Automated Spots Reader
Automated Spots Reader - Related Videos
07:51
Introduction to the Microplate Reader
0 Views •
2023
The microplate reader is a multimodal instrument that allows for a variety of experiments to be performed and measured simultaneously. Microplate readers can make absorbance, fluorescence and luminescence measurements. Multiwell plates are integral to the microplate reader and allow for many experiments to be performed at once. Regardless of the assay type, experiments on the plate reader utilize a standard curve to determine the experimental values. This curve uses samples of known...
03:01
Precise and High-Throughput Analysis of Bacterial Growth Using a Microplate Reader
0 Views •
2025
Source: Kurokawa, M., et al. Precise, High-throughput Analysis of Bacterial Growth. J. Vis. Exp. (2017)This video demonstrates the high-throughput quantification of bacterial growth using a microplate reader. It outlines the steps involved in culture preparation, optical density measurement, and analysis of growth dynamics.
06:53
SpOT the Correct Tissue Every Time in Multi-tissue Blocks
0 Views •
Cited by 5 •
2015
The purpose of the Specimen Orientation Tag (SpOT) is to function as an orientation tool to aid in individual tissue identification in multi-tissue paraffin blocks. These protocols demonstrate how it is constructed easily from common, low-cost histology materials and serves as a reliable visual marker in paraffin blocks and sections.
11:28
Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
0 Views •
Cited by 175 •
2015
The preparing and processing of dried blood spots (DBS) for their final analysis are still poorly standardized for most diagnostic applications. To overcome this shortcoming, a comprehensive step-by-step protocol is suggested and subsequently evaluated with regard to its effectiveness for detecting markers of viral infections.
10:45
Finding Your Blind Spot and Perceptual Filling-in
0 Views •
2023
Source: Laboratory of Jonathan Flombaum—Johns Hopkins University In the back of everyone's eye is a small piece of neural tissue called the retina. The retina has photosensitive cells that respond to stimulation by light. The responses of these cells are sent into the brain through the optic nerve, a bundle of neural fibers. In each retina there is a place somewhere in the periphery where the outputs from retinal cells collect and the bundled optic nerve exits to the brain. At that location,...
FAQs
Counting depends on the criteria assigned to each spot. Size thresholds help distinguish genuine localized signals from very small artifacts, while intensity and shape settings determine whether weak or irregular features are included. Adjusting these parameters can change the number of spot-forming cells reported, so consistent criteria are important when comparing wells or experiments.
The system can evaluate spots produced through different detection formats because ELISpot assays may generate either enzyme-linked colorimetric signals or fluorescent signals. In both cases, the relevant response appears as localized spots within assay wells. The reader scans those wells and applies defined image criteria, allowing the signal format to support quantitative cellular or molecular analysis.
In an ELISpot assay, a secreting cell releases a target analyte, such as a cytokine. The analyte is captured near that cell, and subsequent detection produces a localized spot. This spatial relationship allows spot-forming cells to serve as a measure of secretion events, linking the image pattern to cellular immune activity rather than only to total analyte in a sample.
After the assay produces detectable spots, the reader scans the wells of the plate and analyzes the resulting images. Researchers define or apply thresholds for spot size, intensity, and shape, then use those criteria to count qualifying spots. The resulting spot-forming-cell measurements provide an objective record that can be compared across wells or experimental conditions.
It is useful when investigators need to measure antigen-specific T-cell responses, antibody-secreting cells, or immunity induced by vaccines or pathogens. These applications use spot counts to assess localized cellular or molecular responses in assay plates. Automated analysis is especially relevant when many wells must be evaluated consistently, making immune-response measurements more efficient and less dependent on manual counting.
Automated analysis improves throughput by processing assay-plate images efficiently and applies the same defined criteria across wells. This reduces variability associated with manual counting and supports more objective measurement of spot-forming cells. In studies of vaccine- or pathogen-induced immunity, those features help researchers handle larger response datasets while maintaining a consistent approach to quantification.