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TIRF

Using Evanescent Waves for High Resolution Membrane Fluorescence Imaging

Fig. 1 A/B: (A) Brightfield photomicrograph of human myotubes. (B) The same cells observed with the SRIC filter. The dark grey outline represents the footprint of the cell on the glass coverslip. This focal plane was selected to perform time lapse imaging of Ca2+ influx.
Fig. 1 A/B: (A) Brightfield photomicrograph of human myotubes. (B) The same cells observed with the ... more
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Changes in the intracellular Ca2+ concentration ([Ca2+]i) accompany many diverse biological phenomena from neuronal excitability and muscle contraction to activation of transcription and cell death. Thanks to the introduction of fluorescent intracellular Ca2+ dyes our understanding of Ca2+ signalling has greatly advanced; it is established that depending on the signalling molecule and cell type, the global increase in cytoplasmic Ca2+ is mediated by the mobilisation of Ca2+ from intracellular stores and by the opening channels on the plasma membrane, or by a combination of both mechanisms. In this application note, we use TIRF to monitor intracellular free calcium ion concentration [Ca2+]i on or very close to the plasma membrane  of muscle cells including human myotubes.

 

Authors:
Susan Treves
Francesco Zorzato

Keywords: biomedical inverted research microscope Brightfield Fluorescence Imaging Fluorescence Microscopy Microscopy Nikon Nikon Instruments Perfect Focus System PFS SRIC TIRF

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