3
$\begingroup$

Please help.

I know how to isolate the different cells (astrocytes, other glial cells, neurons and synaptosomes) from brain tissue using a Ficoll-based separation but how do I determine the purity of the fractions that I have isolated if I cannot use immunological techniques, FACS or SEM analysis.

$\endgroup$
2
$\begingroup$

You can do qPCR for certain markers from the extract. GFAP for astrocytes, Tuj1 for neurons, Myelin for oligodendrocytes etc and calculate their percentages to have an estimate of the relative populations. If you are quite used to microscopy and studying these cells then you can identify the different populations of cells with normal light microscopy as well; this is not quantitative and is susceptible to bias.

$\endgroup$
  • $\begingroup$ q-PCR would be the most accurate PCR technique if you wanted a breakdown of the population by percentage. Simple optical techniques are quantitative if you abide by stereology techniques. $\endgroup$ – user560 Sep 20 '14 at 11:55
  • $\begingroup$ I guess that a Western would count as an example of an immunological technique (excluded in the question). $\endgroup$ – Alan Boyd Sep 20 '14 at 15:41
  • $\begingroup$ yeah I meant qPCR only.. @AlanBoyd.. yeah true 'ill correct it $\endgroup$ – WYSIWYG Sep 21 '14 at 5:26
  • $\begingroup$ @leonardo.. they are quantitative if there is an automated algorithm for scanning and classification of cells based on the images. $\endgroup$ – WYSIWYG Sep 21 '14 at 5:50
  • $\begingroup$ @WYSIWYG - I agree, but that means the technique is not only qualitative. Automated imaging and segmentation is the modern successor to random fields + stereology. $\endgroup$ – user560 Sep 21 '14 at 19:35

Your Answer

By clicking “Post Your Answer”, you agree to our terms of service, privacy policy and cookie policy

Not the answer you're looking for? Browse other questions tagged or ask your own question.