A strong tradition in educational research has involved several widely-used instruments assessing student or teacher perceptions of characteristics of actual or preferred classroom psychosocial environment. Existing instruments, however, are unsuitable for one of the most important settings in science teaching, namely, the science laboratory class. Consequently, the present research aimed to develop and validate a new instrument, the Science Laboratory Environment Inventory (SLEI), which is specifically suited to science laboratory environments at the higher education level. The SLEI assesses students' or teachers' perceptions of five dimensions of actual or preferred classroom environment, namely, Student Cohesiveness, Open-Endedness, Integration, Rule Clarity, and Material Environment. A distinctive feature of the design of the study was that the instrument was field tested and validated cross-nationally in six different countries, namely, the USA, Canada, Australia, England, Israel, and Nigeria. The total sample consisted of 1720 students in 71 university laboratory classes. Various item and factor analyses guided the evolution of a refined version, and attested to each SLEI scale's internal consistency reliability, discriminant validity, factorial validity, predictive validity (i.e., ability to predict student outcomes), and ability to differentiate between the perceptions of students in different classes. An important finding was that the SLEI is equally valid for use in its actual and preferred versions, and for the individual or the class mean as the unit of analysis. As well, separate within-country analyses confh-med the validity and usefulness of the SLEI in each of the six countries involved in the field testing. Overall, the study attested to the general advantages of employing cross-national designs in learning environment research.