96 well glass bottom plates

Manufactured by Corning

Sourced in United States

The 96-well glass-bottom plates provide a standardized microplate format with a glass surface at the base of each well, enabling high-quality imaging and analysis of cells or other samples.

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Lab products found in correlation

Mitotracker green fm, by Thermo Fisher Scientific (3 mentions) Lysotracker green dnd 26, by Thermo Fisher Scientific (3 mentions) Er tracker, by Thermo Fisher Scientific (2 mentions) Lsm 710 axio obzerver z1 duo nlo laser scanning microscope, by Zeiss (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Ld c apochromat, by Zeiss (1 mentions) Cellmask green, by Thermo Fisher Scientific (1 mentions) Hoechst, by Thermo Fisher Scientific (1 mentions) Cytation 5 cell imager, by Agilent Technologies (1 mentions) Lionheart fx imager, by Agilent Technologies (1 mentions) Gene5, by Agilent Technologies (1 mentions)

5 protocols using 96 well glass bottom plates

Intracellular Localization of Photosensitizers

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Intracellular distribution of PS and PD was studied by using the LSM 710 Axio Obzerver Z1 DUO NLO laser scanning microscope (Carl Zeiss, Germany). The images were obtained using a LD C-Apochromat water immersion objective lens 40×/1.1. The GL261 cells were seeded in 96-well glass-bottom plates (Corning, USA) at 10⁴ cells per well and grown overnight. The cells were then incubated with 10 μM photosensitizers in serum-free culture medium for 1–4 h, followed by washing with PBS and confocal image acquisition. The fluorescence of PS and PD was excited at 633 nm and recorded in the range of 650–735 nm.
For colocalization analysis of PS and PD after 3.5 h of incubation of GL261 cells with the respective photosensitizer, the following dyes were added for 30 min (ThermoFisherScientific): 0.5 μM LysoTracker Green DND-26 for lysosomes, 0.5 μM ER-Tracker for endoplasmic reiculum, 0.5 μM MitoTracker Green FM for mitochondria, 5 μM BODIPY FL C5-ceramide complexed to BSA for Golgi apparatus. Dyes were added to living cells that had been incubated with the photosensitizers. Staining was performed according to the manufacturer’s instructions. Fluorescence of stained organelles was excited by an argon laser at 488 nm and registered in the range of 500–560 nm.

Turubanova V.D., Balalaeva I.V., Mishchenko T.A., Catanzaro E., Alzeibak R., Peskova N.N., Efimova I., Bachert C., Mitroshina E.V., Krysko O., Vedunova M.V, & Krysko D.V. (2019). Immunogenic cell death induced by a new photodynamic therapy based on photosens and photodithazine. Journal for Immunotherapy of Cancer, 7, 350.

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Intracellular Localization of Pz I and Pz III in Glioma Cells

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Glioma GL261 cells were seeded in 96-well glass-bottom plates (Corning Inc., Corning, NY, USA) at 1 × 10⁴ cells per well and grown overnight (24 h). Next, the glioma cells were incubated in a serum-free medium supplemented with 10 μM pz I or pz III for 4 h, followed by washing with phosphate-buffered saline (PBS, Gibco, Thermo Fisher Scientific, Waltham, MA, USA). To analyze the intracellular distribution of pz I and pz III, the following dyes were added 30 min before the end of the incubation period: LysoTracker Green DND-26 for lysosomes (0.5 μM), ERTracker for endoplasmic reticulum (ER, 0.5 μM), MitoTracker Green FM for mitochondria (0.5 μM), BODIPY FL C5-ceramide complexed with bovine serum albumin for Golgi apparatus (5 μM), and DAPI for nucleus (2.8 μM) (all fluorescent dyes were purchased from Thermo Fisher Scientific, Waltham, MA, USA). The stained glioma GL261 cultures were observed in an LSM 710 Axio Obzerver Z1 DUO NLO laser scanning microscope (Carl Zeiss, Oberkochen, Germany) with an LD C-Apochromat water immersion objective lens, 40×/1.1. Fluorescence of the stained organelles was excited using an argon laser at 488 nm and recorded at 500–560 nm [26 (link)].

Redkin T.S., Sleptsova E.E., Turubanova V.D., Saviuk M.O., Lermontova S.A., Klapshina L.G., Peskova N.N., Balalaeva I.V., Krysko O., Mishchenko T.A., Vedunova M.V, & Krysko D.V. (2023). Dendritic Cells Pulsed with Tumor Lysates Induced by Tetracyanotetra(aryl)porphyrazines-Based Photodynamic Therapy Effectively Trigger Anti-Tumor Immunity in an Orthotopic Mouse Glioma Model. Pharmaceutics, 15(10), 2430.

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Visualizing C. jejuni EV-induced Changes

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HeLa cells were seeded in 96-well glass bottom plates (1 × 10⁴ cells/mL; Corning) and stimulated with C. jejuni EVs (5 µg/mL) for 48 hours. After washing in PBS, cells were stained with 100 µL of 1 x CellMask Green (Thermo Fisher Scientific) and Hoechst (1:2,000; Thermo Fisher Scientific). Cells were fixed with 4% paraformaldehyde (PFA) for 5 min at 37°C and washed three times with PBS. Imaging was performed with an FV1200 Confocal Microscope (Evident Australia) using a 20×, 0.75 NA objective with excitation at 405 nm for Hoechst and 488 nm for CellMask Green. Acquisition parameters were maintained at consistent settings for all images acquired. Cell cytoplasm and nucleus areas were quantified using the cell function on Imaris software (Bitplane, Oxford instruments). The algorithm used was for nucleus and cell detection. Nuclei were detected using a smooth filter width of 1.40 µm. Cell body detection used a cell smooth filter width of 1.50 µm, expanded on the nucleus, and assumed one nucleus per cell.

Le L.H., Elgamoudi B., Colon N., Cramond A., Poly F., Ying L., Korolik V, & Ferrero R.L. (2024). Campylobacter jejuni extracellular vesicles harboring cytolethal distending toxin bind host cell glycans and induce cell cycle arrest in host cells. Microbiology Spectrum, 12(3), e03232-23.

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Photosensitizer Localization and Immunogenic Cell Death

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The localization of photosensitizers in cytoplasmic organelles during PDT plays a major role in their ability to induce ICD. To examine this, murine fibrosarcoma MCA205 cells were treated with pz I or pz III and analyzed by using a LSM 710 Axio Observer Z1 DUO NLO laser scanning microscope (Carl Zeiss, Germany). Images were acquired with a LD C-Apochromat water immersion objective lens 40x/1.1. The fibrosarcoma MCA205 cells were seeded at a density of 10⁴ cells per well in 96-well glass-bottom plates (Corning) and were grown overnight. Next, the cells were incubated with 10 μM photosensitizers in a serum-free culture medium for 4 h, followed by washing with PBS. Fluorescence was excited at 594 nm and recorded at 600–670 nm. Confocal images were acquired.
To determine the subcellular localization of these photosensitizers after they were incubated for 3.5 h with fibrosarcoma MCA205 cells, the following dyes were added for 30 min (ThermoFisherScientific) according to the manufacturer's instructions: 0.5 μM LysoTracker Green DND-26 for lysosomes, 0.5 μM ERTracker for ER, 0.5 μM MitoTracker Green FM for mitochondria, 5 μM BODIPY FL C5-ceramide complexed with BSA for Golgi apparatus, and 3,0 μM DAPI for nucleus. Excitation was done by an argon laser at 488 nm and fluorescence was registered in the range of 500–560 nm.

Turubanova V.D., Mishchenko T.A., Balalaeva I.V., Efimova I., Peskova N.N., Klapshina L.G., Lermontova S.A., Bachert C., Krysko O., Vedunova M.V, & Krysko D.V. (2021). Novel porphyrazine-based photodynamic anti-cancer therapy induces immunogenic cell death. Scientific Reports, 11, 7205.

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Time-lapse Imaging of Trypanosoma cruzi Infection

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Human foreskin fibroblasts (HFF) or Vero cells were seeded in 96 well glass bottom plates (Corning Life Sciences, NY) or 8 well 1 µ-slides (Ibidi, Fitchburg, WI) and infected with Tdtomato-expressing colombiana or CL strain parasites labeled with CellTrace Violet in ratios of 2:1 to 10:1 (parasites:host cells). For time-lapse video of amastigote replication, extracellular trypomastigotes were removed by washing and plates were placed in humid chamber with CO₂ and imaged at 40X magnification every 15 min for 48 hr in a Lionheart FX imager (BioTek, Winooski, VT). Images and time lapse videos were analyzed with the Gene5 software (BioTek). Live cell imaging of cultures at different times post-infection and after drug addition or removal were performed in a Cytation5 cell imager (BioTek) or a Delta Vision II Microscope System (GE Healthcare Biosciences, PA).

Sánchez-Valdéz F.J., Padilla A., Wang W., Orr D, & Tarleton R.L. (2018). Spontaneous dormancy protects Trypanosoma cruzi during extended drug exposure. eLife, 7, e34039.

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