BALF was collected by endotracheal intubation with 300 μl ice-cold PBS twice and then centrifuged at 500 g for 10 min at 4 °C. The supernatants were used for further ELISA. The total cells were resuspended with 50 μl PBS and counted via the Mindray BC-5000Vet automated hematology analyzer (Mindray, Shenzhen, CHN). (See Fig. 6 ).
Bc 5000vet
Manufactured by Mindray
Sourced in China
The BC-5000Vet is a hematology analyzer designed for veterinary use. It provides automated analysis of complete blood count (CBC) parameters in animal samples. The instrument performs red blood cell, white blood cell, and platelet counts, as well as hemoglobin measurement and other key hematological tests.
Automatically generated - may contain errors
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59 protocols using bc 5000vet
1
BALF Collection and Cell Quantification
2
Automated Hematology and Biochemistry Analyses
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The automated analyzer was used for measurements of CBC (Mindray BC-5000VET, Shenzhen
Mindray Bio-Medical Electronics Co., Ltd., Shenzhen, China) and serum chemistry profiles
(BUN, Cr, ALT, TP and albumin) (ILAB 650 Chemistry Analyzer, Diamond diagnostics,
Holliston, MA, USA). The concentration of serum SDMA was measured by commercial analyzer
(Vcheck V200, Bionote Co., Ltd., Gyeonggi-do, South Korea). The information from
manufacturer showed the close relationship between concentration of SDMA obtained from
this kit and from gold standard “I” laboratory (R2=0.9908). This method had the
lowest detecting value of 10 µg/dl and highest detecting value of 100 µg/dl. For iron
parameters, serum iron concentration and unsaturated iron binding capacity (UIBC) were
analyzed by the standard colorimetric Ferrozine method (Cobas c501, Roche Diagnostics,
Indianapolis, IN, USA). The TIBC was calculated as TIBC=UIBC + serum iron concentration.
Serum hepcidin levels were measured by Sandwich quantitative ELISA technique using canine
hepcidin (HEPC) ELISA kit (Catalogue # MBS010437; MyBiosource Inc., San Diego, CA, USA).
The percent saturation (%Sat) was calculated as the ratio of serum iron concentration to
TIBC.
Mindray Bio-Medical Electronics Co., Ltd., Shenzhen, China) and serum chemistry profiles
(BUN, Cr, ALT, TP and albumin) (ILAB 650 Chemistry Analyzer, Diamond diagnostics,
Holliston, MA, USA). The concentration of serum SDMA was measured by commercial analyzer
(Vcheck V200, Bionote Co., Ltd., Gyeonggi-do, South Korea). The information from
manufacturer showed the close relationship between concentration of SDMA obtained from
this kit and from gold standard “I” laboratory (R2=0.9908). This method had the
lowest detecting value of 10 µg/dl and highest detecting value of 100 µg/dl. For iron
parameters, serum iron concentration and unsaturated iron binding capacity (UIBC) were
analyzed by the standard colorimetric Ferrozine method (Cobas c501, Roche Diagnostics,
Indianapolis, IN, USA). The TIBC was calculated as TIBC=UIBC + serum iron concentration.
Serum hepcidin levels were measured by Sandwich quantitative ELISA technique using canine
hepcidin (HEPC) ELISA kit (Catalogue # MBS010437; MyBiosource Inc., San Diego, CA, USA).
The percent saturation (%Sat) was calculated as the ratio of serum iron concentration to
TIBC.
3
Hematology and Clinical Chemistry
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Blood analyses were done with 30 μl of blood in Hemocytometer Mindray BC-5000Vet. Kidney and Liver parameters were measured with 100 μl of blood in MNCHIP (PointCare V2).
4
Whole Blood Collection and Cardiac Biomarker Analysis
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A total of 3 mL of whole blood was collected from the cephalic vein. Half a milliliter of blood was collected in an ethylene diamine tetra-acetic acid (EDTA) tube for automated complete blood count (CBC) analysis (Mindray BC-5000VET, Shenzhen Mindray Bio-Medical Electronics Co., Ltd., Shenzhen, China). Another 0.5 mL of blood was collected in a lithium heparinized tube for automated biochemistry analysis (Mindray BS-800, Shenzhen Mindray Bio-Medical Electronics Co., Ltd., Shenzhen, China). The remaining blood was collected in a lithium heparinized tube; then, the plasma was collected, after centrifuging at 3000 rpm for 10 min. The plasma samples were stored at −80 °C for further analysis of the cTnI level. The concentrations of cTnI were measured with the Architect STAT highly sensitive troponin I immunoassay using an Alinity™ ci-series analyzer (Abbott Laboratories, Abbott Park, IL, USA) with a detection range of 0.001–50 ng/mL.
5
Automated Blood Analysis Protocol
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Blood samples were subjected to complete blood counts obtained using a Mindray BC-5000 Vet hematology analyzer (Shenzhen Mindray Bio-Medical Electronics, Shenzhen, China).
6
Hematological Evaluation of Envenomation
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After envenomation in each study period of each group, blood samples were
collected into tubes with K3EDTA anticoagulant for determination of
the hematological parameters: red blood cells (RBC), hemoglobin (Hb), mean
corpuscular volume (MCV), leukocytes, platelets, neutrophils, lymphocytes, and
monocytes using an Auto-Hematology Analyzer (Mindray BC-5000 Vet, Mindray
Biomedical Electronics Co. Ltd. Nanshan Shenzhen, China).
collected into tubes with K3EDTA anticoagulant for determination of
the hematological parameters: red blood cells (RBC), hemoglobin (Hb), mean
corpuscular volume (MCV), leukocytes, platelets, neutrophils, lymphocytes, and
monocytes using an Auto-Hematology Analyzer (Mindray BC-5000 Vet, Mindray
Biomedical Electronics Co. Ltd. Nanshan Shenzhen, China).
7
Seasonal Variation of Blood Parameters
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The total number of blood samples were 200 (50 samples collected in each season). We measured neutrophils (Neu), basophils (Bas), haematocrit (HCT), the leukocytes (WBC), lymphocytes (Lymph), mean corpuscular hemoglobin concentration (MCHC), monocytes (Mon), eosinophils (Eos), percentage of neutrophils (Neu%), percentage of lymphocytes (Lymph%), percentage of monocytes (Mon%), percentage of eosinophils (Eos%), percentage of basophils (Bas%), erythrocytes (RBC), hemoglobin (HGB), mean corpuscular hemoglobin (MCH), coefficient of variation of red blood cell distribution width (RDW-CV), standard deviation of red blood cell distribution width (RDW-SD), platelet count (PLT), mean corpuscular volume (MCV), mean platelet volume (MPV), platelet distribution width (PDW), and platelet crit (PCT) in the blood using the five-differential blood cell analyzer BC-5000 Vet (Mindray, China).
8
Nanoparticle Toxicity Assessment in Mice
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KM mice (4-6 weeks, 25 g) were intravenously administered MLipRIR NPs (100 µL, equivalent R162 concentration at 0.5 mg/kg, in saline). On day 1, 3, 5, 7 and 14, blood was collected and analyzed using a hematology analyzer (BC-5000Vet, Mindray, China). Primary blood indices, including white blood cells (WBCs), platelets (PLTs), RBCs, hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), lymphocytes, granulocytes and monocytes, were recorded to evaluate systemic toxicity.
9
Evaluating Vascular Permeability and Leukocyte Migration in CLP-Operated Mice
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CLP‐operated mice were injected with PTMT‐T cell intravenously. After 12 h, 1% Evans blue dye solution in normal saline was injected intravenously into each mouse. Thirty minutes later, the mice were killed, and the peritoneal exudates were collected after being washed with normal saline (5 mL) and centrifuged at 200 × g for 10 min. The absorbance of the supernatant was read at 650 nm. The vascular permeability was expressed in terms of dye (μg per mouse), which leaked into the peritoneal cavity according to a standard curve of Evans blue dye. For assessment of leukocyte/neutrophil migration, CLP operated mice were treated with PTMT‐T cell after CLP surgery 12 h. The mice were then sacrificed and the bronchoalveolar lavage (BAL) were washed with 0.8 mL of normal saline.[35 ] BAL fluid (200 µL) was counted by auto hematology analyzer (Mindray, BC 5000 Vet) at the Chiral Material Core Facility Center of Sungkyunkwan University. The results were expressed as leukocyte/neutrophil × 105 per BAL fluid.
10
Bronchoalveolar Lavage Fluid Analysis
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Twenty-four hours after the last challenge, each mouse was weighed and injected with 2% pentobarbital sodium (Sigma-Aldrich, St. Louis, MO, USA) at a dose of 50 mg/kg; to collect BALF, 1.6 mL of PBS was injected into the lungs in two equal portions and repeated three times by gentle aspiration, with a fluid recovery of 80% considered satisfactory. The cell suspension was centrifuged (1500 rpm, 4 °C, 10 min), and leukocytes in BALF were counted using a whole blood analyzer (BC-5000Vet, Mindray Co.
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