Mixed Models - remedy for pseudoreplication?
Dear List readers, we are studying effects of chemical therapies on the proliferation of cancer cell lines. My question aims at the practical application of a Mixed Effect Model (lme in R) of a particular analysis of a cell culture experiment on a 96 well plate and the problem of pseudoreplication. A 96 well plate looks like this: http://www.cellsignet.com/media/plates/96.jpg and receives treatments in the columns 3-10 with 8 wells, i.e. the replications. We repeated the experiment on 6 days, with one 96 well plate per day. We noticed a high intra-day variability. Since all the cells of one day on which the treatments are applied come from the same flask and the media and pharmacologic agents, with which they were treated, are coming from the same tubes, the question arises if I have to count them as pseudoreplicates. The analysis of the data using ?lme? works just fine, but I worry about the high number of degrees of freedom, which include the ?pseudoreplicated? measurements. An alternative would be to average the 8 replicates per day and use an ANOVA with an Error structure, which seems less elegant to me, since we are not including the high intra-day variability. I would be very happy if anyone could give an adivce on how to deal with this dataset. I found different opinions in the literature, some saying that LMMs are a "remedy" for 96 well plates and some say that 96 well plates are per-se pseudoreplicated... thanks in advance! Jens
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Dr. Jens Oldeland
Post-Doc Researcher & Lecturer @ BEE
Managing Editor - Biodiversity & Ecology
Biodiversity, Ecology and Evolution of Plants (BEE)
Biocentre Klein Flottbek and Botanical Garden
University of Hamburg
Ohnhorststr. 18
22609 Hamburg,
Germany
Tel: 0049-(0)40-42816-407
Fax: 0049-(0)40-42816-543
Mail: jens.oldeland at uni-hamburg.de
Oldeland at gmx.de
Skype: jens.oldeland
http://www.biologie.uni-hamburg.de/bzf/fbda005/fbda005.htm
http://www.biodiversity-plants.de/biodivers_ecol/biodivers_ecol.php
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