Sensitivity gain with 20 um ID nano-columns over 75 um ID

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Sergo
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Sensitivity gain with 20 um ID nano-columns over 75 um ID

Postby Sergo » Wed Apr 24, 2013 10:51 am

Hello all!
Anyone here tried doing nano-LC with 20 um ID and 20-50 nl/min? Have you seen a significant sensitivity gain over the conventional 75 um ID and 200 nl/min and...would this be worth the trouble..clogging on every possible occasion.
Cheers,

Sergo

cavaliersun2003
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Postby cavaliersun2003 » Tue Jun 11, 2013 4:37 am

It will be interesting to see if 20 um ID works out. Ideally, smaller internal ID means lower dilution effect, and the sensitivity is directly proportional to the (1/d)^2, in which d represent the internal diameter, but you can load very limited sample to 20 um ID, and I guess your column will clog more than 75 umID.
The other thing I believe, correct me if I'm wrong, is that if only use 20 um ID nanocolumn while other connecting tubes are in bigger ID, then the dilution still happens. In other word, you need to configure the whole LC system, not just the column, to a small ID format.

Christopher
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Postby Christopher » Thu Apr 24, 2014 11:09 am

There was an interesting paper quite a few years ago now that used a 15um ID with 85 cm length columns. With an AMT approach they get enough of a boost in sensitivity to detect proteins from a single cell (http://www.biotechniques.com/multimedia/archive/00037/BTN_A_04374RV01_O_37072a.pdf).

In my experience you do see a nice benefit by moving down to a smaller ID. It is a bit more challenging to pack the columns to suitable lengths, but I do not notice such an increase in blocking provided you scale down your loading amount. In the end I settled with a 40um ID, as it provided a good balance of sensitivity increase while still providing relatively easy packing and good loading amounts.

Would be interested to hear your experience if you try it out.


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