Biotek synergy htx multi mode microplate reader

SARS-CoV-2 spike pseudovirus was produced using the third-generation lentiviral packaging plasmids as described previously [34 (link)]. The transfer plasmid encoding luciferase and ZsGreen (BEI Resources, catalog #NR-52516), helper plasmid encoding Gag/pol (BEI Resources, catalog #NR-52517), Tat (BEI Resources, catalog #NR-52518), Rev (BEI Resources catalog #NR-52519), and plasmid encoding spike of SARS-CoV-2 Wuhan (BEI Resources catalog #NR-52514) (Addgene, Watertown, MA, USA) were transfected in HEK 293T cells propagated in DMEM with 10% FBS. The pseudovirus-containing supernatants were collected after 48 h of transfection and filtered through 0.45 µm membrane filters, and aliquots were stored at −80 °C until further use. The infectivity titer of SARS-CoV-2 spike pseudovirus was determined using 293T cells overexpressing the human ACE2 receptor. Briefly, the 293T cells overexpressing human ACE2 were infected with 10-fold serial dilutions of pseudovirus in a 96-well white/clear bottom plate (Thermo Fisher Scientific, Waltham, MA, USA, catalog #165306). At 72 h post infection, the plates were equilibrated to room temperature, and 100µL of BrightGlo luciferase assay reagent (Promega, Madison, WI, USA, catalog #E2620) was added to each well. The luminescence was measured using Luminometer (BioTek Synergy HTX Multi-Mode Microplate Reader, Agilent, Santa Clara, CA, USA).

To examine
the yeast cell-killing efficacy of the generated NFAP γ-core
peptide derivatives, the growth ability of C. albicans SC5314 was monitored in the presence of 0.5× MIC, 1× MIC,
and 2× MIC of the peptides that proved to be effective in the
antifungal susceptibility tests. The experiment was conducted in a
flat-bottom 96-well microtiter plate (TC Plate 96 Well, Suspension,
F; Sarstedt, Nümbrecht, Germany): 100 μL of OD₆₀₀ = 0.2 mid-log phase cells prepared in LCM were mixed with 100 μL
of 1× MIC, 2× MIC, or 4× MIC of the peptide diluted
in LCM. The plates were incubated statically at 30 °C for 24
h. The absorbance (OD₆₀₀) of each well was then measured
after shaking for 5 s every hour using a microtiter plate reader (BioTek
Synergy HTX Multi-Mode Microplate Reader; Agilent Technologies, Santa
Clara, CA, USA). Untreated cells (100 μL LCM mixed with 100
μL OD₆₀₀ = 0.2 mid-log phase cells prepared in LCM)
served as growth controls. To examine the viability of the treated C. albicans cells, 5–5 μL cultures from
each well were dropped onto the YPD agar plate after the last OD₆₀₀ measurement and allowed to dry before incubation at 30
°C for 24 h. The plates were photographed (Versa Doc Imaging
System 4000 MP; Bio-Rad, Hercules, CA, USA). These experiments were
repeated two times, involving three technical replicates.