13 February 2013 15 2K Report

Hello!

May I ask for your recommendations for tissue processing methods for laser microdissection and subsequent RNA isolation?

I can think of at least the following protocols, each with significant drawbacks (and questions):

1) Traditional FFPE sections:

+ easy handling

+ RNA is safe (but see below)

+ good morphology

- RNA is fixed too, so yields are low and only small fragments retrieved

Here, I'm pretty happy with Qiagen RNEasy FFPE kit - any other suggestions, maybe cheaper?

2) Traditional cryosections:

+ fairly good morphology

+ good yields, good quality RNA if everything goes well

- RNA is easily destroyed

- difficult to handle small samples without melting & destroying RNA

I haven't been very succesful with this option.

3) RNALater -> cryosections:

+ RNA is safe

+ good RNA yields, good quality RNA

- poor morphology

- difficult to section

We have problems making the tissues actually freeze for good sectioning - any tricks or tips here?

4) RNALater -> paraffin sections?

I haven't tried this yet, but should be doable through ethanol etc. Found some references claiming that the RNA quality is poor.

5) New commercial innovations like Qiagen/Prenalytix Paxgene Tissue kit, claiming to achieve both RNA stabilization and good morphology.

I haven't tried any of these yet. Pricing is the obvious drawback.

With best regards,

Mikael Niku, PhD

Department of Veterinary Biosciences

University of Helsinki

More Mikael Niku's questions See All
Similar questions and discussions