. Author manuscript; available in PMC: 2019 May 1.

Published in final edited form as: Curr Protoc Toxicol. 2018 May;76(1):e46. doi: 10.1002/cptx.46

Table 1.

Commonly Encountered Problems and Solutions.

Protocol	Problem	Possible cause(s)	Solution (current experiment)	Solution (future experiments)
Isolation	No platelet pellet present after wash step	Platelets were activated	--	Ensure PGI₂ is added during wash/isolation steps
Isolation	No platelet pellet present after wash step	Platelets were activated	--	Ensure platelets do not contact cold materials
Isolation	Platelets released from pellet in chunks	Platelets were activated	Let the platelet pellet rest for 10 minutes after spin	Ensure PGI₂ is added to PRP and wash buffer
Aggregation	No aggregation	Reagent bad	Test/use fresh reagent	Do not refreeze agonist once thawed
Aggregation	No aggregation	Donor insensitive to agonist	Use other agonists	-
Spreading	No platelets on coverslips	Coverslip flipped	-	Care in the future to mount the correct side of the coverslip
CSE Generation	Cigarette smoke not bubbling into media	Clogged tubing	Replace tubing	Regularly check and replace tubing
CSE Generation	Cigarette smoke not bubbling into media	Leak in tubing	Check tubing for leaks, replace tubing	Regularly check and replace tubing