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Aseptic techniques are essential in preventing infections in neonatal care, a field where even minor lapses can have significant consequences. Evaluating the knowledge of staff nurses, who are on the front line of neonatal care, are crucial to maintaining high standards of patient safety and care quality. Objectives of the study : 1. To assess the knowledge of staff nurses regarding aseptic techniques in neonatal care. 2. To identify areas of strength and areas need for improvement in staff nurse knowledge regarding aseptic techniques in neonatal care. 3. To know the relationship between demographic factors (e.g., gender, age, educational background, experience) and knowledge levels. Methods : Methods Design: A cross-sectional descriptive study design. Setting: Selected hospitals in Bangalore, India. Participants: 85% female and 15% male nurses participated in the study, with a mean age of 29.5 years. The educational background of the participants included 50% with a Diploma in Nursing, 40% with a Bachelor's degree, and 10% with a Master's degree. Experience varied, with 40% of nurses having 1-3 years of experience, and 35% having received specialized training in neonatal care. Data Collection: A structured questionnaire was used to assess the knowledge of aseptic techniques, covering areas such as hand hygiene, sterilization procedures, and the use of personal protective equipment. The questionnaire was validated for content and reliability. Data Analysis: Knowledge scores were calculated, with mean scores, SD and percentages used to categorize the levels of knowledge. Statistical analysis, including Chi-square tests and ANOVA, was conducted to find out the influence of demographic factors on knowledge scores. Results Knowledge Scores: The mean knowledge score was 75.2% (SD = 10.4), with a range from 50% to 90%. Distribution of scores showed that 20% of nurses were in the "Excellent" category (85-100%), 50% in the "Good" category (70-84%), 25% in the "Fair" category (50-69%), and 5% in the "Poor" category (below 50%). ? Strong Knowledge Areas: Nurses demonstrated strong knowledge in hand hygiene (90% correct) and the use of personal protective equipment (85% correct). ? Areas Needing Improvement: Lower scores were observed in sterilization procedures (60% correct), handling of sterile equipment (65% correct), and infection control measures (70% correct), indicating areas where further training is required. ? Influencing Factors: Higher knowledge scores were significantly associated with higher educational qualifications (Bachelor's or Master's degree), more than 3 years of experience, and specialized training in neonatal care (p < 0.05).