Objective quantitation of serum antibody titres against Enterobacteriaceae using indirect immunofluorescence, read by videocamera and image processing system

Abstract

Objective quantitation of serum antibody titres against Enterobacteriaceae using indirect immunofluorescence, read by videocamera and image processing system Apperloo-Renkema, H. (1991). Objective quantitation of serum antibody titres against Enterobacteriaceae using indirect immunofluorescence, read by videocamera and image processing system. Default journal. CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Summary. A new way of measuring indirect immunofluorescence (IIF) of microscopic bacterial slide preparations by videocamera and an image processing system is presented. This method is compared with the conventional method of reading the slides by eye. The advantages of this new approach are objective reading, greater accuracy and easier performance. We have applied the method to measure serum antibody titres against endogenous Enterobacteriaceae. The method offers the opportunity to combine IIF with automatic morphological analysis, thereby maximally exploiting the possibilities of the immunofluorescence technique.

Circulating antibodies against faecal bacteria assessed by immunomorphometry: combining quantitative immunofluorescence and image analysis

“…When two faecal samples are compared, it is important to know per morphotype which differences in titre are significant. The difference between duplicate measurements was considerable compared to the differences between duplicate measurements in pure cultures [9]. For IgA the mean absolute difference (S.D.)…”

“…The slope and intercept of the resulting linear fit were interpreted as indicative of antibody concentration and background brightness level, respectively. To calibrate the slope, a threshold at which the human eye observes positive fluorescence, was determined as described previously [9].…”

“…We have previously shown that in case of pure cultures of Enterobacteriaceae, it is possible to assess serum antibodies by indirect immunofluorescence. This was achieved by measuring the immunofluorescence per bacterium in a quantitative way by computerized image analysis [9]. By combining the indirect immunofluorescence method and the computerized image processing, one has a tool for the assessment of the (plasma) antibody response of several isotypes against morphologically defined groups of bacteria ('morphotypes'), in an objective way.…”

Quantitative Measurements of Intestinal Ecology by Digital Image Analysed Microscopy

“…An important extension of the system used above was made by Apperloo-Renkema et al (2). In this case, the aim was to quantify the systemic immune response to the intestinal microflora, by the addition of immunofluorescence measurement.…”

“…It allows measurement of many flora parameters without the need for culturing. Several applications have been developed in Groningen, initially just to perform automated counting and morphometric analysis (21)(22)(23), later adding fluorometric analysis as well, including immunofluorescence measurements (2,3,(12)(13)(14)(15) and more recently fluorescence in situ hybridization (FISH) (4). In this paper we will review the progress made in the microbiological image processing programme in Groningen.…”