Mice were anesthetized by intraperitoneal injection of ketamine-xylazine. Blood was collected through retro-orbital bleed with a capillary tube (Fisher Scientific). Blood was allowed to clot at room temperature for 30 minutes. Serum was clarified by centrifugation and transferred to a new tube for storage at −80°C. For collection of whole blood, heparinized capillary tubes were used (Fisher Scientific).
Capillary tube
Manufactured by Thermo Fisher Scientific
Sourced in United States
Capillary tubes are narrow, cylindrical glass or plastic tubes used for various laboratory applications. They are designed to facilitate the transfer, measurement, and containment of small liquid samples or reagents through capillary action.
Automatically generated - may contain errors
Lab products found in correlation
Nucblue live readyprobe, by Thermo Fisher Scientific (2 mentions)
Fa115, by Randox (1 mentions)
Rb1007, by Randox (1 mentions)
Cd3 percp cy5, by BioLegend (1 mentions)
Cd8α alexa fluor 700, by BioLegend (1 mentions)
Cd4 apc fire750, by BioLegend (1 mentions)
Attune nxt flow cytometer, by Thermo Fisher Scientific (1 mentions)
12 protocols using capillary tube
1
Blood Collection and Serum Isolation
2
Blood Collection from Mice
3
ICG Fluorescence Detection in Tissue Phantoms
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Cuboid chicken breast tissue
phantoms of three different depths (0.5, 1.0, and 1.5 cm) were used
to assess ICG fluorescence detection through tissue. Tissue cuboids
were placed over capillary tubes [70 μL capacity, 75 mm length,
1.2 mm inner diameter (Fisher Scientific, Hampton, NH)] filled with
50 μL of 30 μM free ICG or liposomal ICG. White light
and NIR images were captured within 15 min of the preparation of capillary
tubes and tissue cuboids using a custom NIR charge-coupled device
(CCD) camera built by Hamamatsu Photonics K.K. (Hamamatsu, Japan).
Fluorescence intensity mean values of select areas, on a scale of
0 to 255 with 255 being the maximal brightness, were obtained with
the analysis function in Adobe Photoshop CS4 (Adobe Systems Inc.,
San Jose, CA).
phantoms of three different depths (0.5, 1.0, and 1.5 cm) were used
to assess ICG fluorescence detection through tissue. Tissue cuboids
were placed over capillary tubes [70 μL capacity, 75 mm length,
1.2 mm inner diameter (Fisher Scientific, Hampton, NH)] filled with
50 μL of 30 μM free ICG or liposomal ICG. White light
and NIR images were captured within 15 min of the preparation of capillary
tubes and tissue cuboids using a custom NIR charge-coupled device
(CCD) camera built by Hamamatsu Photonics K.K. (Hamamatsu, Japan).
Fluorescence intensity mean values of select areas, on a scale of
0 to 255 with 255 being the maximal brightness, were obtained with
the analysis function in Adobe Photoshop CS4 (Adobe Systems Inc.,
San Jose, CA).
4
In Vivo Tracking of DFO-NP Biodistribution
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Animals were housed in an AAALAC‐certified facility and were studied under the supervision of MGH IACUC in accordance with the approved institutional protocol (2016N000136). Before injection of DFO‐NP and DFO‐NP loaded hydrogel formulations, six‐week‐old CD‐1 mice (male; 25–30 g) from Charles River Laboratories (Wilmington, MA) were anesthetized with isoflurane and oxygen, and blood was sampled in capillary tubes (Fisher Scientific, Pittsburgh, PA) at time point 0 min by slightly cutting the end of the tail. For subcutaneous injection, DFO‐NP was dissolved in DW at a concentration of 30 wt%, and DFO‐NP/F127, DFO‐NP/HA, and DFO‐NP/HA/F127 were prepared as described above. The mice were separated into 4 groups (n = 3). 100 µL of each formulation was injected subcutaneously into the back of mice. After injection, in vivo fluorescence images were taken using the NIR imaging system (K‐FLARE) with an 800 nm channel at predetermined time points and blood samples were collected using capillary tubes at the same time. The fluorescence intensities of serum samples in capillary tubes were measured with Cytation5. 14 d after injection, mice were sacrificed to observe the biodistribution of DFO‐NP and the organs (liver, lung, spleen, kidney, intestine, and bladder).
5
Complete Blood Count Analysis Using Forcyte Analyzer
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Complete blood count was analyzed with the Forcyte analyzer hematology machine, according to the manufacturer’s instructions. Briefly, blood was collected in Eppendorf tubes containing 10 μl of 10% EDTA via retroorbital bleed using 200-μl heparinized capillary tubes (catalog no. 22-362-566, Fisher brand). Blood samples were processed within 2 hours to avoid hemolysis. The number of leukocytes (white blood cell), erythrocytes (red blood cell), lymphocytes, neutrophils, monocytes, eosinophils, and platelets was counted. Proprietary lysing agent was added to liberate hemoglobin and ultimately convert it to cyanmethemoglobin to calculate the value.
6
Hematology Analyzer for CBC Quantification
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Once EDTA anti-coagulated samples for CBC’s are received in the lab, they are immediately organized by ID number and processed on the Oxford Science hematology analyzer. The results are automatically downloaded onto an excel spreadsheet, reviewed by lab personnel, and sent to the investigator via email for further analysis. In brief, blood was collected in Eppendorf tubes containing 10 µl of 10% EDTA via retroorbital bleed using 200 µl heparinized capillary tubes (Cat# 22-362-566, Fisher brand). Blood samples were processed within 2 h to avoid hemolysis. The number of leucocytes (WBC), erythrocytes (RBC), lymphocytes (LY), neutrophil (NE), monocytes (MO), eosinophils (EO) and platelets (PLT) were counted. Proprietary lysing agent was added to liberate hemoglobin and ultimately convert it to cyanmethemoglobin to calculate the value.
7
Leatherback Turtle Nesting Blood Analysis
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From April–June, 2018, sample collection from nesting leatherbacks was conducted on Juno Beach and Jupiter Beach, Florida USA (26.836449°N, –80.041231°W to 26.943210°N, –80.071726°W). Turtles were sampled during egg deposition after the turtles had entered their nesting fixed action pattern. After disinfecting the venipuncture site (femoral rete system) with 70% isopropyl alcohol swabs, up to 10 mLs blood were collected using Vacuette® 20 G x 1½” needles (Vacuette, Greiner Bio-One, Kremsmünster, Austria) fitted into 10 mL BD sodium heparin glass Vacutainer® tubes (Becton-Dickinson and Co., Franklin Lakes, New Jersey USA). Blood was kept on ice in the field immediately after collection and was processed within 6–8 hours. PCV from whole blood was determined using capillary tubes (Fisher HealthCare, Houston, Texas, USA) centrifuged for 8 minutes at 4,200 g (5,000 rpm) using an LW Scientific C5 centrifuge (Lawrenceville, Georgia, USA) with microhematocrit tube inserts. A subset of whole blood was frozen at –80°C and the remaining blood was centrifuged at 4250 g (3,500) rpms for 10 minutes. Plasma was harvested and immediately stored at –80°C and analyzed within a maximum of 90 days.
8
Hematocrit Ratio Determination
9
Bovine Blood Sampling and Analysis
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Blood samples were collected via jugular venipuncture at 24 ± 1 h of age (after colostrum feeding) and at d 7, 14, 21, 28, 35, 42, 49, and 56 between 0900 and 1000 h. Blood was obtained using serum and sodium heparin vacutainer blood collection tubes (BD catalog #366430 and 366480, Franklin Lakes, NJ). After collection, sodium heparin tubes were placed on ice and serum tubes clotted for 20 min at room temperature. Within 1 h, all tubes were centrifuged at 3,000 × g for 20 min to harvest plasma and serum fractions. Blood hematocrit was analyzed from whole blood, and total protein was quantified from fresh plasma using a digital Brix Refractometer (MA871, Milwaukee Instruments, Rocky Mount, NC). For hematocrit, capillary tubes (Fisher Scientific #22-362-566, Pittsburgh, PA) filled with whole blood were centrifuged at 2,240 × g at 25°C for 3 min in a microhematocrit centrifuge and read using a circular hematocrit reader. Remaining serum and plasma samples were aliquoted and stored at -20°C until further analysis. Plasma concentrations of fatty acids, BHB, and glucose were determined by colorimetric, enzymatic, and GOD-PAP methods, respectively (kit #FA115, RB1007, and GL3981, Randox Laboratories Ltd., Crumlin, UK) by Randox Rx Daytona. The interassay coefficients of variation for fatty acid, BHB, and glucose analysis were 11.4, 5.7, and 8.7%, respectively.
10
CD8α+ Cell Depletion Validation
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