Immunofluorescent staining offers a range of benefits over conventional immunostaining, including: higher signal: noise (S:N) superior spatial resolution specific co-labeling of multiple objects in the same sections and direct correlation between fluorescence intensity and antigen concentration in tissue sections.ĭuring Customer Discovery interviews in preparation for deployment of our AI-based Fully Automatic Stereology Technology (FAST®), a significant number of current and future SRC customers evinced an interest in expanding this NSF-funded approach to automatic quantification of fluorescent images. The m=1 Gundersen CE value is the newer method, and is recommended.Background of Fluorescence and Stereology Integration Stereo Investigator calculates the CE estimates with both smoothness class equations m=0 and m=1 because it is not possible to know from the samples which method is appropriate. If you want to compare your results with older papers published using the pre-1999 CE estimate, you may want to use the m=0 smoothness class for a more direct comparison. The efficiency of systematic sampling in stereology and its prediction*. In summary, the m=0 Gundersen CE equation was the original CE estimate developed for use with the optical fractionator (cf. Gundersen, H. The efficiency of systematic sampling in stereology-reconsidered.
#Stereology ce full#
For a full explanation, please refer to: Gundersen, H. The authors determined that biological tissues are best described by the m=1 class. published a new paper referenced below, which reconsiders the CE issue and recommended estimating the CE with the m=1 smoothness class. Estimators of the precision of stereological estimates: an example based on the CA1 pyramidal cell layer of rats. That is why m=1 is recommended and usually used today.įor a case in which the m=0 CE seems to be more appropriate, please refer to: Slomianka, L., & West, M. Sudden cutoffs are uncommon in biological tissues. Sections closer to the top of the heart show increasing area until the end of the chambers is reached. Near the tip of the heart the chambers are small and so is the cross sectional area. The volume of the chambers of a heart can approximate a triangular graph when the heart is sectioned perpendicular to its long axis. A triangular graph is 0 until it ramps up in a straight-line segment and suddenly plummets back to 0. Suppose that the samples come from something that has a triangular graph. The easiest way to understand the difference is to consider an example of sampling in which the m=0 result is appropriate. Notice that the decision to use m=0 or m=1 is made outside of the obtained samples. However, if there are any sharp cutoffs in the data, then the m=0 results are the appropriate results to use. The authors decided that most biological tissue has characteristics that are described by the m=1 results. The m=0 and m=1 results differ in some arcane mathematical descriptions that relate the collected samples to a model of the object from which the samples were obtained. The latter is done by estimating the CE.Īll CE estimates are model based and the CE method published by Gundersen is no exception. The second is to get an idea of how good the estimate is. The first is the estimate of interest, be it cell counts or any other geometric quantity. The stereological sampling process has two considerations.