In this study , we explore the application of our laser - guided cell micropatterning ( LGCM ) system 15 in combination with surface patterning methods 16 to investigate stem cell differentiation at the single - cell level in a cardiomyocyte microculturing environment . In previous studies , we determined the effect of cell - cell contact on MSC cardiogenic differentiation by creating a microenvironment with only one MSC and one cardiomyocyte using the LGCM system . 17 In the study reported here , we first constructed a cardiomyocyte culture model with the controlled alignment of cardiomyocyte constructs , and then utilized LGCM to trap and deposit individual MSCs into the constructed model . Next , we evaluated cell differentiation at the single - cell level through single - cell RT - qPCR and patch - clamp assays . Consequently , we report ( i ) the construction of a laser - patterned , biochip - based , stem cell - cardiomyocyte coculture model with controlled cell alignment ; and ( ii ) single - cell - level data on stem cell cardiogenic differentiation under an in vivo - like cardiomyocyte alignment conditions .