RESUMO
OBJECTIVE: To evaluate the effect of different types of microplate and loading volumes on the detection results of multi-function microplate reader, and to optimize the analysis method. METHODS: A multi-function microplate reader was used to perform spectrum scanning on each of 5 detection holes of common and ultraviolet(UV) microplates, and the applicable detection wavelength range was those with light transmittance greater than 80.00%. The optical density measurement was carried out on each 12 detection holes of common and UV microplates at different wavelengths, then the matching of the detection holes was compared. Potassium permanganate was quantitatively analyzed by common microplate and UV microplate, while acetone was analyzed by UV microplate, and then detection limit, lower limit of quantitation(LLQ), accuracy and precision at different loading volumes and concentrations were calculated and compared. RESULTS: The shortest applicable analyzing wavelengths for common and UV microplates were(362±2) and(230±3) nm respectively, while the longest applicable analyzing wavelengths were both 1 000 nm. The light transmittance of UV microplate was higher than that of common microplate when the analyzing wavelengths were lower than 400 nm(P<0.01). The deviation and range of light transmittance of detection holes analyzed by UV microplates were smaller than that of common microplates when the analyzing wavelengths were 350-1 000 nm(P<0.05). The detection limit and LLQ of potassium permanganate by multi-function microplate reader was not associated with the types of microplate. The adding standard recoveries of potassium permanganate by UV microplate was higher than that by common microplate(P<0.05). The adding standard recoveries of potassium permanganate by loading volumes of 200 and 250 μL was lower than that by loading volumes of 150 μL(P<0.01), while adding standard recovery of acetone by loading volumes of 200 μL was lower than that by loading volumes of 150 μL(P<0.05). CONCLUSION: When using a multi-function microplate reader to detect chemicals, it is recommended to use UV microplate with wavelengths at the range of 230-1 000 nm, and loading volumes of 200-250 μL.
RESUMO
OBJECTIVE: To compare the effects of different anesthetics and blood sampling methods on blood routine test results in experimental animals. METHODS: A total of 42 specific pathogen free( SPF) male Sprague Dawley( SD) rats and 59 SPF male Kunming( KM) mice were randomly divided into 4 groups( control group,ether group,chloral hydrate group and pentobarbital sodium group). Ether group animals were treated with ether inhalation anesthesia; animals in chloral hydrate group and pentobarbital sodium group were injected intraperitoneally with chloral hydrate or pentobarbital sodium. The control group received no anesthesia treatment. Blood samples were collected by different ways: orbital venous plexus,abdominal aorta or eyeball enucleation. White blood cell( WBC) count,red blood cell( RBC) count,platelet(PLT) count,hemoglobin(Hb) level and hematocrit(HCT) in blood samples were analyzed. RESULTS: The RBC count,Hb level and HCT of SD rats in pentobarbital sodium group were significantly lower than those in control group( P <0. 05). The HCT of SD rats in ether group was lower than that in control group( P < 0. 05). The WBC count of orbital venous plexus of KM mice was lower than that taken by eyeball enucleation in control group( P < 0. 05),but the WBC count of orbital venous plexus was higher than that taken by eyeball enucleation in chloral hydrate group( P < 0. 05). The RBC count,Hb level,HCT of KM mice in pentobarbital sodium group were significantly lower than those in control group(P < 0. 05). CONCLUSION: The anesthetic can affect the blood routine test results of experimental animals. Different blood sampling methods have effects on blood routine test results of KM mice.