Label-free imaging of cell attachment with photonic crystal enhanced microscopy

• Published in: Analyst (London) 2011-09, Vol.136 (18), p.3608-3615
• Main Authors: LIDSTONE, Erich A, CHAUDHERY, Vikram, KOHL, Anja, CHAN, Vincent, WOLF-JENSEN, Tor, SCHOOK, Lawrence B, BASHIR, Rashid, CUNNINGHAM, Brian T
• Format: Article
• Language: English
• Subjects: Ageing, cell death; Analytical chemistry; Animals; Apoptosis; Attachment; Biological and medical sciences; Biosensing Techniques - methods; Biosensors; Biotechnology; Cancer; Cell Adhesion - physiology; Cell Differentiation; Cell physiology; Cells, Cultured; Chemistry; Density; Exact sciences and technology; Extracellular Matrix - metabolism; Fundamental and applied biological sciences. Psychology; General, instrumentation; Humans; Integrins - metabolism; Methods. Procedures. Technologies; Microscopy; Microscopy, Phase-Contrast; Molecular and cellular biology; Monitors; Photonic crystals; Photons; Rats; Rats, Sprague-Dawley; Stem cells; Swine; Various methods and equipments
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/c1an15171a

We introduce photonic crystal enhanced microscopy (PCEM) as a label-free biosensor imaging technique capable of measuring cell surface attachment and attachment modulation. The approach uses a photonic crystal optical resonator surface incorporated into conventional microplate wells and a microscope-based detection instrument that measures shifts in the resonant coupling conditions caused by localized changes in dielectric permittivity at the cell-sensor interface. Four model systems are demonstrated for studying cancer cells, primary cardiac muscle cells, and stem cells. First, HepG2/C3 hepatic carcinoma cells were cultured and observed via PCEM in order to characterize cell adhesion in the context of growth and locomotion. Second, Panc-1 pancreatic cancer cells were used to verify that cell attachment density decreases in response to staurosporine, a drug that induces apoptosis. Third, we used PCEM to confirm the influence of integrin-mediated signaling on primary neonatal cardiomyocyte growth and development. Rounded cardiomyocytes consistently showed decreased cell attachment density as recorded via PCEM, while spreading cells exhibited greater attachment strength as well as increased contractility. Finally, PCEM was used to monitor the morphological changes and extracellular matrix remodeling of porcine adipose-derived stem cells subjected to a forced differentiation protocol. Each of these experiments yielded information regarding cell attachment density without the use of potentially cytotoxic labels, enabling study of the same cells for up to several days.