The location of the ZnPc-EVs inside cells was investigated by seeding 3000 MC38 cells in an 8-chamber polystyrene vessel tissue culture-treated glass slides (Corning) and allowing them to attach overnight. This was followed by incubation for 6 and 24 h with the ZnPc-EVs with a concentration calibrated at 4 µM ZnPc. After this time, the cells were washed 3 times in PBS and fixed in PBS containing 1% formalin (J.T. Baker) at 4 °C for 20 min. The cells were then washed three times in PBS and stained with 0.25 µM DAPI (Sigma), after which the coverslips were mounted on the glass slides using Mowiol mounting medium (Sigma-Aldrich) with 2.5% w/v DABCO (Merck) and sealed with nail polish. The slides were then imaged on a Leica SP8 fluorescence microscope.
8 chamber polystyrene vessel tissue culture treated glass slides
Manufactured by Corning
Sourced in United States
The 8-chamber polystyrene vessel tissue culture-treated glass slides are a laboratory equipment product designed for cell and tissue culture applications. The product provides a standardized platform with eight separate chambers for culturing and observing cells, tissues, or other biological samples. The glass slides are treated to promote cell attachment and growth, making them suitable for a range of cell culture experiments and applications.
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2 protocols using 8 chamber polystyrene vessel tissue culture treated glass slides
1
Visualizing ZnPc-EV Cellular Localization
2
Neutrophil Degranulation Imaging Assay
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Freshly isolated human neutrophils (0.2 × 106 cells/mL in RPMI) were distributed in duplicates into 8-chamber polystyrene vessel tissue culture-treated glass slides (Corning, Big Flats, NY, USA). PMA was used at a final concentration of 50 nM, and slides were incubated at 37°C (5% CO2) for different time periods. After incubation, supernatants were removed and samples were washed 3× with PBS. 100 nM of β-tryptase labeled with Alexa-488 (in PBS) was added, followed by incubation for 15 min (37°C; 5% CO2). Supernatants were removed and samples were washed 3× with PBS, fixed with 4% paraformaldehyde, and stained with NucBlueTM live ReadyProbesTM. Samples were washed 3× with PBS and mounted using ProLongTM diamond antifade mountant. Images were acquired using 63× NA 1.40 oil objective on Zeiss LSM700 confocal microscope. Images were analyzed using Imaris software (Bitplane, Zurich, Switzerland).
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