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Tebubio's blog - Acting and reacting in life sciences and biotechnologies
  • Home
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    • ADME-Tox
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    • Cell Biology and Signalling
    • Cell Sourcing – Cell Culture Technologies
    • Drug Discovery
    • Gene Expression – Molecular Biology
    • Stem Cells
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Supplying Discovery Tools

Fluorescence microscopy: how to improve your Live Cell Imaging

09/03/2017 by Frédéric Samazan No Comments

For more than 2 years now, the Silicon Rhodamine-like (SiR) technology has allowed the live cell imaging field with fluorescence microscopy to evolve significantly.

Fluorescent SiR-probes have appeared as the best alternative tools for studying Actin (SiR-actin), Microtubules (SiR-Tubulin), DNA (SiR-DNA) and now lysosome (SiR-Lysosome) for live cell imaging. Who better to show this? Well, here’s how other researchers have been using them to get optimal results.

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Cell Biology and Signalling

SiR fluorogenic probes: multicolour live-cell Imaging of Actin, Tubulin, DNA, and Lysosomes

05/09/2016 by Ali El Baya, PhD No Comments

The Silicon Rhodamine-like (SiR) technology has significantly contributed to the recent development of DNA and cytoskeletal analysis by live cell imaging.

Spectrum SiR 655. Spirochrome - tebu-bio.In 2014, two new Silicon Rhodamine-like (SiR) fluorescent probes were released for studying actin & tubulin by live cell imaging. SiR-Actin and SiR-Tubulin are fluorescent probes compatible with most microscopes (including super-resolution settings) that directly stain actin & tubulin without the need to transfect cells with vectors expressing fluorescently labeled Actin or Tubulin. The two original dyes were successfully followed by a new SiR-DNA probe in order to visualize DNA in living cells.

The existing SiR stains have a λabs of 652 nm and a λem of 674 nm to be used with the Cy5 filter (Fig 1).

However, the continuously growing number of researchers using these stains asked us whether stains with different biophysical properties would be made available. In other words, they were asking “is there another colour to allow for double staining e.g. of Actin and Tubulin in living cells?”

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Cell Biology and Signalling

Verapamil can enhance live cell staining of Actin & Tubulin with SiR-dyes

12/02/2015 by Ali El Baya, PhD No Comments

SiR-actin and SiR-tubulin kits now contain Verapamil

Fig 3 c -

STED image (raw data) : axons of rat primary hippocampal neurons stained with SiR-actin at 16 days in vitro

Recently, we were pleased to launch highly innovative tools to stain actin and tubulin in living cells without the need to transfect cells with vectors coding for GFP- or RFP tagged proteins which bind to filamentous cytoskeletal structures. This makes the SiR stains produced by Spirochrome the only tools available on the market which allow direct live cell imaging of actin and tubulin. I introduced you to this technology, as well as the benefits of SiR-actin and SiR-tubulin, in a recent post 2 new Actin and Tubulin live-cell imaging stains – without transfection.

Quite a number of cell types have already been successfully stained with SiR dyes, e.g. HeLa cells, Vero cells, BHK cells and a lot more cell lines, as well as primary cells such as HUVECs cells, dermal fibroblasts, and hippocampal neurons.

However, it turned out that some cell types, especially cell lines, do not sufficiently take up the dye. In these cases, the addition of Verapamil usually increases the uptake efficiency significantly and results in satisfying staining.

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Cell Biology and Signalling

2 new Actin and Tubulin live cell imaging stains – without transfection!

30/10/2014 by Ali El Baya, PhD 10 Comments

Cytoskeletal live cell imaging is extremely powerful when investigating cellular processes such as cytokinesis, motility and organelle transport and organization. The current experimental procedures remain nevertheless cumbersome and long. This post demonstrates how cell permeable, transfection free, Tubulin and Actin red fluorescent dyes help Cell biologists in analysing cytoskeleton dynamics in living cells.

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