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Biological Science Applications
Colony Counting
The Sorcerer Automatic Colony Counting System utilizes an innovative Petri-viewer or Macro-viewer to ima
ge various types of culture dishes, flasks and multi-well plates to provide fast, accurate counts in real time.
The system has powerful image analysis software including facilities for separating overlapping and clumped colonies as often encountered in mammalian cell lines. A high resolution version of the system is also available for particularly demanding situations. The system is in use in pharmaceutical and biotechnology companies, universities and hospital laboratories.
Applications include:
- Ames Test (Bacterial Reverse Mutation Test)
Named after its developer Dr Bruce Ames, it is the most widely used test for genotoxicity, i.e. the ability to induce mutation or cancer. Testing for genotoxicity is mandatory for pharmaceuticals, medical devices, food additives, agrochemicals and industrial chemicals Sorcerer provides rapid counts of revertant colonies. The choice of incident, transmitted & dark-field illumination facilitates counting on all combinations of test substances in Vogel-Bonner or alternative media.
- Clonogenic assays using stained or unstained colonies in 6, 12, 24, 48 or 96 well plates are rapidly read by Sorcerer with a Macro-viewer and Automatic XY stepping stage. The stage can be programmed to scan all or part of each plate.
Results are presented in an Excel spreadsheet as colony count per well or as a complete colony size distribution per well.
- Mammalian Cell Colony Counting, Coliform, TVC, Spiral plates, Plaque counts, 3M Petrifilm and traditional Pour and Spread plate counts.
Chemotaxis
The quantification of eukaryotic cellular migration has been greatly aided by image analysis technologies such as the Sorcerer image analysis system. One of the most popular methods for studying cell migration in response to stimulatory agents involves the movement of cells through pore-containing membrane filters.
In membrane filter assays, two commonly used filter types are polycarbonate and cellulose nitrate. Polycarbonate membranes, which are only 10 µm in thickness, are substantially thinner than cellulose nitrate membranes, which are 100-150 µm in thickness. Cells that migrate through polycarbonate filters have a relatively short distance to move in order to pass through the filter, and all the responding cells are found in one optical plane of the filter. In contrast, cells that migrate into cellulose nitrate membranes do not move uniformly through the filter to the opposing surface, and consequently cells are found in numerous optical planes throughout the filter.
The Sorcerer Image Analysis System utilizing a high sensitivity CCD camera coupled to a light microscope produces a live video image of stained cells on the membrane filter. The program is able to detect stained cells in the video image by virtue of contrast differences between the cells and the filter background. The system automatically counts cells that have migrated through thin polycarbonate membranes, or partially through thick cellulose nitrate membranes.
Cell count data is transferred in real time to a Microsoft Excel spreadsheet where data tables and graphs showing cell migration in response to stimulatory agents can be generated
ELISPOT (enzyme-linked immunospot assay)
The enzyme-linked immunospot assay, or ELISPOT assay for short, is used to determine cytokine se
cretion by individual B-cells or
T-cells following in vitro stimulation with a specific antigen, which provides a measure of cytotoxic activity in viral, bacterial and tumor studies. The assay is often used in the research of the HIV and AIDS viruses.
The Sorcerer image analysis system used in conjunction with a Macro-viewer and Automatic XY stepping stage to automatically position the 96-well microtiter plate on each well to be measured.
Results can be presented in an Excel spreadsheet as spot count per well or as a complete spot size distribution per well.
Advanced correction techniques and a powerful matrix detection algorithm help separate touching spots and compensate for uneven illumination within wells. A high resolution version of the system is also available for particularly demanding situations. T
Inhibition Zones
Anti microbial bioassays to determine the efficacy of antibiotics are conducted by public he
alth laboratories and in the pharmaceutical, veterinary and medical device industries. One of the most widely used methods is to culture bacteria in the presence of the antibiotic, usually applied as an impregnated disk, or in a well cut into the agar. The resultant zone of inhibition, i.e. the region around the disk or well where growth has been prevented, is then measured. Traditionally, this has been done by manual or electronic calipers, but today is achieved using image analysis.
The Sorcerer image analysis system combined with a purpose-built Macro-viewer is ideal for the measurement of inhibition zone bioassays . The viewer can accommodate 100 mm & 150 mm Petri-dishes as well as large format 12" square culture trays. It has various illumination techniques enabling it to be used for opaque or transparent culture media.
Inhibition zones are measured by virtue of the difference in contrast between the bacterial lawn and that surrounding the disk or well where growth has been prevented. Special algorithms for hole-filling and edge-enhancement ensure accurate measurement of zone diameter or area. Zones can be analyzed singly, at high magnification, or as multiple zones using programmable scanning measurement frames. Data can be sent directly to Microsoft Excel, or to Microsoft Access or Oracle databases for further processing.
Mouse Lymphoma Assay
The L5178Y mouse lymphoma assay, using the thymidine kinase (tk) gene, is the most widely used in vitro mammalian cell gene mutation assay. There are two methods for performing the assay, one using soft agar medium for cloning and enumeration of mutants, and the other using liquid medium and 96-well multiwelll plates.
The Sorcerer Automatic Colony Counting System is an ideal solution for the Soft Agar version of the Mouse Lymphoma Assay. The system uses a purpose built Petri-viewer to obtain a high contrast image of the culture dish. Sorcerer produces a complete size distribution based upon colony Area or Diameter. The system has the ability to save data to Microsoft Excel, Microsoft Access and Oracle databases.
Multipoint MIC & Antibiotic Sensitivity
Perceptive Instruments has worked for many years with public health agencies and contract research
organizations to provide data processing and automatic plate reading solutions for multipoint microbiology. In particular, we have developed custom solutions for antibiotic susceptibility testing and determination of minimum inhibitory concentration (MIC).
The Sorcerer image analysis system, linked to a purpose built Macro-viewer is the ideal solution for multipoint microbiology. The macro-viewer has a range of illumination techniques enabling plates with clear, opaque or semi-opaque culture media to be analyzed. The system automatically measures colony growth at each pin position, with the whole plate being analyzed in seconds. Multiple pin formats on petri-dishes or rectangular culture plates can be accommodated.
Unscheduled DNA Synthesis (UDS)
The Sorcerer image analysis system linked to a light microscope is ideal for scoring slides a
nd processing the nuclear and cytoplasmic grain count data using Microsoft Excel macros. Off the shelf and customized solutions for UDS based around the Sorcerer system can be provided
Customized packages consist of a series of modules for creating studies, randomization, slide scoring, data saving and production of experimental summaries. Slide scoring can be based either on direct or indirect counts and the analysis of one or more cytoplasmic areas. An operator scoring the slides is guided throughout the procedure, including calibration where necessary, and all events are fully audited.