RESUMO
Fluorescence correlation spectroscopy (FCS) is a popular technique, complementary to cell imaging for the investigation of dynamic processes in living cells. Based on fluorescence, this single molecule method suffers from artifacts originating from the poor fluorophore photophysics: photobleaching, blinking, and saturation. To circumvent these limitations we present here a new correlation method called photothermal absorption correlation spectroscopy (PhACS) which relies on the absorption properties of tiny nano-objects. PhACS is based on the photothermal heterodyne detection technique and measures akin FCS, the time correlation function of the detected signals. Application of this technique to the precise determination of the hydrodynamic sizes of different functionalized gold nanoparticles are presented, highlighting the potential of this method.
Assuntos
Processos Fotoquímicos , Análise Espectral/métodos , Absorção , Difusão , Ouro/química , Luz , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Nanopartículas Metálicas/química , Movimento (Física) , Temperatura , Fatores de TempoRESUMO
The far-field optical imaging of mitochondria of live cells without the use of any label is demonstrated. It uses a highly sensitive photothermal method and has a resolution comparable to confocal fluorescence setups. The morphological states of mitochondria were followed under different physiological treatments, and the role of cytochrome c was ruled out as the main origin of the photothermal signals. This label free optical method provides a high contrast imaging of live mitochondria and should find many applications in biosciences.
RESUMO
Tracking individual nano-objects in live cells during arbitrary long times is a ubiquitous need in modern biology. We present here a method for tracking individual 5-nm gold nanoparticles on live cells. It relies on the photothermal effect and the detection of the Laser Induced Scattering around a NanoAbsorber (LISNA). The key point for recording trajectories at video rate is the use of a triangulation procedure. The effectiveness of the method is tested against single fluorescent molecule tracking in live COS7 cells on subsecond timescales. We further demonstrate recordings for several minutes of AMPA receptors trajectories on the plasma membrane of live neurons. Single Nanoparticle Photothermal Tracking has the unique potential to record arbitrary long trajectory of membrane proteins using nonfluorescent nanometer-sized labels.
Assuntos
Ouro/química , Nanopartículas Metálicas , Animais , Transporte Biológico , Células COS , Carbocianinas , Membrana Celular/metabolismo , Células Cultivadas , Chlorocebus aethiops , Corantes Fluorescentes , Lasers , Luz , Microscopia de Fluorescência , Neurônios/metabolismo , Receptores de Glutamato/metabolismo , Espalhamento de Radiação , TemperaturaRESUMO
The dynamics of heterogeneities in a shear thickening, concentrated colloidal suspension is investigated through speckle visibility spectroscopy, a dynamic light scattering technique recently introduced [P. K. Dixon and D. J. Durian, Phys. Rev. Lett. 90, 184302 (2003)]. Formation of shear-induced heterogeneities is observed in the jamming regime, and their relaxation after shear cessation is monitored as a function of the applied shear stress. The relaxation time of these heterogeneities increases when a higher stress is applied.