BIOL 2020 Lecture Notes - Electronic Body Music, Trypsin, Dimethyl Sulfoxide

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28 Aug 2012
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[ protocol for cell labeling and injection ]- Cell labeling (cmtmr/ cmfda) and preparation of cells for injection. Day 0: place egm-2mv, ebm, and pbs in water bath, prepare staining solution: For each flask: 90 ul of cmfda solution is needed for each 5ml of ebm. Incubate in incubator for 45min: wash with pbs (aspirate the staining solution). Add 10ml egm-2mv: place in incubator at least 24hr. Wash stain wash medium: place egm-2mv, trypsin, and pbs in water bath, wash with pbs (10ml each flask, aspirate the pbs. Note 1: to do in batches of 4. Note 2: it is empirical that the flasks get 5min. Any longer will kill the cells: take flasks out of incubator, check under microscope (for microscope dummy user: yes, place the flask under the lens). The cells should be floating together: smack the flask forcefully 4 times, add 7ml egm-2mv, to deactivate trypsin.

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