Growth of cells was assayed using the Promega CellTiter 96 Cell Proliferation Assay. Cells were resuspended to a final concentration of 1.0 x 105/mL in RPMI. 50 μl of the cell suspension (5,000 cells) was added to each well of the 96-well plate containing 50 μl of media with corresponding treatment resulting in a total volume of 100 μl. The micro plates were incubated at 37°C for 24–72 hours in a humidified, 5% CO2 atmosphere. Per manufacturer’s instructions, following incubation, 20 μl of MTS / PMS solution was added to each well and incubated for 4 hours. Absorbances were read at 490 nm using the Synergy H1m multimode micro plate reader.
Synergy h1m
Manufactured by Synergy Software
The Synergy H1M is a multi-mode microplate reader capable of absorbance, fluorescence, and luminescence detection. It features a high-performance monochromator system and a sensitive photomultiplier tube detector. The Synergy H1M is designed for general quantitative and qualitative analysis in life science research and development applications.
Automatically generated - may contain errors
7 protocols using synergy h1m
1
Cell Proliferation Assay Protocol
2
Electroporation and Fluorescence Quantification
3
Cell Proliferation Assay with CCK-8
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Here, 2 × 103 cells per well were seeded into 96‐well plates and cultured for 4 d. Live cell numbers were determined using a cell counting kit CCK‐8 (TransGen Biotech). Absorbance at 450 nm was measured using a microplate reader (Synergy H1M).
4
Cytotoxicity Assay of GL261 Cells
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Here, 4 × 103 GL261 cells (T) per well were seeded into 96‐well plates and cocultured with murine splenocytes (E) at the E/T ratio of 200:1. After 4 h, lysate of GL261 cells was detected using an LDH Cytotoxicity Assay Kit. Absorbance at 490 nm was measured using a microplate reader (Synergy H1M).
5
Photothermal Cytotoxicity Assay of Nanoparticles
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MDA-MB-231
cells were seeded into 96-well plates at 50 000 cells/well,
treated with media containing 0 or 8 × 109 NS–PS/mL
for 24 h, and then subjected to no irradiation, or irradiation for
5 min/well using the 800 nm pulsed laser with a beam diameter of approximately
1 cm (to cover the entire well) and a power density of 1 W/cm2 or using the LED lightbox with a 500 nm longpass filter.
The plate was prewarmed and kept at 37 °C in a digital 2 block
heater (VWR) until the end of irradiation. After irradiation, the
cells were incubated for 24 h, then the media was removed and an Alamar
blue viability reagent (Thermo Fisher, Waltham, MA, USA; diluted 1:10
in complete cell culture media) was added per manufacturer recommendations.
Sample fluorescence was measured on a Synergy H1M plate reader with
excitation and emission wavelengths of 560 and 590 nm, respectively.
To analyze the data, background (Alamar blue reagent without cells)
was subtracted from the fluorescence reading in each well.
cells were seeded into 96-well plates at 50 000 cells/well,
treated with media containing 0 or 8 × 109 NS–PS/mL
for 24 h, and then subjected to no irradiation, or irradiation for
5 min/well using the 800 nm pulsed laser with a beam diameter of approximately
1 cm (to cover the entire well) and a power density of 1 W/cm2 or using the LED lightbox with a 500 nm longpass filter.
The plate was prewarmed and kept at 37 °C in a digital 2 block
heater (VWR) until the end of irradiation. After irradiation, the
cells were incubated for 24 h, then the media was removed and an Alamar
blue viability reagent (Thermo Fisher, Waltham, MA, USA; diluted 1:10
in complete cell culture media) was added per manufacturer recommendations.
Sample fluorescence was measured on a Synergy H1M plate reader with
excitation and emission wavelengths of 560 and 590 nm, respectively.
To analyze the data, background (Alamar blue reagent without cells)
was subtracted from the fluorescence reading in each well.
6
MTS Assay for Cell Proliferation
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Growth of cells were assayed using the Promega CellTiter 96 Cell Proliferation Assay. Cells were resuspended to a final concentration of 1.0×105/mL in RPMI. 50 µl of the cell suspension (5,000 cells) was added to each well of the 96-well plate containing 50 µl of media with corresponding treatment resulting in a total volume of 100 µl. The microplates were incubated at 37°C for 24–72 hours in a humidified, 5% CO2 atmosphere. Per manufacturers instructions, following incubation, 20 µl of MTS solution was added to each well and incubated for 4 hours. 100 µl of stop solution was added to each well and incubated for 1 hour. Absorbances were read at 570 nm using the Synergy H1m multimode microplate reader.
7
Fluorescence Spectroscopy of Nanodiscs
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NBD- and OG-labeled proteins (10 nM) were incubated with nanodiscs at the indicated concentrations in reconstitution buffer. The fluorescence spectrum of samples was collected on a Synergy H1M plate reader with excitation at 460 nm and emission from 500–650 nm.
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