Efstathios Tsoukalos was born in Missolonghi and graduated from the Palmaic School of Missolonghi in 1964. In the same year, he entered the Medical School of Athens from which he graduated in 1970. He attended his military service for two years (1971-1973) as a hoplite doctor and then his tenure as an agricultural doctor (1973-1974) at the Agricultural Clinic of Stamna, Missolonghi. From 1974 to 1978 he served successively in the G.K.N.A. hospitals (later G. GENNIMATAS Hospital), ELPIS and ALEXANDRA hospitals at the First University Hospital Obstetrics-Gynecology Clinic of the Medical School of Athens, from where he received the specialty of obstetrician-gynecologist. From 1979 to 1986 he remained as a specialist assistant at the First University Clinic of N. ALEXANDRA, where he also prepared his doctoral dissertation and was awarded a Doctorate from the Medical School of the University of Athens in 1984.

He then worked in the private sector and in 1993 he was hired as a curator at ELENA VENIZELOU Maternity Hospital, remaining as Director until 2013. Since then, he has been working as a freelancer in the private sector. Beyond and in parallel with his scientific and professional career, he has developed intense and long-term trade union activity.