Overview

  • Product name
    CyGEL™
  • Tested applications
    Suitable for: FM, ICC/IFmore details
  • General notes

    CyGEL™ is a novel thermoreversible hydrogel which is liquid when cold and a gel when warmed at room temperature (22°C +/- 1°C). CyGEL™ is compatible with live cells and can be used to immobilize non-adherent cells by simple warming, and conversely, allowing their recovery by simple cooling.
    CyGEL™ has many applications in imaging and cell-based screening.
    Key features of CyGEL™ include:

    1. Convenient immobilization of non-adherent cells.
    2. Controllable and rapid transition from liquid to gel, which allows quick preparation of live cells for gel mounting.
    3. Optically clear with low autofluorescence.
    4. Compatible with GFP and other fluorescent probes such as DRAQ5™ (ab108410).


    CyGEL™ is recommended for mounting live de-adherent/ non-adherent cells for microscopy, or mounting live C. elegans and parasites (e.g. leishmania, trypanosomes), which can be recovered afterwards. CyGEL™ is provided as concentrated non-buffered form. Add the concentrated PBS supplement (provided) to prime CyGEL™ prior use.
    CyGEL™ is supplied as 8 vials, each containing 500µl, and 1 vial of 40xPBS.
    For imaging experiments longer than 2 hours, we recommend using CyGEL Sustain™ (ab109205).

Properties

Applications

Our Abpromise guarantee covers the use of ab109204 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
FM Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Please refer to the Protocol Booklet for detailed information about the use of CyGEL.

Images

  • This ilustration demonstrates the general procedure for using CyGEL™ with live cells prior analysis by fluorescence microscopy.

Protocols

References

ab109204 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

excellent method

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This is a good method to perform single cell analysis and afterwards to cultivate the cells again or to make additional testings.
It works well and reliably.
The instructions are easy to understand and easy to implement.
We have used it for immunofluorescence analysis.

Abcam user community

Verified customer

Submitted Oct 07 2015

Answer

Thank you for contacting us.



As suggested, there is no particular reason to use CyGEL Sustain over CyGEL (a PBS-based formulation) other than the ease of adding specific media.

We have limited data on the long-term imaging of embryos other than Danio which have been imaged for 24h.



The trick is to use a cover-glass bottomed petri-dish. Make a well with an 8-10 mm diameter silicone o-ring, smeared with silicone (aka vacuum) grease, to bed it down onto the cover glass surface. Surround the o-ring well with LMP agarose – this acts as a hydration buffer to stop the hydrogel from drying out. Fill the inner well with CyGEL and your embryo(s). Lid the petri-dish. You can now image through the cover glass.



For short-term use, i.e. injections, the embryos can be immobilized in a simple shallow well and the CyGEL melted and re-set as required to best position the organism for the injection. The organism(s) can then be gently recovered by melting by re-cooling.



The lab has done quite a bit of work with Drosophila embryos of a variety of ages and they seemed OK for around an hour at least – the limit of time worked on them. They are also aware of a user in Australia has imaged an embryo for close to 2h through 5-6 cell divisions.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us.

I am sorry you are not able to enter our Website, please do let us know if the problem persists. You will find attached the protocols for our CyGel products: ab109205 and ab109204.

These products should work in your application, otherwise please do not hesitate to contact us for assistance.

I hope this helps and if we can assist further, please do not hesitate to contact us.

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Answer

CyGEL is a Michelle structure and like most gels will not permit passive diffusion of macromolecules such and large proteins.

However, if the intention is to label cells with the tagged antibody in suspension and then wash and add them to CyGEL for mounting on a slide or in a well that is its ideal use. Once applied to the slide it will start to set. A coverslip can be added and then the CyGEL re-chilled to liquefy and allow the gel to spread under the coverslip.



This is the standard protocol for CyGEL.



Care should taken to avoid bubbles.



I hope that helps.Please let me know if you have any further questions.



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