Her early series Being and Having (1991) and Portraits (1993—1997) depicted her friends in the lesbian and gay community in Los Angeles, mixing traditional portrait photography with less traditional subjects. In 1994, with Freeways, she made images of the highways around her Los Angeles home that exclude people and automobiles from the compositions. She continued this exploration of the contemporary landscape three years later with Mini-malls. In the Domestic (1999) series, she returned to the lesbian community for her subjects, photographing couples in their homes. Polaroids (2000) documents the performance work of Ron Athey in large-scale prints. Traveling outside the Los Angeles area to find new landscapes and cityscapes to photograph, she photographed the New York financial district, once again emptied of people, in Wall Street and scenes of Minnesota in Icehouses and Skyways (all 2001). A 2003 series of portraits entitled Surfers focuses on the California surfing subculture. For her series Children (2004), Opie returned to the studio and to her signature highly focused portraits, this time of children set against bright solid backdrops. In 2004 the artist extended her study of unpeopled urban sites initiated in 1997, now grouped under the title American Cities, with black-and-white photographs of Chicago. For the series In and Around the Home (2004–05), Opie turned to her home and community as a microcosm for the larger political and social conflicts surrounding the presidential election. In another group of studio portraits from 2013, Opie evoked seventeenth-century portraiture, presenting her subjects in allegorical poses in front of black backgrounds, which remove the individuals from any sense of time or place.

Opie has had solo shows at the Museum of Contemporary Art Los Angeles (1997); Saint Louis Art Museum (2000); The Photographers’ Gallery, London (2000); Walker Art Center, Minneapolis (2002); Aldrich Contemporary Art Museum, Ridgefield, Connecticut (2006); the Museum of Contemporary Art, Chicago (2006); the Solomon R. Guggenheim Museum, New York (2008); the Los Angeles County Museum of Art (2010); Portland Art Museum (2010–11); Institute of Contemporary Art, Boston (2011); and the Wexner Center for the Arts, Ohio State University, Columbus (2015); among other venues. Her work has appeared in the Whitney Biennial (1995 and 2004), the Melbourne International Biennial (1999), and SITE Santa Fe Biennial (2006), as well as in group exhibitions at Centre Georges Pompidou, Paris (1995); the Solomon R. Guggenheim Museum (1997, 2007); Guggenheim Museum Bilbao (2003); Brooklyn Museum (2007, 2011); The Museum of Modern Art, New York (2009); J. Paul Getty Museum, Los Angeles (2010); Museo Nacional Centro de Arte Reina Sofía, Madrid (2010); National Portrait Gallery, Washington, D.C. (2010); the Museum of Contemporary Art Los Angeles (2010, 2011, 2014–15); and the Whitney Museum of American Art, New York (2013, 2015). She has taught at Yale University and at the University of California, Los Angeles. She lives and works in Los Angeles.