04/10/2009 12:24

Have You Seen Our Article in MedLab Magazine?

CTL scientists have had one of their articles published in a recent issue of MedLab magazine. In this article, they discuss new technology that enables high-throughput microbial detection. They al... [more...]

04/03/2009 11:36

CTL Writes Book Chapter on Validating Reference Samples for Use in Cell-Based Assays

In a new book on cell-based assays, CTL discusses key considerations in validating an ELISPOT assay. [more...]

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CTL QC Sets™

CTL’s Reference Sample QC Sets™ offer outstanding inter-assay control. Use CTL's QC Sets™ not only for validation and protocol setup purposes, but also as internal controls for clinical trials...

CTL QC Sets™


Plaque and Colony Counting


Automated colony counting with the BioSpot line of instrumentation and software.

The CTL BioSpot™ platform contains a wide range of automated image capture and analysis systems for microbial and mammalian colony formation assays. The BioSpot™ line of instrumentation and software is highly adaptable and meets the needs of a broad range of colony counting applications.  These include microbial load and bioburden testing, clonogenic assays, stem cell assays, Ames test, mouse lymphoma assay, viral plaque assays, and more.

BioSpot Main Features:

  • Compatible with all plate formats from 100 mm Petri dish to 96 well microplates.
  • Proprietary illumination enables detection of colonies as small as 25 micro meter diameter.
  • Ideally suited for rapid high-throughput operations.
  • Compliant with 21 CFR part 11 and GLP regulations when operated with BioCompliance™.

BioSpot Colony Counters vs. Manual Microbial Colony Counting

Compendial methods for microbial load and bioburden testing specify the determination of colony forming units (CFU) per gram of sample. Traditionally, plate counts have been determined by manually counting colonies within 60-100 mm Petri dishes. This is naturally a tedious, error prone and time consuming process.

BioSpot colony counters enable automation and/or miniaturization, transforming CFU determination into a quick, easy and accurate process. This provides an easily validated adaptation of compendial methods.

BioSpot™ Colony Counting vs. FACS

The BioSpot™ platform enumerates groups of cells growing as colonies and can readily distinguish colonies against a background of single cells.  By definition, colonies are clusters of cells grown on or in a solid support.  In contrast, fluoresence-activated cell sorting (FACS) can only enumerate single cells in suspension.  This precludes the use of FACS/flow cytometry for counting colony formation assays.  The ability to identify true colonies is critical for the analysis of both microbial and mammalian colony formation assays. 

BioSpot Applications

Microbial assays

The ability to rapidly and reliably enumerate bacterial colonies is paramount for microbial load and bioburden testing in product safety markets.  These include the food and dairy industries, environmental screening (water and air), personal care product testing, toxicology screening, and pharmaceutical testing.

Conventionally, these tests have been conducted manually with minimal throughput and high error rates.  Recently though, there has been an extensive effort to develop alternative (rapid) methods to replace the slow and tedious traditional methods.  The BioSpot™ product line enables miniaturization and automation of the standard plate count, thereby producing a dramatic increase in throughput and reproducibility.

Mammalian colony assays

Mammalian colony formation assays are an important tool in oncology and stem cell research, both of which rely on manual enumeration of colonies.  Clonogenic assays, for example, are typically used for testing ionizing radiation and drug treatments for cancer therapeutic potential.  Stem cell assays are used to identify multi-potent progenitor cells from bone marrow, cord blood and peripheral blood.

Mammalian colony formation assays rely on the ability to distinguish colonies from a background of single cells.  The BioSpot™ product line enables colonies to be size-selected for enumeration, thus only counting true colonies.

Viral plaque assays

Localized lysis of virus-infected cells in a confluent monolayer produces a plaque that can be detected through direct staining or by staining cells in the monolayer.  Viral plaque assays utilize either a monolayer of mammalian cells or a bacterial lawn.  The size and morphology of viral plaques is highly variable and notoriously difficult to detect and count.

The BioSpot™ product line conducts multiple object-oriented morphometric measurements, enabling user-defined gating to separate plaques from a complex background.  This compensates for the inherent variability in plaque assays and enables reliable enumeration.

Genotoxic assays

Genotoxic assays are used to measure mutagenic potential by detecting forward or reverse mutations with selective media.  Genotoxic assays use either mammalian cells (ie: mouse lymphoma assay) or microbes (ie: Ames test) and are easily imaged and counted using the BioSpot™ system.  

 

What to learn more? Click here to download a copy of our BioSpot™ applications flyer.