Cell culturing involves frequent media replacement to provide nutrition to growing cells. In a standard 96 well ultra low cell attachment plate, media aspiration or dispensing has to be done extremely carefully to avoid disturbing the unattached spheroid, making this a time consuming operation. With the introduction of PrimeSurface® 96 Slit Well Plate, media exchange for 96 well plates can be efficiently handled with one step dispensing or aspiration for all 96 wells decreasing the pipetting time by over 80% while minimizing the risk of spheroid damage.
PrimeSurface® 96 Slit-well Plate
With the introduction of PrimeSurface® 96 Slit-well Plate, media exchange for 96 well plates can be efficiently handled with one step dispensing or aspiration for all 96 wells decreasing the pipetting time by over 80% while minimizing the risk of spheroid damage.
A new design of ultra-low attachment 3D plate to facilitate easy handling of media exchange without disrupting spheroid formation.
- Generate and maintain uniform spheroids
- Exchange media without disturbing spheroid formation
- Minimize media exchange time by simultaneous delivery of cell culture media to all 96 wells.
Instructions for using PrimeSurface® 96 Slit-well Plate
- This instruction explains the basic operating procedure for PrimeSurface® 96 Slit-well Plate.
- Instructions recommended here are the best operating protocols but depending upon your cell line and assay, some optimizations may be required.
- The product and instruction are subject to change.
- This product is for research purpose only.
- PrimeSurface® 96 Slit-well Plate, is designed to form and maintain uniform spheroid for long term cultures.
- New and innovative design of plate with slits on the upper half of each well.
- Media aspirating and dispensing is efficiently performed from the corners of the plate while keeping the spheroid formation undisturbed in wells.
- The product development is supported by AMED*. *Japan Agency for Medical Research and Development
- Figures below are schematic for explaining the use of PrimeSurface® 96 Slit-well Plate in spheroid formation and media exchange.
- Procedures of cell spheroid formation (Figs. 1-3) is described in detail on page 2.
- Procedure for exchanging culture media (Fig. 4) is described on page 2.
Spheroid formation procedure using PrimeSurface® 96 Slit-well Plate
I. Procedures of cell spheroid formation
- Open the packaging of a new Slit-well Plate under the hood.
- Seed cells in the plate at a density of 6,000-9,000 cells/100 μL/well.
-The recommended volume of the above cell suspension is approximately 100 μL.
-As the maximum capacity of each well is 100 μL, do not exceed 100 μL volume for cell suspension per well.
-The formed spheroids are apt to slip through the slit or protrude outside the well if the number of seeded cells is insufficient.
-Dispense the cell suspension at the bottom of each well (section b). (Do not dispense it in section a of the well, i.e. in the slit section).
- Culture the cells in the incubator.
-After every 24-48 hrs, check for spheroid formation. Gently and carefully handle the plate during this first 24-48 hrs, as the spheroids being formed in the initial stage are fragile and can break apart.
- Dispense fresh culture media (20 mL per plate) at the corner of the plate using the 25 mL pipette.
-The total volume of the dispensed culture media is 30 mL per plate, i.e. an aliquot of 10 mL during cell seeding, plus 20mL in this step (as stated above).
-Maximum capacity of the plate is 40 mL/plate.
-Dispense media slowly (should take about 20 seconds to dispense 20 mL volume to the plate).
- Gently shake the plate back and forth in all directions couple of times to ensure that all the wells have been evenly
covered with media.
- Place the plate in the incubator.
II. Procedure of exchanging culture media
- Take the plate out of the incubator and check the spheroid formation under a microscope before exchanging the
- Aspirate the media from a corner of the plate with a 25 mL pipette and dispose of it.
-Apply the pipette tip to the corner of the plate and gently aspirate the old culture media while slightly tilting the plate.
-The plate must be tilted in order to properly aspirate the culture media.
-The recommended aspirating volume is 15 to 20 mL (takes about 20 seconds).
- Dispense fresh media from the corner of the plate with a 25 mL pipette. This volume should be the same as that
of aspiration step (2).
-Note that the capacity of the slit plate is 40 mL/plate. It is advised to stay with total volume of 30 mL/plate.
-Dispense media slowly (takes 20 sec) to avoid any overflow.
- Gently shake the plate back and forth in all directions couple of times to ensure that all the wells and corners of the
plate have been evenly covered with media.