Frequently asked questions (FAQ) about unbiased stereology for bioscientists. Click on the question to review the answer.
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Student discounts for neurostereology workshops?
Are student discounts available for the Fall Neurostereology Workshop?
We support student training in several ways. Our merit-based Stereology Fellowship Program offers full waiver of registration fees. Also, early registration at workshops includes reduced registration fees and sleeping rooms at discounted rates.
Effect of cells in clustered distribution on count?
The cells in my reference space have a somewhat clustered distribution. Does this affect accuracy of my Stereologer counts?
A more or less clustered distribution does not affect accuracy, but may require that you increase your sampling stringency. In a pilot study, the Stereologer will identify the sampling level (animals, sections, cells) showing the greatest variability and suggest changes to your sampling parameters, e.g., decrease director spacing to 300 um, for the rest of your cases. In this way, the system automatically optimizes your sampling scheme for maximum efficiency.
What is meant by the "Corpuscle Problem?"
What is the Corpuscle problem?
The Corpuscle Problem is the bias (systematic error) introduced by attempting to quantify the number or density of 3-D objects (cells) based on the number of 2-D profiles on a standard histological section. This bias occurs because cells with a larger size, more complex shape, and/or their long axis perpendicular to the sectioning direction are more likely to appear in the cell count, which leads to a biased count. The disector principle introduced by Sterio (1984) avoids this problem by counting all 3-D objects regardless of their size, shape and orientation.
My cells of interest have a somewhat clustered distribution. Does this affect accuracy of my counts?
A more or less clustered distribution does not affect accuracy but does require adjusting your sampling strategy to achieve maximal efficiency. In a pilot study Stereologer automatically identifies the major sources of variability in your data and adjusts your sampling parameters accordingly. In this way the system optimizes your sampling effort for maximum efficiency, which dramatically increases the throughput of your studies.
Caveat emptor (latin, "let the buyer beware")
We often receive queries from people new to the field about why Stereologer is the best choice for computerized stereology. Our answer is because the Stereologer is:
- the most comprehensive and competitively priced computerized system available.
- designed by a team that has published more than 300 peer reviewed stereology papers and helped customers publish many times that number in a wide variety of disciplines.
- supported by Stereology Academy, which has taught the principles and practices of unbiased stereology to more than three thousand bioscientists since 1995.
- the only commercially available stereology system that runs native (i.e., without virtualization) on both PC and Mac operating systems.
We encourage customers to compare our costs, features, and support with other systems available. To save you time and effort, here are the other computerized systems available and their company names and headquarter locations:
- Mercator: Explora Nova (La Rochelle, France)
- newCAST: VisioPharm (Copenhagen, Denmark)
- StereoInvestigator: MBF Bioscience (Williston, VT)
- BioQuant (Nashville, TN)
When comparing commercially available stereology systems, it helps to keep one thought in mind: Caveat emptor (latin, “let the buyer beware“), which arises from the seller knowing more about the purchased goods than the buyer.
A few specific points to consider:
1) Why are some systems so expensive? To increase profit margins some companies aggressively up-sell and overcharge for hardware and extraneous software. Beware of “gold-plated” computerized systems priced in some cases 15-30% or more higher than the Stereologer.
2) Why isn’t all stereology software user-friendly? The recipe for effective computerized systems calls for about 1 part computer to 4 parts stereology. A programmer with reasonable skills can build a stereology program from published formulas. Without bioscience research training and stereology experience, computer scientists and businesspeople develop “clunky” non-intuitive software.
3) Why can’t all support teams answer my stereology questions? With primary expertise in marketing rather than stereology or even bioscience, aggressive sales reps can master the terminology; however, this thin veneer is easily exposed. Companies without credentials to teach stereology lack the expertise to support stereology-related questions from customers, including issues related to histology, efficient research designs, data interpretation, optimization for maximal efficiency, and biological variability. Instead, all they can do is answer questions about their software.
4) Why do some PC-based program not run well on my Mac using Parallels? Some companies promote Windows-based programs that can run on Mac iOS through a commercially available simulation program (Parallels). Regardless of their sales pitch, running a Windows-based program via “virtualization” can be highly problematic. Check the on-line users groups devoted to sharing solutions and resolving problems with running PC-based programs on Mac via Parallels.
Space balls for fiber length on coronal sections?
My sections are cut in a coronal plane. Can I still use the space balls method to quantify fiber length?
We developed the Space Balls method more than a decade ago (Mouton et al., Journal of Micrscopy, 2002) to address this exact issue. The Space Balls probe is a sphere that includes all integral angles. Therefore, total length and length density may be quantified on sections cut at any orientation without introducing bias.
Support included with purchase of the Stereologer system?
What are the costs for support with the Stereologer system?
All Stereologer systems include initial set-up, orientation, and study set-up by either on-site or remote access. In addition, all systems include three years of no-cost maintenance and support by phone, email, or remote desktop access. This support includes troubleshooting, bug fixes, upgrades, technical support and assistance with study designs.
Are there additional costs?
The cost of a Stereologer system includes three years of no-cost maintenance and support. Once you set it up in your lab, the SRC helps you get started and then the system runs well. the software is extremely user-friendly (not all computerized stereology systems are), which is an important for someone who has never used computerized systems before. If you have a problem, the SRC calls you back right away, fixes your problem, and you’re on your way. No kidding.
What is the Corpuscle problem?
The Corpuscle problem refers to bias due to object size, shape, and orientation. All of these factors can introduce systematic error (bias) when attempting to count cells on stained tissue sections. The optical director first proposed by Gundersen (1986) uses the disector principle (Sterio, 1984) to avoids this bias through the use of thin focal plane optical sectioning.
Can you send us a quote for processing tissue (embedding, sectioning, staining) as well as collection of stereology data?
Our specialists will work with you to develop the best protocol for processing the tissue to visualize your objects of biological interest. We will send you instructions on the optimal methods for removing, fixing, and shipping your tissue. An appropriate number of stained tissue sections will be analyzed blind to treatment group (when possible) using computerized stereology. On completion of the study we will return your tissue and stained slides together with a Final Report containing methods, statistics, results, and references.
My sections are cut in a coronal plane. Can I still use Space Balls to quantify fiber length?
The virtual sphere approach (Space Balls) introduced by Mouton et al. (2002) uses a sphere probe to quantify length of fibers, capillaries, etc. Because a sphere contains all integral angles, tissue sections can be cut at any convenient direction, e.g., coronal, horizontal, selected by the investigator. Prior to the development of virtual spheres, to quantify length tissue had to be rotated around one or more axes prior to sectioning, which seriously upset anatomical landmarking within tissue sections.
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