A confocal.nl and Sandiabiotech collaboration

RCM imaging and FluorAbody staining, perfect combination for live cell tumor biomarkers detection

In vitro studies are part of the daily routine in cancer research. In this context, tumor biomarker detection and quantitation are key to develop new therapies and improve cancer diagnostics. However, live cell imaging  and precise quantitation can be challenging. On one hand, phototoxicity and photobleaching can alter cell behaviour and sample fluorescence. On the other hand, traditional labeling methods, like conjugated primary antibodies, can introduce crosslinking and have a variable binding site to fluorophore ratio, which makes quantition less precise.

Here we present, the perfect combination to overcome all those challenges and make live cell imaging and quantitation highly precise.

 

Sandia BioTech has developed a new labeling system, the FluorAbodies, to improve tumor biomarkers detection and quantitation. FluorAbodies are antibody analogues that comprise a single antigen-binding site (Fab) that is molecularly attached to a fluorescent protein. As a result, FluorAbodies are intrinsically fluorescent molecules,  monovalent and three times smaller than tradicional antibodies. Therefore, you save time and money, since there is no need for further labeling and the reduced size improves binding capabilities due to reduced steric hindrance and increases intracellular penetration. In addition, FluorAbodies present a unique ratio of 1:1 Fab to fluorescence protein, which makes them perfect for quantitation through stoichiometric measurements. 

 

FluorAbodies features:

  • Intrinsic fluorescence – no need of further labeling steps
  • Monovalent – no crosslinking
  • 3x smaller than conventional antibodies – better labeling capabilities
  • 1:1 ratio of binding site to fluorescence protein – precise quantitation
  • Available now: Anti-Her2, anti-EGFR, anti-EphA2, anti-ErbB3 and anti-ErbB4

 

Confocal.nl has built up the bright-i system, based on Re-scan Confocal Microscopy (RCM) technology. RCM uses a sCMOS as a detector resulting in 3x more sensitivity as compared to traditional confocal microscopes. The RCM re-scanning technology improves the resolution with 40%, resulting in 170nm resolution RAW images.  The RCM is the perfect imaging system to use in combination with FluorAbodies because it is highly sensitive, allowing the detection of small amounts of signal. 

 

RCM features:

  • High resolution (170nm lateral resolution in 488) – sharper images
  • High sensitivity  – with low laser power it detects weak signals
  • Low phototoxicity – Live cell imaging friendly
  • Easy to use

 

The combination of FluorAbodies and RCM bright-i system allows the precise imaging, detection and quantitation of very low amounts of probes, overcoming the challenges presented with traditional methods.

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