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Handshake frequency in the NICU decreased during the handshake-free zone trial.The impact of the handshake-free zone varied with gender and profession.Male professionals shook hands with families more commonly than female professionals.Most families and health care providers supported the idea of a handshake-free zone.The handshake represents a social custom with special importance in health care settings. However, handshakes can transmit disease and compliance with hand hygiene protocols averages <50%. We hypothesized that a handshake-free zone (HFZ) could be established within our neonatal intensive care unit (NICU) and would be well-received by patient families and their health care providers (HCPs).We established an HFZ and conducted a prospective cohort study in the NICU at 2 UCLA Medical Centers. Data collection tools included questionnaires for NICU families and their HCPs.Handshake greetings occurred more frequently before than during the HFZ, as reported by HCPs (P = .0002) and patient families (P = .05). Before the HFZ, physicians were more likely than nurses to shake hands with patient families (P = .001), and believe the handshake was extremely important (P = .002); during the HFZ physicians' behaviors and attitudes shifted toward those of the nurses. All patient families and 66% of HCPs believed the NICU should consider establishing an HFZ.The HFZ decreased the frequency of handshakes within the NICU. The influence of the HFZ on HCP behavior and attitudes varied with gender and profession. Patient families and most HCPs supported the implementation of an HFZ.