by Susan Hendricks, Ph.D.
Stereology is a powerful tool that can reduce your workload and provide accurate, unbiased quantification. Unfortunately, stereology is often not the most intuitive method for the uninitiated. If it isn’t done properly, the results you attain may not be accurate. Here’s where we can help. Not only do we make Stereo Investigator, the most cited software for stereology, but now we’re offering to do your work for you. Send us your slides, and we’ll complete a comprehensive pilot study for you; providing an objective analysis of your tissue thickness and staining penetration, outlining sampling parameters and counting procedures, and providing you with the knowledge and confidence you need to collect the remainder of the data yourself. Our staff includes experienced Ph.D. research scientists who are experts with stereology methodology, our software, and who have demonstrated experience in neuroscience research applications.
Using a technique created at MBF and adapted from Slomianka and West (2005), we will calculate an adequate sampling fraction based on the section thickness, section interval, and the frequency of the objects to be counted within your tissue. We can also empirically determine the appropriate guard zones for your tissue. This data will be provided to you with the return of your slides in a report detailing how the parameters were chosen. Additional support as you begin your work is also available; we can walk you through your first counting procedure or even audit your work to ensure that data collection is proceeding as expected. Confidence in the technique will translate to confidence in the results.
Consider using our pilot study service as a way to optimize and streamline the quantification for all the animals in your study. And by using MBF to acquire the data of your pilot study, you are saved the time and labor involved in oversampling, to free you and your staff to complete other important experiments. Our research staff scientists and stereology technicians will work with you to outline the histology requirements for your experiment. Based on your hypothesized result, the number of animals needed for the pilot study will be determined. Do you expect to see a large, robust difference between control and experimental groups? Provide a subject from each group to ensure that the sampling is as efficient as possible. Need to remain blind to condition? The stereology pilot study should be performed, at a minimum, on the group in which you expect the fewest number of objects. This will guarantee that enough data is collected for the remainder of the subjects in the study.
A properly designed stereology pilot study will reduce time, effort, and costs for the overall experiment by optimizing the sampling parameters to identify how many sections are needed (and therefore generated and stained) to obtain a result with adequate precision.
Of course, if you want to outsource your complete study, we would be glad to help you with that as well.
For more information, visit mbflabs.com.
Citation: Slomianka L and West MJ (2005) Estimators of the precision of stereological estimates: an example based on the CA1 pyramidal cell layer. Neuroscience. 136(3): 757.
Susan Hendricks is a staff scientist at MBF Bioscience.
First published in The Scope, summer 2008.