Transparency of samples
FLO-400: Safe, fast and easy to use search device.
Transparent embryos, the technique of transparency
Although different protocols have emerged one after the other in recent years, the complexity of some has made them extremely complicated to use under laboratory conditions, particularly through the use of harmful reagents.
We have studied the use of 6 different protocols to adapt 3 of them to our Flo400 PLC.
The fastest, the 3DISCO:
This protocol is the shortest since the current total time to perform it on our PLC is 14 hours. After a passage of tetrahydrofuran at increasing concentration in the samples, dichloromethane and di-benzyl ether stabilize the samples until fully transparent samples are obtained. This Protocol makes it possible to preserve the sizes of the embryos but also the fluorescent markings obtained after Immunohistochemistry. Be careful, however, Tetrahydrofuran is a highly toxic reagent and requires handling under a hood. In addition, this compound destroys some plastics such as polypropylene, so you will need to use polystyrene tankers.
ClearT2, to maintain proportions:
This protocol is very effective for the clarification of samples containing fluorescent mutated cells. In just 16 hours, your sample becomes completely transparent and retains full fluorescence expression in the desired cells. In addition, this protocol keeps the dimensions of your sample completely intact. It also preserves fluorescent immunomarkings. However, it uses Formamide, which is a harmful compound and therefore requires to be used in fume hoods. In addition, the transparency of the sample is stable for only one or two days before becoming opaque again (however, another passage in the ClearT2 protocols makes it transparent again.)
The SeeDB, the least harmful and most economical:
This protocol, a little longer than the other 2 (24 hours with the Flo 400, 96 in manual mode) is the most economical and least harmful of the three. It consists of incorporating a fructose solution with increasing concentration gradually into the sample. The solutions are composed solely of fructose and α-thioglycerol (irritant product) which take the place of the fatty acids present and make the embryo transparent. With this protocol, transparency and fluorescence are maintained for more than 3 weeks if the samples are obscured. Currently, Flogentec has the record for the length of time it has been clarified by this protocol.
These three protocols (SeeDB, 3DISCO and ClearT) have been adapted by our team to allow you to clarify your samples easily and fully automatically. The performance offered by the Flo 400 allows you to clarify more than 50 samples in a single protocol by significantly reducing the amount of reagents required through its recycled flow system.
The Flo400 PLC can also perform the techniques:
Contact our advisors
Research in biology is constantly diversifying and becoming more complex with the emergence of new technologies and the accumulation of knowledge. Thus, different fields of applications and experiments in biology are emerging and new high throughput biology techniques are emerging.
In situ hybridization, immunohistochemistry and fluorescence in situ hybridization are tagging methods that provide a clear understanding of the expression of each gene in a spatial and temporal context within a given sample. This allows a characterization of the proteome or genome of the organism studied.
Flogentec thus offers biological automatons that can have several different applications depending on your research field.
The Flo400 is a safe, fast and easy-to-use research device that allows 3D samples to be analyzed using the in situ hybridization, transparency of samples and immunohistochemistry methods. Thanks to this equipment, it is possible to directly visualize the location and activation of genes of the cells concerned. The means used for this localization is in situ hybridization (HIS) of RNA probes (transcriptome analysis). It should be noted that the automaton also allows proteome analysis by immunohistochemistry, using antibodies instead of RNA probes.
Flo400 Wholemount PLC Wholemount: This ISH/IHC PLC for in toto samples is specialized for biological research.
Flo400 Slide PLC: This ISH/IHC PLC for slide samples is designed for biological research and diagnosis.
Flo400 Combo PLC: This ISH/IHC PLC for all types of samples is specialized for biological research.