Ocular hypertension was induced in Group 2 by ic injection of microbeads as previously described.34 (link) Anesthesia was induced with 5% Isoflurane (Baxter Healthcare Corp, Deerfield, IL, USA)/1.5 L per minute O2 and maintained at 3.5% throughout the procedure. Using a 15° blade (Fine Science Tools, Reading, PA, USA) a small 2-mm incision was made at the peripheral cornea and aqueous humour was allowed to exude. Using the same incision site, a 10-μL solution of microbeads was administered with a glass micropipette, produced in-house from a glass capillary rod (Harvard Apparatus, Kent, UK) using a Flaming-Brown micropipette puller (Sutter Instruments, Novato, CA, USA). The microbead solution was loaded into the microneedle immediately before injection and consisted of 5 μL of 6-μm beads (polybead polystyrene, Cat#07312; Polysciences, Inc., Warrington, PA, USA) followed by 5 μL of 10-μm beads (polybead polystyrene, Cat#17136; Polysciences, Inc.), both at concentrations of 2 × 108/mL. Administration was made slowly and the needle was retracted with a 2-minute delay to minimize leakage. Due to the variable translucency of the eye after microbead injection, reliable ERG and optical coherence tomography (OCT) measurements were not possible.
Polybead polystyrene
Manufactured by Polysciences
Sourced in United States
Polybead polystyrene is a type of laboratory equipment used for various scientific applications. It is a spherical polymer particle made from polystyrene. The core function of Polybead polystyrene is to serve as a standard for size and concentration measurements in analytical techniques.
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2 protocols using polybead polystyrene
1
Ocular Hypertension Induction by Microbeads
2
Deionized Water Microsphere Suspension Preparation
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The deionized (DI) water used in all experiments was obtained from a Barnstead D3750 Nanopure Diamond purification system (type-1 high-performance liquid chromatography grade, resistivity 18.2 MΩ·cm at 25°C). The DI water was passed through a 0.2-μm hollow-fiber filter, ensuring bacteria- and particle-free water. The pH of the resulting water was between 6.0 and 6.5.
A microsphere suspension in DI water was used for visualizing the EZ and determining its size. Two types of polymeric, non-magnetic microspheres were purchased from Polysciences, Inc.: Polybead polystyrene (PS), and Polybead carboxylate polystyrene (CPS), the latter containing surface carboxyl groups. Mean diameter of the microspheres was 2.0 μm, and bead density was ~1.05 g/cm3. The microspheres are packaged as 2.5% solids (weight/volume) aqueous suspensions containing 5.68 x 109 particles/mL. All experiments were carried out using mixtures of the microsphere suspension with DI water at a ratio of 45 μL (1 drop)/25 mL. Suspension homogeneity was achieved by means of hand or mechanical agitation. Experiments were carried out at room temperature (296°K < T < 299°K).
A microsphere suspension in DI water was used for visualizing the EZ and determining its size. Two types of polymeric, non-magnetic microspheres were purchased from Polysciences, Inc.: Polybead polystyrene (PS), and Polybead carboxylate polystyrene (CPS), the latter containing surface carboxyl groups. Mean diameter of the microspheres was 2.0 μm, and bead density was ~1.05 g/cm3. The microspheres are packaged as 2.5% solids (weight/volume) aqueous suspensions containing 5.68 x 109 particles/mL. All experiments were carried out using mixtures of the microsphere suspension with DI water at a ratio of 45 μL (1 drop)/25 mL. Suspension homogeneity was achieved by means of hand or mechanical agitation. Experiments were carried out at room temperature (296°K < T < 299°K).
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