Background: Facial expression, gesture, and posture play an important role in perceiving emotions during communication. In virtual communication platforms, users have devised and learned to use a variety of expressive emojis along with text messages to express certain emotions. The effect of emojis on human psychology and associated autonomic responses is still vague. Methods: A total of 100 healthy individuals (50 males and 50 females) aged between 18-40 years were recruited. Electrodermal activity (skin conductance level (SCL) and skin conductance response (SCR) amplitude) and Respiratory Sinus Arrhythmia (RSA) were assessed during the Emotional Stroop Task (EST). EST having expressive emojis superimposed with the congruent and incongruent words was used. Result: Mean SCL and SCR amplitude was significantly increased during EST in incongruent and congruent blocks as compared to neutral block (p < 0.001). RSA was significantly decreased in incongruent and congruent blocks as compared to neutral blocks (p < 0.001). We found an increased sympathetic activity and parasympathetic withdrawal while performing the task. Conclusion: The results of this study suggested that emojis are adequate stimuli to elicit autonomic responses and change both sympathetic (EDA) as well as parasympathetic responses (RSA). Males and females showed similar autonomic arousal for emoji but the baseline emotional status was different for both genders. Keywords: Autonomic nervous system, Electrodermal response, Respiratory sinus arrhythmia, Emotions, emotional Stroop task.