Adult I. scapularis were acquired from BEI Resources (American Type Culture Collection) and the National Tick Research and Education Resource (Oklahoma State University). Groups of 10–12 ticks were housed in 20 ml glass scintillation vials (VWR International, PA, USA) fitted with strips of paper towel as refuge and substrate for climbing, and with a mesh-covered hole (approx. 1 cm) in the lid to enable air exchange. As ticks are prone to desiccation, vials were kept at high relative humidity (85–95%) in a vessel (d = 26 cm, h = 30 cm) containing a saturated solution of K2SO4 (99% purity; Alfa Aesar, ON, Canada). To minimize the risk of tick escape, the vessel was retained in a plexiglass box (50 × 35 × 35 cm) which was kept at 22°C and a 14 : 10 light/dark cycle. To prevent mould/fungal growth, vials were washed weekly with Sparkleen (Thermo Fisher Scientific, MA, USA) and dried at 100°C for more than 1 h. Monthly, the vessel was washed and sterilized with Sparkleen and 70% ethanol, respectively, and the K2SO4 solution was replaced.
Sparkleen
Manufactured by Thermo Fisher Scientific
Sourced in United States
Sparkleen is a laboratory cleaning solution designed to effectively remove a wide range of contaminants from lab equipment and glassware. The product's core function is to provide a thorough and efficient cleaning process to maintain the integrity and performance of laboratory instruments and materials.
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Lab products found in correlation
10 protocols using sparkleen
1
Tick Rearing and Preservation Protocol
2
Profiling Microbial Volatile Organic Compounds
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Headspace volatiles were collected from all strains of microbes that elicited significant behavioural responses by ticks in Y-tube olfactometer experiments. To this end, 12 MHA plates were inoculated with a microbe of interest and incubated for approximately 20 h. These plates, with open lids, were then placed into an aeration chamber (diameter = 19 cm: height = 29.5 cm) connected to a vacuum pump (Neptune Dyna-pump). Charcoal-filtered air was drawn at a flow rate of 1 l·min−1 for 20 h through the chamber and through a glass column (6 mm outer diameter × 150 mm) containing 200 mg of manufacturer-preconditioned Porapak-Q adsorbent (50–80 mesh; Waters Associates, Milford, MA, USA), or 200 mg of the adsorbents Carbosieve SIII (60/80 mesh; Supelco, PA, USA) and Tenax TA (35/60 mesh, Chromatographic Specialties, Brockville, ON, CA) (2.7 : 1). Volatiles were desorbed with one rinse of ether (2 ml), which then contained 240 plate-hour-equivalents (240 PHEs; 12 microbe-inoculated ager plates × 20 h of volatile captures) of headspace volatile extract (HVE). Volatile extracts were concentrated to 0.5 ml under a stream of nitrogen, and were kept at 4°C prior to analyses. All glassware was cleaned with Sparkleen (Thermo Fisher Scientific, MA, USA), rinsed with distilled water, and oven-dried at 130°C prior to starting a new aeration.
3
Fluorescent Polymer Coatings for Photoluminescence Analysis
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TiN and Au–TiN samples were coated with a fluorescent thin-film before PL measurements. A fluorescent conjugated polymer poly(9,9-dioctylfluorene-alt-benzothiadiazole) (F8BT) was dissolved in chloroform to a concentration of 4.2 mg mL−1 (6.3 mg of F8BT was dissolved in 1.5 mL of chloroform). The solution was heated in a water bath at 45 °C and simultaneously stirred at 5000 rpm for 15 min and then sonicated for 15 min. Finally, 50 μL of the F8BT solution was dynamically spin coated onto the samples at a spin speed of 5000 rpm for 60 seconds. The same spin coating conditions were also used to coat a cleaned glass substrate with F8BT. Prior to spin coating, the glass substrate was cleaned in an ultrasonic bath containing detergent (0.5 wt%; Sparkleen, Fischer Scientific) for 10 min., and, subsequently, in a 50 : 50 solution of hydrochloric acid and ethanol for 10 min. The substrate was triple rinsed in ultrapure water (Mili-Q) after each bath cleaning step. The bright-field/dark-field imaging was performed on an inverted microscope (Axio Vert.A1, Carl Zeiss Microscopy, LLC.) coupled to an imaging spectrometer (Shamrock SR303i-A, Andor Technology Ltd.). Photoluminescence imaging and spectroscopy were carried out using an excitation source (X-Cite® 120Q, Excelitas Technologies Corp.) combined with a 365 excitation filter and a 397 nm long pass filter.
4
Microfluidic Device Fabrication via Stereolithography and Soft-Lithography
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A combination of stereolithography and soft-lithography techniques were used to generate MesoFind microfluidic devices. Briefly, 3D printed positive molds were fabricated using stereolithography (μMicrofluidics Edition 3D Printer from Creative CADworks, Canada). Polydimethylsiloxane (PDMS) (Dow Corning, USA) was cast onto the positive molds and baked for 2 hours at 70°C to generate negative molds. The negative molds were treated in a saturated detergent solution (Sparkleen, Thermo Fisher Scientific, USA) in 70% ethanol for one hour at room temperature. PDMS was then cast onto the negative molds and again baked for 2 hours at 70°C to generate PDMS positive imprints. The PDMS imprints were peeled and bonded onto thickness #1 glass slides (Ted Pella, USA) using plasma treatment and were then incubated overnight at 100°C. The inlets and outlets were punched and high-purity silicone tubing (McMaster-Carr, USA) was inserted. Tubing connections were sealed with liquid PDMS followed by an hour of baking at 70°C. Chips were flushed with 0.1% Pluronic F-68 (P5556, Sigma-Aldrich, USA) in phosphate buffer saline (PBS) (Wisent Bioproducts, Canada) overnight before usage.
5
Microfluidic Device Fabrication for Cell Capture
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Positive molds were fabricated by a stereolithographic 3D printer (μMicrofluidics Edition 3D Printer, Creative CADworks, Canada) using the “CCW master mold for PDMS” resin (Resinworks 3D, Canada). The layer thickness is set to 50 μm. Negative molds were fabricated by casting PDMS (Dow Chemical, USA) on positive molds and baked at 70°C for 2 hours. Negative molds were then treated by saturated detergent solution (Sparkleen, Thermo Fisher Scientific, USA) in 70% ethanol at room temperature (RT) for at least an hour. PDMS-positive replicas were generated by casting PDMS on negative molds and baked at 70°C for 2 hours. The cured replicas were then peeled off, punched, and plasma bonded to thickness no. 1 glass coverslips (Ted Pella, USA). The bonded chips were left in a 100°C oven for 30 min to secure a robust bonding. Afterward, the silicon tubing was attached to the inlet and outlet of the device. Before use, the devices were conditioned with 1% Pluronic F68 (Sigma-Aldrich, USA) in phosphate-buffered saline (PBS) for at least 1 hour to reduce the nonspecific adsorption. Each device was sandwiched between two arrays of N52 NdFeB magnets (K&J Magnetics, USA; 1.5 mm by 8 mm) with alternating polarity. A syringe pump (Chemyx, USA) was used for the duration of the cell capture process.
6
Coverslip Surface Functionalization Methods
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Cover glass (18 mm, 0.12–0.17 mm thickness, Matsunami micro cover glass) were first cleaned with 10% detergent (Sparkleen, Fisher Scientific, USA) and sonicated (Aristocrat Ultrasonic, Healthco, USA) for 10 min, followed by three washes in distilled water and allowed to dry vertically in a holder. Following this, some covers slips were soaked in 95% Ethanol (Sigma-Aldrich, USA) for 10 min, while another set was soaked in 2% n-octadecyl trichlorosilane (ODS) for 10 min. Finally, another set of coverslips was subjected to RFGDT, as described below. Except for the RFGDT, all samples were sterilized under UV light or 15 min before cell seeding.
7
Fabrication of PDMS Microfluidic Devices
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A poly(dimethylsiloxane) (PDMS) Sylgard® 184 elastomer kit was purchased from Ellsworth Adhesives. The pre-polymer and curing agent were well-mixed at a 10:1 ratio. The mixture was degassed in a vacuum chamber for 1 h and poured on a clean mold with the desired channel, patterned with SU-8 photoresist. The mold was then degassed for an additional 1 h before being placed in an 80°C oven for 30 min. The PDMS was carefully removed from the mold, and excess PDMS was removed using a razor blade. The channel was cleaned with Sparkleen (Fisherbrand), rinsed with deionized water, sprayed with 70% ethanol and rinsed again with deionized water. The PDMS was dried with filtered, in-house air before covering with scotch tape until ready for plasma cleaning. Cover slips were cleaned and dried in a similar manner before sealing the microdevices. To seal microdevices, the microchannel and cover slip were plasma cleaned for 1 min, sealed and placed in the oven for 30 min. The resulting device channel dimensions were 100 μm in height with a length and width of 4 mm and 2 mm, respectively.
8
Aerosol Exposure System Setup and Maintenance
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The aerosol generator assembly (Biaera Technologies, LLC, USA) in the Duke RBL was used for all aerosol experiments. A detailed diagram of aerosol chamber exposure system that includes components such as biosampler, nebulizer, chamber pressure sensor, relative humidity, temperature controller, sampling port and connection of the port to animal exposure chamber etc., and their description has been previously published [14 (link)]. The Duke RBL is a fully commissioned BSL3/ABSL3 facility housed in the Global Health Research Building (GHRB), Duke Human Vaccine Institute (DHVI), Duke University School of Medicine, Durham, NC. Standard operating procedures were followed to maintain sterile conditions and whole-body aerosol exposures carried out as previously described [14 (link), 15 (link)]. The biosampler, nebulizer, 6-jet nozzles, and O-rings were routinely examined for any deposition or damage such as fraying or decay. These components were properly cleaned using Sparkleen (Cat#04-320-4, Fisher Scientific) and warm water in a Precision Needle-Tip Squeeze Bottle (Cat#1902T61, McMaster-Carr) followed by thorough rinsing or were replaced when required. All components were routinely maintained as per manufacturer’s instructions (Biaera Technologies).
9
Mosquito Grooming Behavior and Particulate Formulations
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Example 10
This example demonstrates that female mosquitoes are able to remove particulate formulations over time due to their grooming behavior. Forty gravid females were introduced into an autodissemination station charged with powder formulation, the entrances blocked, and the station placed into a cubic 2.2 m screened cage provisioned with sucrose solution. The station portals were reopened after 30 min, permitting the now powder-contaminated mosquitoes to quickly exit the station and enter the cage. The station was then removed. Five gravid females were individually captured with an aspirator after 0, 1, 2, 6, 12, 24, and 48 hr, transferred into centrifuge tubes, and killed by freezing at −4° C. for 4 hr. The mosquitoes were placed into individual 200 μĩ drops of 0.1% detergent (Sparkleen®, Fisher Scientific, Pittsburgh, Pa. USA) to remove adhering powder, and the retrieved particles counted. The experiment was replicated three times with environmental conditions of 26-28° C., 60-75% RH, and 16L:8D photoperiod.
10
Fly Behavioral Response to Stimuli
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The response of flies to visual and olfactory stimuli was tested in 2-choice experiments 1–11 (Fig 1 ) using BioQuip® (Compton, CA) wire mesh cages (61 × 61 × 61 cm) with a plated grey base (BioQuip®, Compton, CA, USA). Each cage was illuminated from above with fluorescent lights (Phillips F32TA, Amsterdam, The Netherlands; light intensity in cage: 236 Lux). For each experimental replicate, 100 recently eclosed (1- to 3-day-old) cold-sedated flies of mixed sex (approximate sex ratio 50:50) were introduced into a cage, allowing them to acclimate for 2 h prior to the start of the experiment. The stimuli was tested as part of an inverted “bottle trap” consisting of a 500-mL plastic soda bottle with the inverted top providing a cone-shaped funnel (11.5 cm long × 0.6 cm bottom diameter × 7.9 cm top diameter; Fig 2A ) that rested on the bottom half (20 cm long × 7 cm diameter; Fig 2A ). The funnel was covered with construction paper (11.5 cm long × 0.6 cm diameter) of a specified color as the visual test cue (Fig 2D ) and wrapped the bottom half with green construction paper (Fig 2A ), randomly assigning test stimuli to opposite corners of the bioassay cage. A mote of Sparkleen™ (Fisher Scientific Co. Pittsburgh, PA, USA) and water (0.5:5) in the trap bottom drowned the flies that entered the trap. Trap captures were scored after a 6-h experimental duration.
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