Studies involving infectious SARS-CoV-2 were performed as previously described (73 (link)). Two-fold serial dilutions of heat-inactivated serum at an initial dilution of 1:20, were prepared in serum free media (Minimum Essential Medium containing 25 mM HEPES and 0.05 g/L Gentamicin sulfate) and incubated with an equal volume of a solution containing 200 PFU of SARS-CoV-2-mNG (mNeonGreen) (74 (link)) for 1 h at 37°C. Virus-serum mixtures were then added to Vero E6 monolayers in 96-well optical black plates (Thermo Scientific) and incubated at 37°C. Plates were read using the BioTek Cytation 7 plate reader (EX 485 nm, EM 528 nm) at 48 h postinfection. Following background signal correction, virus neutralization half-maximal inhibitory dilution (ID50) values were determined using a 4-parameter logistic regression.
96 well optical black plates
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The 96-well optical black plates are a type of laboratory equipment designed for optical measurements. These plates feature 96 individual wells, each with a black color, which helps to minimize background interference and improve signal-to-noise ratio in various experimental applications.
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Lab products found in correlation
Celltiter glo luminescent cell viability assay kit, by Promega (4 mentions)
Synergy h4 hybrid multi mode microplate reader, by Agilent Technologies (4 mentions)
Ir700dx nhs ester, by LI COR (4 mentions)
Ivis lumina xr in vivo imaging system, by PerkinElmer (3 mentions)
Axio observer fluorescent microscopy system, by Zeiss (2 mentions)
Vero e6, by Thermo Fisher Scientific (1 mentions)
Cytation 7 plate reader, by Agilent Technologies (1 mentions)
Liquid handling system, by Tecan (1 mentions)
Jwh 133, by Cayman Chemical (1 mentions)
Mda mb 231 human breast cancer cells, by American Type Culture Collection (1 mentions)
Cary 100 bio uv vis spectrophotometer, by Agilent Technologies (1 mentions)
Apoptag in situ apoptosis detection kit, by Merck Group (1 mentions)
Cary eclipse fluorescence spectrophotometer, by Agilent Technologies (1 mentions)
Lct premier mass spectrometer, by Waters Corporation (1 mentions)
Avance 3 400 mhz system, by Bruker (1 mentions)
Avance 3 400 mhz, by Brucker (1 mentions)
6 protocols using 96 well optical black plates
1
SARS-CoV-2 Virus Neutralization Assay
2
Proton Flow Measurement in E. coli
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LR1 cells were grown in the same conditions as DH10B cells. After 3 h of secondary culture, protein expression was induced by adding 100 µM IPTG at 37 C for 2 h. The cells were then washed twice and dissolved in buffer (200 mM Na HPO , 0.9% NaCl adjusted to pH 7.6 with 0.1 M citric acid, 0.9% NaCl) to an optical density of 0.25 at 600 nm. 200 µL of dissolved cells were put in 96-well optical black plates (Thermo Scientific, Waltham, MA, USA) with 30 µL buffer (or buffer + Drug/DMSO). 230 µL buffer was used for baseline reduction. Uninduced bacterial culture was used as a control. Fluorescent measurements were carried out after adding 70 µL of citric acid (300 mM, 0.9% NaCl) by a liquid handling system (Tecan; Männedorf, Switzerland). The plate reader was set for 390 nm and 466 nm excitation filters paired with a 520 nm emission filter. The wells’ emissions were read out, alternating between two filter pairs for 60 s. The proton flow was calculated according to the procedure described previously [38 (link),49 (link)]. All experiments were performed in duplicates or triplicates.
3
Photodynamic Therapy for Cancer
4
Breast Cancer Cell Analysis Protocol
5
Photosensitizer-based Cell Viability Assays
6
Synthesis and Characterization of Fluorescent Probe
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All solvents used are of ACS or HPLC grade (Fisher Scientific, Pittsburgh, Pennsylvania). The PS, IR700DX-NHS ester, was purchased from LI-COR Bioscience (Lincoln, Nebraska). 1 H and 13 C NMR spectra were recorded at 25°C on the Brucker Avance III 400 MHz. Unless otherwise specified, chemical shifts δ were expressed in parts per million (ppm) based on tetramethylsilane (TMS) in CDCl 3 (δ 0.00 ppm), residue dimethyl sulfoxide (DMSO) (δ 2.50 ppm), or residue CHCl 3 (δ 77.16 ppm), and coupling constants J are given in Hz. Coupling patterns are abbreviated as s (singlet), d (doublet), t (triplet), q (quartet), quin (quintet), dd (doublet of doublets), ddd (doublet of doublet of doublets) and m (multiplet). Electrospray mass spectra were recorded on SYNAPT G2S TOF-MS mass spectrometer (Waters, Milford, Massachusetts) in positive ion mode. 7-Diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM) was obtained from Fischer Scientific. The following instruments, supplies and assay kits were used for in vitro studies: Synergy™ H4 Hybrid Multi-Mode Microplate Reader (BioTek, Winooski, VT), 96-well optical black plates (Fisher Scientific), CellTiter-Glo Luminescent Cell Viability Assay kit (Promega, Madison, Wisconsin).
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