Label free imaging of cell-substrate contacts by holographic total internal reflection microscopy.
The study of cell adhesion contacts is pivotal to understand cell mechanics and interaction at substrates or chemical and physical stimuli. We designed and built a HoloTIR microscope for label-free quantitative phase imaging of total internal reflection. Here we show for the first time that HoloTIR is a good choice for label-free study of focal contacts and of cell/substrate interaction as its sensitivity is enhanced in comparison with standard TIR microscopy. Finally, the simplicity of implementation and relative low cost, due to the requirement of less optical components, make HoloTIR a reasonable alternative, or even an addition, to TIRF microscopy for mapping cell/substratum topography. As a proof of concept, we studied the formation of focal contacts of fibroblasts on three substrates with different levels of affinity for cell adhesion.
CNR - ISASI Institute of Applied Sciences and Intelligent Systems, Via Campi Flegrei 34, 80078, Pozzuoli, Napoli, Italy.|Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale - DICMAPI, University of Naples Federico II, Piazzale Tecchio 80, 80100, Napoli, Italy.
Biagio
B
Mandracchia
CNR - ISASI Institute of Applied Sciences and Intelligent Systems, Via Campi Flegrei 34, 80078, Pozzuoli, Napoli, Italy.
Oriella
O
Gennari
CNR - ISASI Institute of Applied Sciences and Intelligent Systems, Via Campi Flegrei 34, 80078, Pozzuoli, Napoli, Italy.
Valentina
V
Marchesano
CNR - ISASI Institute of Applied Sciences and Intelligent Systems, Via Campi Flegrei 34, 80078, Pozzuoli, Napoli, Italy.
Melania
M
Paturzo
CNR - ISASI Institute of Applied Sciences and Intelligent Systems, Via Campi Flegrei 34, 80078, Pozzuoli, Napoli, Italy.
Pietro
P
Ferraro
D000818:Animals; D002448:Cell Adhesion; D005347:Fibroblasts / Q000166:cytology*; D006696:Holography; D051379:Mice; D008853:Microscopy / Q000379:methods*; D041681:NIH 3T3 Cells
Cell Membrane [A11.284.149]; Cell Shape [G04.320]; Interference [E05.595.513]; Microscopy; Microscopy [E05.595]