Instructions for use

The Picovitro plates can be loaded with cells either manually or using an automatic cell sorter.

Manual cell seeding into Picovitro plates

The cells can be manually seeded, using a standard pipette, into the Picovitro plate by random settling of the cells into each well. After cell seeding gently seal the Picovitro plate with the semipermeable membrane. For long-term single cell/clone analysis, place the Picovitro plate in a standard cell culture incubator (37ºC, 5% CO2).

Single cell sorting and seeding into Picovitro plates

For automatic single cell seeding into predefined wells, a cell sorter with an x/y stage for plate sorting can be employed. Before following steps are performed, make sure your flow cytometer is carefully aligned and adjusted for sorting applications, including correct drop delay settings etc. Contact your instrument manufacturer for information regarding plate sorting.

  1. Place the sterile Picovitro plate on an adsorbing material, e.g., facial tissue.
  2. Manually dispense culture medium onto the Picovitro plate (800 µl/1000 micro wells). Distribute the liquid with a sterile cell scraper and remove excess liquid by scraping it over (the edge of) the Picovitro plate. Each micro well should be compartmentalized from the neighboring well, containing a fixed volume of culture medium.
  3. Place the Picovitro plate in a standard microtiter plate holder in a flow cytometer with sorting capabilities.
  4. Start the automatic single cell seeding (approx 1 well/sec).
  5. When cell seeding is performed, gently pick out the Picovitro plate and seal with the semipermeable membrane. To avoid air bubble formation, 1-2 drops of culture medium can be added on top of the Picovitro plate to facilitate sealing of the Picovitro plate with the semi-permeable membrane.
  6. For long-term single cell/clone analysis, place the Picovitro plate in a standard cell culture incubator (37ºC, 5% CO2).

Detection

The Picovitro plate is fully compatible with standard detection methods, such as microscopy stages etc.

Fig.1

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Fig.2

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