What is the definitive moment in the play Trifles by Susan Glaspell?

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The clear definitive moment in the play comes when both Mrs. Hale and Mrs. Peters realise that the dead bird they have found is the one piece of evidence that definitely links Minnie Wright to the murder of her husband. Although Mrs. Peters pretends that this dead bird is actually unimportant, at the same time as she tries to state this opinion, the County Attorney is heard saying the following:

If there was some definite thing. Something to show--something to make a story about--a thing that would connect up with this strange way of doing it.

What both Mrs. Hale and Mrs. Peters realise at that precise moment is that they have discovered precisely that "definite thing" that the County Attorney is looking for, and it is the dead bird that would allow him to "connect up" and "make a story" about the death of John Wright, allowing Minnie to be convicted with his murder. This part of the play is so climactic precisely because the women realise that what they do or do not do will have profound ramifications for Minnie and result in her imprisonment or her freedom. As she has actually been imprisoned for all of her married life, their actions are designed to give her freedom, as in an act of female solidarity, the two women act to hide the dead bird.