Nunc 96 well polypropylene microwell plate

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Nunc™ 96-well polypropylene MicroWell™ plate is a laboratory equipment product designed for a variety of applications. It features a 96-well format constructed from polypropylene material.

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Lab products found in correlation

Low tox h rabbit complement, by Cedarlane (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions) Sorvall legend rt, by Thermo Fisher Scientific (1 mentions)

3 protocols using nunc 96 well polypropylene microwell plate

Biofilm Maturation and Antimicrobial Evaluation

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To develop a mature biofilm, 10⁶ CFU/ml of each strain was resuspended in saline (B. Braun, Germany). Next, 100 μL of this suspension were incubated at 37°C and 5% CO₂ for 90 min in static conditions in a Nunc™ 96-well polypropylene MicroWell™ plate (Thermo Fisher Scientific, USA). After incubation, each well was rinsed twice with 100-μL saline and 150-μL tryptic soy broth with 0.5% glucose with or without (positive control) each extract at different concentrations (32, 16 and 8 mg/mL, respectively) and, thereafter, incubated at 37°C and 5% CO₂ for 24 h. Following incubation, each well was rinsed twice with 100-μL saline and 150-μL tryptic soy broth with 10% alamarBlue (BIO-RAD) (Pettit et al., 2005 (link)) and, then, incubated at 37°C and 90 rpm for 30 min (Peeters et al., 2008 (link)). After incubation, the fluorescence was measured using an excitation wavelength of 560 nm and an emission wavelength of 590 nm. This experiment was performed in eight wells per extract in triplicate for each strain (n = 24).

Aguilera-Correa J.J., Nohynek L., Alakomi H.L., Esteban J., Oksman-Caldentey K.M., Puupponen-Pimiä R., Kinnari T.J, & Perez-Tanoira R. (2023). Reduction of methicillin-resistant Staphylococcus aureus biofilm growth and development using arctic berry extracts. Frontiers in Cellular and Infection Microbiology, 13, 1176755.

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Complement-Dependent Cytotoxicity Screening

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14 of the 64 flow cytometry samples were screened by complement-dependent cytotoxicity. 7 had class II HLA sensitization and 7 lacked allosensitization. The CDC assay was performed as described by Diaz et al., with minor modifications(9 (link)). Briefly, Cells were added to each well with 1×10⁵ in 25 ul HBSS of Nunc^™ 96-Well Polypropylene MicroWell^™ Plate (Thermo Fisher Scientific, Waltham, MA), and incubated at 4°C for 30min with 25 ul/well of human sera treated with or without DTT (2.5mM in final) for 30min at 37°C. The cells were washed at 3 times with HBSS, then treated with 50ul/well of 11-fold dilution of Low-Tox®-H Rabbit Complement (Cedarlane, Burlington, NC) for 90min at 37°C. The cells were stained with FDA (0.5 ug/ml)/PI (2.5mg/ml, Sigma-Aldrich) at 4°C for 15min. Data were collected using a BD Accuri C6 Flow Cytometer and software (BD Accuri, Ann Arbor, MI, USA).

Ladowski J.M., Martens G.R., Reyes L.M., Wang Z.Y., Eckhoff D.E., Hauptfeld-Dolejsek V., Tector M, & Tector A.J. (2018). Examining the Biosynthesis and Xenoantigenicity of Class II SLA Proteins. Journal of immunology (Baltimore, Md. : 1950), 200(8), 2957-2964.

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Evaluating RBC Lysis by Polyplexes

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RBC lysis in the presence of polyplexes was evaluated to determine their hemocompatibility. Blood was collected from healthy human volunteers in tubes containing potassium EDTA (Sigma-Aldrich). Collected whole blood was centrifuged at 1,500 × g for 10 min at RT (Sorvall Legend RT; Thermo Scientific, Waltham, MA), and RBCs were washed three times with PBS. To determine hemolysis, RBCs were diluted six times with PBS and incubated at RT with polyplexes (polymer:pDNA ratio 2:1 w/w) for 3 hr. The RBC-polyplex mixture was centrifuged for 10 min, and the supernatant (50 μL) was dissolved in 150 μL of 40:1 (v/v) ethanol: HCl mixture in a Nunc 96-well polypropylene MicroWell plate (Thermo Scientific, Waltham, MA). The absorbance was measured at 399 nm. RBCs incubated with deionized water were used as the positive control for complete lysis, and RBCs incubated with PBS served as no lysis control.

Joshi C.R., Raghavan V., Vijayaraghavalu S., Gao Y., Saraswathy M., Labhasetwar V, & Ghorpade A. (2018). Reaching for the Stars in the Brain: Polymer-Mediated Gene Delivery to Human Astrocytes. Molecular Therapy. Nucleic Acids, 12, 645-657.

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