Bio-macromolecules,　such　as　proteins　and　nucleic　acids,　are　the　basic　materials　that　perform　fundamental　activities　required　for　life.　Their　structural　heterogeneities　and　dynamic　personalities　are　vital　to　understand　the　underlying　functional　mechanisms　of　bio-macromolecules.　With　the　rapid　development　of　advanced　technologies　such　as　single-molecule　technologies　and　cryo-electron　microscopy　(cryo-EM),　an　increasing　number　of　their　structural　details　and　mechanics　properties　at　molecular　level　have　significantly　raised　awareness　of　basic　life　processes.　In　this　review,　firstly　the　basic　principles　of　single-molecule　method　and　cryo-EM　are　summarized,　to　shine　a　light　on　the　development　in　these　fields.　Secondly,　recent　progress　driven　by　the　above　two　methods　are　underway　to　explore　the　dynamic　structures　and　functions　of　DNA,　antibody,　and　lipoprotein.　Finally,　an　outlook　is　provided　for　the　further　research　on　both　the　dynamic　structures　and　functions　of　bio-macromolecules,　through　single-molecule　method　and　cryo-EM　combining　with　molecular　dynamics　simulations.