L4-stage hermaphrodites were placed on a seeded NGM plate and incubated overnight at either 20 °C. After incubation, two to seven young adult hermaphrodites were transferred to a drop of M9 buffer containing levamisole (0.5 mM) on a 10% agarose pad (Nacalai Tesque), which was created inside an annular silicone sheet placed on a glass slide. The transferred worms were enclosed and immobilized on a coverslip. Time-lapse imaging was performed using a CSU-W1 spinning-disc confocal system (Yokogawa Electric Corporation) attached to an inverted microscope IX71 (Olympus) and an sCMOS camera Sona (Andor). The glass slide containing the specimen was held on a constant-temperature unit ThermoPlate (Tokai Hit) and maintained at 20 °C. The exposure settings and interval for photography were 500 ms with 20% laser power at 470 nm excitation, 500 ms with 80% laser power at 555 nm excitation, and 150 ms with DIC. Visualization was performed with a software Micro-Manager v2.0.0. Movies were created using ImageJ (version: 1.53k) from the sequential images obtained.
Thermo plate
Manufactured by Tokai Hit
Sourced in Japan
The Thermo Plate is a laboratory equipment used for temperature control. It provides a stable and consistent temperature environment for various applications.
Automatically generated - may contain errors
Lab products found in correlation
Ketamine hydrochloride, by Merck Group (3 mentions)
Axio observer z1, by Zeiss (3 mentions)
Zen system, by Zeiss (3 mentions)
Colibri 2 led light, by Zeiss (3 mentions)
Lsm 880, by Zeiss (2 mentions)
Illuminator hxp 200c, by Zeiss (2 mentions)
Plan apochromat objective, by Zeiss (2 mentions)
Axio zoom v16, by Zeiss (2 mentions)
Ketamine, by Merck Group (2 mentions)
Xylazine, by Merck Group (2 mentions)
Axiolab, by Zeiss (1 mentions)
Observer z1, by Zeiss (1 mentions)
Im 300 microinjector, by Narishige (1 mentions)
Eclipse fn1, by Nikon (1 mentions)
A1r confocal laser scanning microscopy system, by Nikon (1 mentions)
Fastcam viewer software, by Photron (1 mentions)
Fv1000, by Olympus (1 mentions)
Infinite f500, by Tecan (1 mentions)
Microdroplet culture dishes, by Vitrolife (1 mentions)
Polyvinylpyrrolidone, by Irvine Scientific (1 mentions)
32 protocols using thermo plate
1
Time-lapse Imaging of C. elegans Development
2
Evaluation of Fertile Dog Semen
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Semen from 3 fertile dogs was collected and frozen as described below. In all cases, dogs were referred to Department of Veterinary Medical Sciences of Alma Mater Studiorum -University of Bologna for sperm evaluation and informed consent of the owner was taken. An informed consent of the owner of bitches subjected to ovariectomy/ovariohysterectomy of was also taken.
The three fractions of the ejaculates were collected in calibrated plastic vials by digital manipulation. The sperm-rich fraction was evaluated before freezing for volume, concentration, and rate of morphologically normal spermatozoa. The volume was measured by a calibrated micropipette, and sperm concentration was determined with a Bürker chamber, after dilution of the sperm suspension 1:40 with 10% formol buffered saline to immobilize spermatozoa and counted using a phase contrast microscope (400x; Axiolab; Zeiss, Italy) equipped with a warming plate (37°C; Thermo Plate; Tokai Hit, Japan) as reported in [14] (link). Percentages of morphologically normal spermatozoa were determined at the same microscope (1000x) after dilution of semen 1:1 with 10% formol buffered saline, and at least 200 spermatozoa per sample were examined.
For the study only samples with total motility (TM) ≥ 90%, rate of morphologically normal spermatozoa ≥ 90% were used.
The three fractions of the ejaculates were collected in calibrated plastic vials by digital manipulation. The sperm-rich fraction was evaluated before freezing for volume, concentration, and rate of morphologically normal spermatozoa. The volume was measured by a calibrated micropipette, and sperm concentration was determined with a Bürker chamber, after dilution of the sperm suspension 1:40 with 10% formol buffered saline to immobilize spermatozoa and counted using a phase contrast microscope (400x; Axiolab; Zeiss, Italy) equipped with a warming plate (37°C; Thermo Plate; Tokai Hit, Japan) as reported in [14] (link). Percentages of morphologically normal spermatozoa were determined at the same microscope (1000x) after dilution of semen 1:1 with 10% formol buffered saline, and at least 200 spermatozoa per sample were examined.
For the study only samples with total motility (TM) ≥ 90%, rate of morphologically normal spermatozoa ≥ 90% were used.
3
Microinjection Technique for Cell Transplantation
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Glass thin-capillary tubes (GD-1 with filaments; Narishige Co., Tokyo, Japan) pulled on a P-97/IVF micropipette puller (Sutter Instrument Co., Novato, CA, USA) were used for transplant. After pulling, the capillary was bent at a 30° angle at 1.0–1.2 mm from the tip by a MF-1 micro-forge (Technical Products International, St. Louis, MO, USA). The injection capillary and a holding pipette were connected to an IM 300 Microinjector (Narishige) and hydraulic micromanipulator (NT-8, Narishige), respectively. The tip of the capillary and cells were visible on an inverted microscope (Observer Z1, Zeiss Co., Germany). The stage was kept at room temperature during microinjection. After injection, the stage was exchanged for a mechanical stage (ThermoPlate, MATS-555AXK and MATS-555RN, Tokai Hit Co., Shizuoka, Japan) to incubate cells at 37 °C. The whole injection device was placed on a vibration isolator system (ST-X, Showa Science Co., Tokyo, Japan).
4
Sensor-Integrated Microfluidic Platform
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The microfluidic set-up comprised the sensor-integrated microfluidic platforms mounted on top of a heated glass plate (ThermoPlate® Tokai-hit, Japan) within an inverted microscope (IX70, Olympus), connected (Marprene Manifold Tubes 0.25 mm, Watson-Marlow and Tubing FEP Nat 1/16 × 0.030, Idex Health and Science) to a peristaltic pump (205S/ CA manual control variable speed pump, Watson Marlow) as well as 16 optical fibers (length 1 m, outer diameter 2.2 mm, fiber diameter 1 mm, Pyroscience) via four FireSting optical oxygen meters (2x FireStingO2, 2x FSPRO-4, Pyroscience, Germany).
The cell culture medium and the nanoparticle solutions were preconditioned employing a desktop incubator, connected to the peristaltic pump using FEP tubing (Tubing FEP Nat 1/16 × 0.030, Idex Health and Science). Both the oxygen meters and the microscope were connected to a workstation enabling the interaction through the respective interfaces (Cellsens, Olympus and OxygenLogger, Pyroscience). Fig. 3 comprises a schematic representation of the microfluidic set-up.
The cell culture medium and the nanoparticle solutions were preconditioned employing a desktop incubator, connected to the peristaltic pump using FEP tubing (Tubing FEP Nat 1/16 × 0.030, Idex Health and Science). Both the oxygen meters and the microscope were connected to a workstation enabling the interaction through the respective interfaces (Cellsens, Olympus and OxygenLogger, Pyroscience). Fig. 3 comprises a schematic representation of the microfluidic set-up.
5
Visualizing Micelle Crossing the Blood-Brain Barrier
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The mice were anaesthetized with 2.5% isoflurane (Abbott, North Chicago, IL) using a Univentor 400 anaesthesia unit (Univentor; Zetjun, Malta). The skulls of mice were partially cut open around the ROIs without damaging blood vessels. The mice were directly placed on a thermoplate (Tokai Hit, Tokyo, Japan), and a cover slip (Muto Pure Chemicals, Tokyo, Japan) was attached with proper pressure to flatten the brain surface. Images of the micelle crossing the BBB were acquired with an A1R confocal laser-scanning microscopy system (Nikon Corp., Tokyo, Japan) attached to an upright ECLIPSE FN1 (Nikon Corp.).
6
Cryopreservation and Viability of Fibroblast Sheets
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Fibroblast sheets were soaked in 300 μL of cell preservation solution per well in 24-well plates and 1000 μL of cell preservation solutions per well in 12-well plates. For freezing using the 3D Freezer, multi-well plates were placed in a precooled 3D Freezer at −35 °C for 20 min. Multi-well plates were stored in a −80 °C freezer for 2 h. For freezing using the Program Freezer, the parameters for cooling were as follows: 4 °C for a 5-min hold, −2 °C per min down to −30 °C, −30 °C for a 5-min hold, and −1 °C per min down to −80 °C. Multi-well plates were stored in a −80 °C freezer for 2 h. For thawing the cells, the multi-well plates were placed on a Thermo plate (Tokai Hit., Co., Ltd., Shizuoka-ken, Japan) at 37 °C for 14 min. Fibroblast sheets were soaked in phosphate-buffered saline (PBS) (Cell Science & Technology Institute) twice for washing. To measure cell viability, fibroblast sheets were incubated in a 24-well plate (1 mL medium/well) with equal amounts of CTS™ AIM V™ SFM and HFDM-1 (+) medium supplemented with 2% NeoSERA at 37 °C in a humidified 5% CO2 incubator for 3 days. The supernatant was collected for enzyme-linked immunosorbent assay (ELISA). Non-frozen fibroblast sheets were soaked in PBS and stored at room temperature instead of freezing and storing at −80 °C.
7
Microfluidic Rheology of Blood Cells
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The experimental set-up included a syringe pump (KDS101 CE, KD Scientific, Holliston, MA, USA) to control the inlet flow rates, as well as an inverted microscope (Olympus IX71 from Olympus, London, UK) equipped with a thermal plate (ThermoPlate from Tokai-Hit, Fujinomiya, Japan) to place and heat the PDMS device, and a thermometer to measure the PDMS temperature (ThermoWorks, Inc., Salt Lake City, UT, USA) (Figure 2 ).
Images and videos were obtained using a Photonics camera (Photron 1024 PCI FASTCAM Camera from Photron, West Wycombe, UK) and recorded at a frame rate between 1000 and 3000 frames per second (fps) and a shutter speed of 1/10,000 s. The CFA distance was measured using the Photron FASTCAM Viewer software.
Blood samples were from anonymous donors and stored at 4 °C for no more than three days after extraction. The whole blood samples were centrifuged three times at 2500 rpm for 4 min to remove the plasma and wash the cells in PBS.
Cells were re-suspended in their autologous plasma, or in PBS, at a concentration of 20%, to study the effect of cell flow suspended in these two solutions at a wide range of temperatures and shear rates.
Images and videos were obtained using a Photonics camera (Photron 1024 PCI FASTCAM Camera from Photron, West Wycombe, UK) and recorded at a frame rate between 1000 and 3000 frames per second (fps) and a shutter speed of 1/10,000 s. The CFA distance was measured using the Photron FASTCAM Viewer software.
Blood samples were from anonymous donors and stored at 4 °C for no more than three days after extraction. The whole blood samples were centrifuged three times at 2500 rpm for 4 min to remove the plasma and wash the cells in PBS.
Cells were re-suspended in their autologous plasma, or in PBS, at a concentration of 20%, to study the effect of cell flow suspended in these two solutions at a wide range of temperatures and shear rates.
8
Characterization of Liposomal Nanocomposites
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Dynamic light scattering (DLS) measurements were performed using a Zetasizer Nano series (Malvern Inst. Ltd., Malvern, UK). A confocal laser scanning microscope (FV-1000, Olympus, Tokyo, Japan) was used to take the liposome image using a stage thermocontrol system (Thermoplate, Tokai Hit, Shizuoka, Japan). Fluorescence spectra and UV-vis absorption was taken by using a microplate reader (Infinite F500; TECAN, Ltd., Männedorf, Switzerland). Transmission electron microscopy (TEM) images of QDs, Fe3O4, liposomes, and their nanocomposites were obtained by JEOL TEM (JEOL, Tokyo, Japan). Electrochemical DPV was performed by an SP-150 (BioLogic.inc, Tokyo, Japan) in a saturated Ag/AgCl, with a conventional three-electrode cell consisting of a glassy carbon disk electrode (4 mm in diameter) as the counter, reference, and working electrodes, respectively (EC frontier, Tokyo, Japan).
9
Modulating Schistosome Motility with Transient Receptor Potential Mucolipin (TRPML) Compounds
10
ICSI Blastocyst Culture and Transfer
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All eggs underwent ICSI immediately after PLM (re-)assessment. They were placed into 5-μl droplets of a prewarmed HEPES/MOPS-buffered medium, covered by mineral oil on plastic microinjection dishes (Nunc™ IVF Petri Dishes, #150270, Thermo Fisher Scientific, Waltham, USA). ICSI was routinely performed according to the standard protocol using ICSI/holding micropipettes (#002-5-30/#001-120-30, Microtech IVF, The Czech Republic), polyvinylpyrrolidone (#90121, Irvine Scientific, USA), and Eppendorf (Hamburg, Germany) micromanipulation system equipped with thermoplate (TokaiHit, Japan). After ICSI, injected oocytes were placed into 30-μl droplets of CO2-dependent CSC medium covered with mineral oil in micro-droplet culture dishes (#16003, Vitrolife, Sweden) and individually cultured at 37 °C in a humidified atmosphere of 5% O2 and 6% CO2. Embryo development was followed until day 5 or day 6 when good-looking blastocysts were either transferred in a fresh cycle or cryopreserved (Rapid-i, #10119, Vitrolife, Sweden). Blastocysts were chosen for transfer based on morphological criteria by clinical embryologists who were blinded to the PLM assessment results.
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