Washing and Feeding of Double Coverslip Cultures, Needle, Forceps, Balanced Salt Solution

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	Washing and Feeding of Double Coverslip Cultures - Washing and feeding involves the following steps: (i) remove seal using razor blade and remove large coverslip with small one still attached and flip it over, so that culture is uppermost in orientation; (ii) detach small coverslip from the large coverslip using needle 'and forceps and transfer it to a Columbia staining dish (watch glass or Petri dish may also be used) containing balanced salt solution (BSS);
iii) remove small cover slip treated as above (one at a time) and place it on a large coverslip with culture up; while removing small coverslip from dish with BSS, the amount of BSS brought with it may be controlled by the rate of removal (too much liquid will interfere in feeding operation and too little will allow air bubbles) (iv) feed the culture by adding a drop of feeding solution (e.g. BSS : serum: 50% embryo extract = 1:1:1) to the small coverslip; (v) attach a dean depression slide, using petroleum jelly, as done earlier.