The　behaviors　of　cells　cultured　on　patterned　substrates　vary　with　the　material　stiffness,　the　geometry　and　the　biochemical　properties　of　the　pattern.　By　using　a　reversed　cell　imprinting　(RCI)　technique,　together　with　phase　contrast　microscopy,　scanning　electron　microscopy　(SEM)　and　atomic　force　microscopy　(AFM),　we　have　exploited　reversed　side　cellular　morphology　on　patterned　microgrooves　of　different　geometries　with　or　without　surface　coating　of　adhesion　molecules.　We　have　shown　a　close　correlation　between　the　effect　of　contact　guidance　and　penetration　of　cellular　membrane.　Without　surface　coating,　roughly　80%　of　HeLa　cells　were　aligned　along　the　groove　direction　regardless　of　the　groove　spacing.　When　the　microgrooves　were　coated　with　fibronectin,　the　area　of　cell　spreading　was　increased　but　the　percentage　of　aligned　cells　was　significantly　decreased.　In　both　cases,　the　deformation　of　cell　membrane　at　the　cell-pattern　interfaces　could　be　measured.　We　found　that　the　local　penetration　of　the　cellular　membrane　into　the　grooves　was　correlated　to　the　cellular　alignment　for　both　HeLa　and　NIH　313　cells,　and　that　such　a　correlation　was　cell-type　dependent.　(C)　2012　Elsevier　B.V.　All　rights　reserved.