With cotton rolls, isolation of the selected sites was done, and drying with air, supragingival plaque was removed gently not contacting marginal gingiva to prevent contamination or blockage of the microcapillary pipette. Samples contaminated with blood or saliva were discarded. GCF was collected by placing white color-coded 1–5 μL calibrated microcapillary pipettes (Sigma-Aldrich Chemical Company, St. Louis, USA). Using extracrevicular (unstimulated) method, 1 μL of GCF was collected, which was considered as standard volume [Figure 1 ].
Microcapillary pipette
Manufactured by Merck Group
Sourced in United States
Microcapillary pipettes are precise liquid handling instruments designed to accurately measure and dispense small volumes of liquids, typically in the range of microliters. They feature a narrow, elongated tip that can access small sample sizes and facilitate the transfer of minute quantities of fluids.
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Lab products found in correlation
5 protocols using microcapillary pipette
1
Gingival Crevicular Fluid Collection Protocol
2
Gingival Crevicular Fluid Collection
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Tooth polishing was performed before the start of the study. With a sterile mouth mirror and a Goldman/Fox Williams periodontal probe, the periodontal status was examined clinically. After isolating the area with cotton rolls, 1–3 μl microcapillary pipettes (Sigma Aldrich Chemical Company, USA; Catalog No. p0549) were used for GCF collection. By positioning the tip of the pipette extracrevicularly (unstimulated) for 30 s, a standard volume of 3 μl GCF was collected from each test site, i.e., buccal, mesial, distal, and lingual/palatal sites of matured primary molars in primary dentition, using the graduation marks on the micropipette, as described by Ureles et al. The GCF in mixed dentition was collected from sites distal to second primary molar, and in permanent dentition distal sites of permanent first molars as mentioned by Rody et al [Figure 1 ].[9 (link)] The pipettes were transferred to the laboratory immediately and frozen at −70°C for analysis.
3
Gingival Crevicular Fluid Collection
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GCF samples were collected a day after the probing depth was recorded. GCF was collected from the deepest probing site after removal of supra-gingival plaque. The site of collection was isolated to prevent contamination of saliva. A 5-μL sample of GCF was collected by placing microcapillary pipettes (Sigma Chemical Company, St Louis, MO, USA) at the entrance of gingival crevice. The pipettes were placed at the entrance of the sulcus and there was no stimulation. The samples of GCF were stored in plastic vials at -70 0 C until analyzed for LXA4 concentrations.
4
GCF Sampling and Storage Protocol
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GCF samples of 5 μl were collected from the sites receiving treatment using a graduated Microcapillary pipette by extracrevicular method. The samples from the treatment sites were collected at baseline, 7 days after laser irradiation, and 21 days after retraction. These pipettes were wrapped in tin foil, transferred to plastic vials, and stored at −70°C until the time of assay. Microcapillary pipette was procured from Sigma Aldrich, Bangalore, India. The storage of GCF samples was done at Coorg Institute of Dental Sciences.
5
Organotypic Slice Culture and Cell Transplantation
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All procedures were approved by the Institutional Animal Care and Use Committee of Seoul National University. The organotypic slice culture was performed as previously described27 (link)–29 (link). Cells (6 × 103 cells/μl) prepared in neurobasal medium were transplanted onto the slice using aspirator tube assembly for microcapillary pipette (Sigma). In 7 days, the slices were fixed with 4% PFA overnight at 4°C, permeabilized and blocked with 0.1% Triton X-100 in 3% BSA. They were incubated overnight at 4°C with mouse anti-SOX2, 1:500, rabbit anti-TuJ1, 1:1000 and mouse anti-human nuclei, 1:500 (Millipore). Incubation with the secondary antibodies was performed as described above. The z-stack images were captured using a laser-scanning confocal microscope at 3–4 µm intervals.
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