Dear friend J. Ramirez-Franco

Finding a specific commercially available tdTomato-nanobooster for clearing experiments can be challenging, as the availability of conjugated antibodies or nanobodies can vary. However, there are alternative strategies you can consider for enhancing tdTomato signal in 3D microscopy. Here are a few options:

1. Amplification techniques: Instead of using a tdTomato-specific nanobooster, you can employ amplification methods to enhance the fluorescence signal. One commonly used technique is the tyramide signal amplification (TSA) system, which utilizes horseradish peroxidase (HRP) and fluorophore-conjugated tyramide. This approach can increase the fluorescence signal and enable better detection of tdTomato.

2. Direct labeling: If a tdTomato-specific nanobooster is not readily available, you can directly label the tdTomato protein using fluorescent dyes or nanoparticles. This can be achieved by employing chemical conjugation methods or genetic engineering techniques to introduce fluorescent tags directly onto the tdTomato protein.

3. Genetic alternatives: Instead of using tdTomato, you can consider using other red fluorescent proteins (RFPs) that are commercially available with specific nanoboosters. For example, mCherry is another red fluorescent protein widely used in imaging studies, and there are conjugated antibodies or nanobodies available for amplifying mCherry signal.

It is important to note that the choice of the appropriate strategy will depend on the specific requirements of your experiment and the compatibility with your clearing technique. Additionally, it is advisable to consult the latest research articles, protocols, or supplier catalogs for up-to-date information on the availability of tdTomato-specific nanoboosters or alternative methods for enhancing tdTomato signal in 3D microscopy.

Dear Kaushik Shandilya ,

Thank you so much for your answer.

Indeed, after searching a little bit more I found a commercially available anti-RFP that apparently recognizes tdTomato

https://nano-tag.com/product/fluotag-x4-anti-rfp/

Thanks though for providing alternatives,

Cheers,

J

Dear colleague

here a commercial link which potentially help you:

https://www.anticorps-enligne.fr/antibody/6254170/anti-tdTomato+Fluorescent+Protein+tdTomato+antibody/

Then you can contact my colleagues of the photonic unit of the Bordeaux Imaging Center at: https://www.bic.u-bordeaux.fr/team/

Best regards

Philippe

Dear Philippe Le Coustumer ,

Thanks for your answer.

Unfortunately, I am looking for a nanobody (aprox 15KDa), not an antibody (aprox 150KDa), so the alternative you are providing is not valid for me in this situation.

Thanks for your help and your time in any case.

Best regards,

J