[A glass micropipette vacuum technique of cerebrospinal fluid sampling in C57BL/6 mice].

The purpose of this study was to establish a suitable method for extracting cerebrospinal fluid (CSF) from C57BL/6 mice. A patch clamp electrode puller was used to draw a glass micropipette, and a brain stereotaxic device was used to fix the mouse's head at an angle of 135° from the body. Under a stereoscopic microscope, the skin and muscle tissue on the back of the mouse's head were separated, and the dura mater at the cerebellomedullary cistern was exposed. The glass micropipette (with an angle of 20° to 30° from the dura mater) was used to puncture at a point 1 mm inboard of Y-shaped dorsal vertebral artery for CSF sampling. After the first extraction, the glass micropipette was connected with a 1 mL sterile syringe to form a negative pressure device for the second extraction. The results showed that the successful rate of CSF extraction was 83.33% (30/36). Average CSF extraction amount was (7.16 ± 0.43) µL per mouse. In addition, C57BL/6 mice were given intranasally ferric ammonium citrate (FAC) to establish a model of brain iron accumulation, and the CSF extraction technique established in the present study was used for sampling. The results showed that iron content in the CSF from the normal saline control group was not detected, while the iron content in the CSF from FAC-treated group was (76.24 ± 38.53) µmol/L, and the difference was significant. These results suggest that glass micropipette vacuum technique of CSF sampling established in the present study has the advantages of simplicity, high success rate, large extraction volume, and low bleeding rate, and is suitable for the research on C57BL/6 mouse neurological disease models.

A "turn-on" sensor based on MnO2 coated UCNPs for detection of alkaline phosphatase and ascorbic acid.

Alkaline phosphatase (ALP), an extraordinary enzyme, can catalyze the dephosphorylation of small molecules, proteins and nucleic acids. ALP has been confirmed as a crucial serum diagnostic indicator, since the abnormal level of ALP is closely related to a variety of pathological processes, especially in the early diagnosis and screening of cancer. Herein, we designed a "turn-on" sensor to detect ALP and ascorbic acid (AA) based on the redox reaction between manganese dioxide (MnO2) coated upconversion nanoparticles (UCNPs) and AA. Firstly, 2 nm sized NaYF4:Yb3+,Tm3+ UCNPs were synthesized by a facile one-pot hydrothermal method. Then MnO2 coated UCNPs were prepared via electrostatic interactions between MnO2 nanosheets and UCNPs. MnO2 nanosheets could absorb blue light emitted by UCNPs near 471 nm under laser excitation at 980 nm, and so the luminescence of UCNPs was quenched based on luminescence energy transfer (LET). In the presence of AA, the luminescence was recovered again by the redox reaction between AA and MnO2 coated UCNPs. MnO2 nanosheets were reduced to Mn2+ and UCNPs were released. Furthermore, the "turn-on" sensor was applied to monitor ALP since the phosphate group of 2-phospho-l-ascorbic acid (AAP) was removed by ALP to yield AA. The bio-assay showed a good linear relationship between the restored luminescence intensities (ΔI = I-I0) and ALP concentrations ranging from 0.25 to 150 mU mL-1 with a limit of detection (LOD) of 0.045 mU mL-1 and between ΔI and AA concentrations ranging from 0.5 to 250 µM with an LOD of 0.29 µM. The luminescent sensor was also successfully applied for detection of ALP and AA in human serum samples with recoveries from 94.9% to 104.6% and 99.4% to 104.9%.