Paraffin embedding and sectioning

[Amritha Varshini]

Hello TooT team!

We just sent off our organoids for paraffin embedding. The core we're using wants to know the thickness of the sections required. Could you let us know what thickness your slides were?

Thanks,

Amritha

Amritha Varshini Hanasoge Somasundara, PhD

Research Assistant Sr.

Levine Lab

Memorial Sloan Kettering Cancer Center

[Abdullah Khan]

Hi Amritha, 

We use 4um thickness slides for most of our paraffin slides (12-15um for OCT/frozen sections). 

FYI we now embed in histogel which makes for very nice even blocks. If helpful I can put up a couple of images of what that looks like? 

Thanks, 

Abs

[Amritha Varshini]

Yes please, that would be great! 

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[Abdullah Khan]

Hi Amritha, 

Please see attached/below + STLs if you want to 3D print the mould we use. I think my autocorrect changed the spelling of mould repeatedly but I couldn't really be arsed to double check! 

Let me know if you have any questions and good luck ! 

Abs

[CAMILLA TOMBARI]

Hello TooT!

Maybe I missed it, but have you shared the STLs to 3D print your mould? It would be greatly appreciated.

Moreover, I would like to ask how do you usually proceed once the organoids are embedded in histogel blocks, do you dehydrate them and then proceed with the hystoclear baths of the whole blocks?

Thank you in advance!

Camilla

[Abdullah Khan]

Hi Camilla, 

Sure thing - see attached STL and .3mf files. You can scale it up or down as it suits you. 

We use the 3D print as a negative and make the mould out of silicone: 

https://www.amazon.co.uk/Elastic-Silicone-Making-Rubber-Casting/dp/B09M9XKPFW/ref=asc_df_B09M9XKPFW?mcid=58476f004e6e3d548ffbff41a976bfe5&tag=googshopuk-21&linkCode=df0&hvadid=696555674561&hvpos=&hvnetw=g&hvrand=3269318667796212627&hvpone=&hvptwo=&hvqmt=&hvdev=c&hvdvcmdl=&hvlocint=&hvlocphy=1007201&hvtargid=pla-1646182265027&psc=1&gad_source=1

Doesn't have to be this, but something similar would work well. 

Once the organoids are in a histogel block, we usually ship them in PBS to a histo lab. But yes, the process is dehydration + histoclear of the whole blocl for paraffin embedding. 

GOod luck and thanks for TooTing! 

Abs