Submitted by Lauren French (download pdf)

  • In the tissue culture hood, remove cover slips one-by-one with flame-sterilized tweezers.
  • Remove 3 (60 x 15 mm) Petri dishes and place in hood
  • Remove the lids of 2 of the Petri dishes.  Leave one of the open Petri dishes empty and fill      the other with 70% ethanol
  • Cut each plastic cover slip lengthwise into 4 pieces and allow the pieces to fall into the empty, open  Petri dish
  • Remove each put piece with tweezers and cut each strip again into 4 pieces allowing them to fall  into the Petri dish containing ethanol.  Repeat with all needed coverslips.  (This should make 16 cut  coverslip pieces for each original coverslip).
  • When finished cutting cover slips, allow them to soak in ethanol for 30-60 minutes with the lid on the  Petri dish
  • After soaking, open and fill the third Petri dish with 70% ethanol and obtain a few larger Petri dishes (150 x 15mm) and place them in the hood.  Remove the lids of the 150 x 15mm Petri dishes keeping them sterile.
  • Remove the coverslips one-by-one with tweezers and dip each several times in the second dish filled with 70% ethanol. Shake off excess ethanol into empty 60 x 15mm Petri dish.
  • Place each cover slip tilted against the edge of the lid of the larger Petri dish (150 x 15mm) to dry in hood. (Allow them to dry completely before proceeding to the next step.)
  • While drying, remove several 35 x 10mm tissue culture dishes and place in hood.  Also, remove collagen from refrigeration at 4°C and allow to come to room temperature.
  • When dry, transfer 6coverslips to one 35 x 10mm tissue culture dishes.
  • Pipette collagen onto one cover slip (enough to cover, about 2mL for every 6, 35 x10mm dishes) and let sit ~1 minute.  Push the coverslips down to the bottom of the dish with the tip of the pipette to ensure the coverslip has been coated using sterile technique.
  • Remove excess collagen with pipette (leaving wet) and repeat for all other coverslips.
  • Dry for a few hours in the hood.
  • When dry, place lids on each 35mm dish and transfer up to 6 dishes to a large Petri dish (150 x 15mm) and cover.
  • Wrap the Petri dish with parafilm and store at 4°C for weeks until use.  Bring to room temperature just prior to replating cells.

 

 

 

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