Hello Pieter:

Good morning.

There are whole bunch of antibodies available from Abcam and Novus Biologicals that label microglia. I have not done any experiment with Zebrafish but I purchased antibodies from these 2 companies and trust their quality. There are some links here for you to look at those antibodies. You can also call the customer service to ask if they have any particular protocol available for zebrafish microglia staining.

The links are:

http://www.abcam.com/neuroscience/an-overview-of-microglia-subtypes-and-markers

http://www.novusbio.com/antibody-news/antibodies/the-cd11b-antibody-%E2%80%93-a-marker-for-microglial-cells

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC499969/

Good luck and hope this helps you. Ranu

Dear Pieter,

as far as I know microglial antibodies used in mammals do not label microglia en ZF. Indeed I think there is a isolectin IB4 that is used to label microglia in rat cultured cells  and I read an article where IB4+ cells were identified as microglial cells in the ZF retina.  Zou S, Tian C, Ge S, Hu B (2013) Neurogenesis of Retinal Ganglion Cells Is Not Essential to Visual Functional Recovery after Optic Nerve Injury in Adult

Zebrafish. PLoS ONE 8(2): e57280. doi:10.1371/journal.pone.0057280

Cheers

Paula

First off, thanks to both of you for your helpfull comments. I am familiar with the companies you mentioned Ranu and I have had good experiences with them. However, as Paula said a lot of antibodies for mammals do not work in zebrafish.

Thank you Paula for the paper and the suggestion. I will look into the possibility of using IB4 antibody in zebrafish. I will let you know if it works out.

Best,

Pieter

It is not an antibody is a lectin coupled to a fluorophore (Isolectin IB4-with alexa or FITC for instance). It binds to extracellular matrix components, many times on vessels but this particular one in ZF (at least in the retina) it seems to bind to microglia but not to blood vessels.

Good luck !

Paula

Thanks for pointing that out. I have never used lectins before actually but. I will definitely give it a go and see if it works in the zebrafish brain, I'll keep you posted.

Happy holidays!

Pieter

Hi Pieter,

      Is there a specific reason you need to do immunostaining for microglia. I recall Francesca Peri (at EMBL, Heidelberg) had a GFP line labeling microglia, that you can request. I saw beautiful time lapse movies she presented way back in 2008 or 2009 so she may have even more lines by now. If you need to do co-staining you can do a-GFP/fluorescent ISH on transgenic lines. If you want I can send you the protocols we use for this. 

Best

Sandeep

Hi Sandeep,

Thanks for your response. I am familair with this P2Y12 GFP line for microglia labelling. Unfortunately the labelling is a bit weak making it hard for me to use. Your suggestion should solve my problem so I would be very gratefull if you could share your protocols with me.

Best,

Pieter

IBA1 from Wako-Chem Cat: 019-19741 is reported specific for zebrafish

Bommana Raghunath Reddy can you please the ref

Regards