Overview

  • Product name

    CyGEL Sustain™
  • Tested applications

    Suitable for: ICC/IF, FMmore details
  • General notes

    CyGEL Sustain™ is a novel thermoreversible hydrogel which is liquid when cold and a gel when warmed at room temperature (22°C +/- 1°C). CyGEL Sustain™ 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 Sustain™ has many applications in imaging and cell-based screening.
    Key features of CyGEL Sustain™ include:

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


    CyGEL Sustain™ is recommended for mounting live de-adherent/ non-adherent cells and Zebrafish embryos for microscopy. CyGEL Sustain™ is specially formulated to allow addition of RPMI and similar culture media, which makes it the best tool for extended imaging experiments, as cells will survive for several hours.
    CyGEL Sustain™ is supplied as 10 vials, each containing 500µl.
    For live C. elegans and parasites we recommend CyGEL™(ab109204).

Properties

Applications

Our Abpromise guarantee covers the use of ab109205 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
ICC/IF Use at an assay dependent concentration.

Please refer to the Protocol Booklet for detailed information about the use of CyGEL Sustain.

FM Use at an assay dependent concentration.

Images

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

Protocols

References

This product has been referenced in:

  • Fritzsche M  et al. Cytoskeletal actin dynamics shape a ramifying actin network underpinning immunological synapse formation. Sci Adv 3:e1603032 (2017). Read more (PubMed: 28691087) »
See 1 Publication for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

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

Vielen Dank für Ihren Anruf. Ich habe nun eine Antwort aus dem Labor erhalten. Leider haben wir nur Erfahrung mit humanen Zellen für bis zu 4 Stunden in CyGEL. Es wird allerding dringend empfohlen das Gel mit Medium und anderen benötigten Zusätzen zu versetzen um die Zellen gesund zu halten. Hierbei muss beachtet werden, dass die Zusätze hoch konzentriert sind und dem Gel nicht mehr als 20% v/v Volumen zugesetzt wird. Weniger ist besser. Wir können leider auch keine Aussagen über das Zellwachstum von Zellaggregaten (wie bei Ihren Insektenzellen) machen, da das Gel die Zellen relative fest umschließt und deshalb das Zellwachstum theoretisch behindert ist. Für Zellaggregate empfehlen wir, die Zellen absinken zu lassen oder sie ab zu zentrifugieren auf die optische Ebene. Ich hoffe, diese Information was hilfreich. Ich würde mich sehr über ein Feedback von Ihnen über ab109205 freuen. (We don't have data on insect cells in CyGEL, and our experience is limited to human cell lines immobilised for ca. 4 h. It is important to supplement the CyGEL Sustain product (aqueous) with appropriate culture medium (CM) and any other additives at a high concentration (e.g. 10X) to avoid the over-dilution of the gel. Over-dilution would lead to failure to gel. The total dilution should be limited to 20% v/v. The goal is to have the gel be 1X for CM after all additions. A viability dye might also be added as required. We cannot confirm whether the cell clumps will be able to proliferate in CyGEL or not. Typically, cells in CyGEL are held very firmly in place and this would tend to limit cytokinesis. It may be helpful to allow the clumps to sink or to spin them down to the optical plane.)

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Abreviews
The CyGEL sustain is a good gel to investigate suspension cells in an environment similar to the cell normal conditions. For microscopy purposes is a good approach since is optically inert. For the purposed I used, I employed a modified protocol than the provided one. This affects the time the gel last in it best conditions. Under these circumstances I could work with the gel for 3-4 hours without degradation. Noteworthy, one should carefully consider the properties before using it, as well as the environmental conditions where they plan the experiment. The osmolality needs to be considered, and for this purpose there is a special vial that comes along with the kit. But additionally, the temperature and humidity conditions, because the gel under dry conditions disintegrates faster. Therefore in physiologically relevant conditions the gel last less than what is specify in the brochure. The CyGEL sustain performs better under this conditions than the CyGEL.

Abcam user community

Verified customer

Submitted Mar 31 2015

Answer

I can confirm that CyGEL (ab109205) is entirely synthetic and it contains no biologically-derived material.



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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

Thank you for contacting Abcam.

We do not have any data for this product being used in a 96-well format but that does not mean to say it will not work, just that we have not yet tested it in that format.

I am sorry that I am not able to provide more information for you about this, but if there is anything else I can help you with, please let me know.

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Question
Answer

As requested, I have issued a free of charge replacement for one kit of ab109205. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

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