatic water bath (Cole Palmer, Vernon Hills, IL, USA) set to 25 °C. All samples were analyzed in triplicate in order to calculate average values and SD.
9
Ultrasonic Sonication of SPI Solutions
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Solutions of SPI were sonicated for 5, 10, 15 and 20 min using an ultrasonic processor Vibra Cell Sonics, model VCX 750 (maximum net power output: 750 W) at a frequency of 20 kHz, 4.27 ± 0.71 W and an amplitude of 20% without any booster. A 13 mm high grade titanium alloy probe threaded to a 3 mm tapered microtip was used to sonicate 10 ml of solution at 6% wt/wt. Samples contained into glass test tubes were, in turn, immersed into a glycerine-jacketed circulating constant temperature cooling bath at 0.5 °C to dissipate most of the heat produced during sonication and 75, 80 and 85 °C for temperature combined treatments (Polystat, Cole-Parmer).
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