Many critics view the themes of gender roles and sexuality in the play as a battle for power between men and women. Numerous commentators believe that this struggle ultimately concludes with the male characters prevailing. Angelo's attempt to assault Isabella and the Duke's manipulation involving the bed-trick are frequently analyzed as methods through which male characters reclaim control over women they see as disruptive or sexually assertive. Some critics have observed that the play's portrayal of fear towards women was typical of the Renaissance period, during which Shakespeare wrote. Furthermore, later works by other authors expressed even more intense distrust of women.

Scholars hold differing views on the marriages that conclude the play. Some suggest that the Duke's orchestration of engagements and weddings reaffirms male authority over women. In contrast, others see these marriages as a way to restore equilibrium between the genders.

Several critics explore the theme of female chastity within the play. Some assert that Shakespeare both acknowledges and critiques the double standard surrounding premarital sex, where a woman—expected to uphold chastity but suspected of being a seductress—was traditionally blamed for leading her lover astray. Critics also propose that Measure for Measure distinguishes between the rigid, celibate chastity of the novice Isabella and the warm, emotional chastity of the devoted Mariana.

The themes of justice and mercy are considered fundamental in Measure for Measure. Scholars frequently compare these concepts, describing "justice" as the strict application of the law, while "mercy" involves a more compassionate and flexible interpretation. Traditionally, Angelo (and to some degree Isabella) is portrayed as a rigid enforcer of justice, whereas Duke Vincentio is seen as someone who tempers justice with mercy. Critics have noted the Duke's admission in Act I of his own leniency in enforcing Vienna's laws, as well as his concern that Angelo might be excessively strict. Many conclude that the play seeks to find a middle ground between these two positions.

The question of whether such a balance is achieved has led to much debate. Some scholars argue that Angelo cannot judge fairly until he has personally sinned. Others believe that personal experiences should not influence the execution of justice. These issues reach a climax in Act V when Vincentio initially imposes harsh punishments on Angelo and Lucio, only to later pardon them. The fairness and wisdom of the Duke's justice, softened by mercy, continue to be debated. To achieve his goals, Duke Vincentio employs the bed-trick and misleads Isabella into believing her brother is dead. This has prompted many critics to question the morality of achieving justice through deception and falsehoods.

Determining whether Measure for Measure lacks coherence is a complex issue. The play has been criticized for its seemingly inconsistent characters, unresolved themes, and ambiguous genre. As a result, many scholars scrutinize the play's structure to assess its overall unity. Two structural elements, in particular, have attracted attention. Firstly, the play consists of scenes that focus on either action or dialogue. Action scenes drive the plot forward, while dialogue scenes give the audience an opportunity to reflect on the significance of the events.

Secondly, in response to assertions that Measure for Measure does not fit neatly into a single genre, several critics maintain that the play's unity is found in its two-part structure. In the tragic first half, Claudio is sentenced to death, Isabella is threatened with rape, and Angelo strays from his rigid moral principles. In the comedic second half, Duke Vincentio steps in to judge Angelo, save Claudio from execution, protect Isabella from rape, and orchestrate several marriages. Some commentators, therefore, suggest that the play achieves unity as a tragicomedy.