During the springs of 2011, 2013, and 2014 health assessments were performed in populations of bog turtles selected by biologists as representative of healthy populations as well as populations of concern due to recent mortality events. Most turtles were located in fens by walking the areas and gently probing the soft substrate or by direct visualization of animals. Turtles then were manually restrained and the choana and then cloaca were swabbed with a single rayon-tipped, plastic shaft, fine tip swab (Medical Wire and Equipment, Wiltshire, England). Samples were collected from the states of Massachusetts (2 sites; April and May, 2011), New York (2 sites; May, 2011), New Jersey (10 sites; May, 2013 and 2014), Pennsylvania (3 sites; May, 2013), and Delaware (2 sites; May, 2013 and 2014). Swabs were placed in 2ml polypropylene micro tubes with screw caps (Sarstedt, Nümbrecht, Germany), then stored at 4° C for up to 48 hours and at -80° C for long term storage. The total sample population (n = 230) included bog (n = 204), spotted (n = 17) and wood (n = 9) turtles (Table 1 ).
Screw cap
Manufactured by Sarstedt
Sourced in Germany
Screw caps are used to securely close various laboratory containers and vessels, providing a tight seal to prevent leakage or contamination. They are made of durable materials and designed to be easily screwed on and off as needed.
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Lab products found in correlation
6 protocols using screw cap
1
Bog Turtle Health Assessments
2
Biobanking of Alzheimer's CSF Samples
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Anonymous CSF samples that had inadequate clinical information or were not fit for clinical validation were collected from Alzheimer Center Biobank at the VU University Medical Center (VUmc, Amsterdam, The Netherlands) and University Medical Center (Göttingen, Germany). The samples were pooled and aliquoted as 500 μl aliquots. Samples collected from the Biobank at VUmc were used in α-syn stability and adsorption experiments while those from University Medical Center Göttingen were used in investigations done on adsorption of α-syn. Aliquots were stored at −80°C in 1.5 ml polypropylene tubes with screw caps (Sarstedt, Nümbrecht, Germany). All samples were blinded, and donors gave written informed consent at study entry for the use of clinical information and CSF material for scientific research purposes. The study was conducted according to the revised Declaration of Helsinki and Good Clinical Practice guidelines and approved by the local ethics committee of the VU University Medical Center.
3
Plasma and Buffy Coat Collection for Trypanosoma evansi Detection
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The blood collected in the heparinised Venosafe tubes was centrifuged for 10 min at 1,000 rpm and plasma was collected with a single use plastic transfer pipette into 2 mL tubes with screwcaps (Sarstedt, Nümbrecht, Germany). Plasma was stored at 4°C until testing for specific antibodies with CATT/T.evansi and subsequently frozen at −20°C. From the remaining blood specimen, 500 μL of buffy coat layer were collected by means of a micropipette with a filter tip and mixed with an equal volume of guanidium EDTA buffer (GEB; 6 M guanidium chloride, 0.2 M EDTA, pH 8.0) and stored at ambient temperature until DNA extraction [68 (link)]. Of those animals that were parasitologically positive, part of the buffy coat was collected for cryopreservation in liquid nitrogen for later isolation of the parasite according to Pyana et al. [69 (link)].
4
Androstenedione Extraction from Biological Fluids
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The internal standard solution (10 µL) was added to aliquots of 100 µL of serum/plasma/calibrators/controls and incubated on an overhead shaker for 10 min at room temperature; 0.5 mL polypropylene microtubes were used together with screw caps (Sarstedt, Nümbrecht, Germany). Subsequently, the samples were treated with 200 µL of the precipitating agent (ZnSO4 in water (89 g/L)/methanol 1/4 v/v), mixed on a thermomixer for 10 min at 1400 rpm (5382 thermomixer C, Eppendorf AG, Hamburg, Germany), and centrifuged for 10 min at 16,000 rcf and 10 °C (centrifuge 5430 R, Eppendorf). For the final sample purification, 200 µL aliquots of the supernatant were loaded onto Oasis prime HLB (30 mg) cartridges from Waters (Eschborn, Germany) by centrifuging the samples for 2 min at 50 rcf (centrifuge 5810 R, Eppendorf). The cartridges were washed with 500 µL of water (centrifuging for 3 min at 100 rcf), after which the purified androstenedione was eluted by applying two times the volume of 100 µL acetonitrile (centrifuging for 3 min at 50 rcf). Both phases were collected in a HPLC vial, and the samples were analyzed in duplicate. A schematic overview is given in Supplemental Fig. 1 .
5
Isolation and Characterization of MRSA Biofilms
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For the present study, we used an established MRSA strain (ATCC 33591). One hundred and seventy-four clinical samples of MRSA were isolated in Fukuoka University Hospital, one of which (OJ-1) was from an ulcerated wound [13 (link)] and four (T12, T34, T41 and T144) were from blood [14 (link)]. These particular bacterial isolates were selected because of their superior ability to form stable biofilms. MRSA samples were stored in a deep-freeze, and upon thawing were incubated on tryptic soy agar (TSA) (Becton Dickinson) containing 0.5 % NaCl. Upon colony formation, one colony was inoculated in 5 ml tryptic soy broth (TSB) (Becton Dickinson) in a 12 ml plastic test tube with a screw cap (Sarstedt) at 37 °C. Cultures that achieved stable growth were subsequently cultured on agar, and the colonies formed were stored at 4 °C and used for experiments within 1 month.
6
Ex Vivo Blood Irradiation Simulation
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This simulation reproduces the irradiation geometry of a blood sample in an 8 ml vial with a screw cap (Sarstedt, No. 60.542.007, Germany). The vial dimensions are detailed in Table 1 .
The vial material was set as polypropylene with a density of 0.9 g/cm3 and an elemental composition of hydrogen and carbon (mass fractions: 0.143711 and 0.856289, respectively) [17] . Inside of the vial, a cylindrical volume composed of blood (7ml) plus water (1ml) (radioactive solution) is located. The dimensions of the cylindrical volume were 8 ml (total volume), 7.2 mm (radius), and 49.12 mm (height).
The vial was located in a simulation space of 0.5 m × 0.5 m × 0.5 m without surrounding material (vacuum). Inside this space, we introduced a second space of 0.1 m × 0.2 m 0.1 m filled with air. These conditions reproduce the ex vivo irradiation conditions in the laboratory. In addition, they delimit the simulation space to the vial and the immediate surroundings.
The vial material was set as polypropylene with a density of 0.9 g/cm3 and an elemental composition of hydrogen and carbon (mass fractions: 0.143711 and 0.856289, respectively) [17] . Inside of the vial, a cylindrical volume composed of blood (7ml) plus water (1ml) (radioactive solution) is located. The dimensions of the cylindrical volume were 8 ml (total volume), 7.2 mm (radius), and 49.12 mm (height).
The vial was located in a simulation space of 0.5 m × 0.5 m × 0.5 m without surrounding material (vacuum). Inside this space, we introduced a second space of 0.1 m × 0.2 m 0.1 m filled with air. These conditions reproduce the ex vivo irradiation conditions in the laboratory. In addition, they delimit the simulation space to the vial and the immediate surroundings.
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