Live Cell Assays, LLC (LCA) is an early-stage, privately-funded biotechnology company. LCA was founded by Daniel E. Callahan, Ph.D., who is the co-inventor of a computer vision method for the automated interrogation and manipulation of living cells. This method is called "Visual-Servoing Optical Microscopy™" (VSOM™). VSOM™ is a optical microscope digital imaging method. In December 2008, LCA (founded June, 2008) obtained the exclusive commercial license to all VSOM™ method patents. The first US VSOM™ method patent was recently awarded (Assignee, The Regents of the University of California, US 7546210, June 9, 2009). Multiple VSOM™ patents have been awarded in Australia (AU 2007201501, AU 2001266748). Many other VSOM™ patents are pending in countries around the world. The patented VSOM™ method has many potential applications that do not involve living cells. However, LCA is currently focusing solely on the development of VSOM™ Live Cell Assays. LCA is developing VSOM™-enabled instruments, VSOM™ software, VSOM™ fluorescent probes, VSOM™ sterile specimen vessels and other innovative devices for quantitative optical microscope digital imaging. Our products will facilitate the automated discovery and optimization of a new class of live cell assays we call VSOM™ Live Cell Assays.

A New Class of Live Cell Assays

VSOM™ is a method for the automated discovery, application and interpretation of a new class of live cell assays. VSOM™-enabled instruments and VSOM™ Live Cell Assays facilitate the automated and long-term monitoring, testing, and physiological manipulation of large numbers of individual living cells. One example of a VSOM™-enabled instrument is our third generation prototype, VSOM™ Beta (see images and animation snapshots on Home page). VSOM™ Beta is built around a modern, automated optical microscope. VSOM™ Beta establishes a direct fluidic coupling between computers and large numbers of individual living cells. This fluidic coupling is made possible by the use of multiple computer-controlled syringe pumps (see our Technology page).

VSOM™-enabled instruments and software generate, interpret and modulate live cell physiological responses. The VSOM™ Beta instrument monitors multiple fields (and multiple focal planes) of individual living cells in multiple digital image channels at approximately 5 minute intervals. Multichannel fluorescence and transmitted light (including phase contrast and DIC) digital images are acquired in any combination at each image acquisition time point. Changes in cell shape and location are computed and recorded at each time point. Calibrated fluorescence intensities in specific subcellular regions of interest are also computed and recorded at each time point.

Physiological Response Phenotypes

Subtle differences in physiological response phenotypes are the basis for VSOM™ Live Cell Assays. VSOM™ is an emerging technology that will enable the automated discovery and implementation of a wide range of gentle live cell assays useful for the detection and long-term monitoring of subtle changes in the physiological state of individual live cells. VSOM™ Live Cell Assays allow VSOM™ system software to sense and interpret changes in the physiological status of living cells. VSOM™-enabled instruments, system software, and specimen vessels allow researchers to develop custom collections of VSOM™ Live Cell Assays in an automated manner.

Automated Experiment Control Decisions

VSOM™ system software monitors physiological responses on a cell-by-cell basis. Detection of a small difference in the physiological response of a single (perhaps rare) cell can trigger an automated (i.e., software controlled) experiment control decision. One example of an automated experiment control decision is a software decision to modulate the on-going physiological response of one or more cells. Modulation of physiological responses is one way to discover or optimize new VSOM™ Live Cell Assays.

Another example of an automated experiment control decision is a decision to immediately and automatically select and apply a second VSOM™ live cell assay to the same population of cells. Selection (from a large collection of VSOM™ Live Cell Assays) and application of the second assay is based on automated interpretation of results produced by the first assay.

A third example of an automated experiment control decision is a decision to alter the microenvironment of the specimen vessel until certain cells (possibly a small subpopulation of cells) achieve the desired physiological state or biological endpoint. In this case, on-going read-outs of the current physiological states of individual cells are provided by repeated application of the appropriate VSOM™ Live Cell Assay.

Modulation of Physiological Responses

Computer-controlled modulation of single cell physiological responses gives VSOM™-enabled instruments and VSOM™ system software an ability to discover and optimize subtle differences between physiological response phenotypes. The ability to detect subtle differences in single cell physiological responses means that the computer-applied stimuli and other perturbations necessary to generate a physiological response can be exceptionally gentle. Optimized and gentle VSOM™ Live Cell Assays can be applied repeatedly and in various combinations. Our products facilitate the automated discovery and optimization of optimized and gentle VSOM™ Live Cell Assays. Our products will allow research scientists to rapidly develop and share large collections of VSOM™ Live Cell Assays that are uniquely suited for a particular research application.

VSOM™ Live Cell Assays can be developed for any single cell physiological response that is detectable via single or multichannel digital image analysis. For instance, VSOM™ Live Cell Assays that employ multiple color fluorescent probes (including fluorescent proteins) can be developed. In addition, multichannel assays that combine transmitted light and fluorescent light assays on separate digital image channels can also be developed.

Animation 1: Automated Assay Development

Animation 1 (see Technology page) illustrates some important aspects of automated VSOM™ Live Cell Assay development on theVSOM™ Beta instrument. In this simple overview of the VSOM™ method, changes in single cell morphology are monitored and quantified as VSOM™ system software makes automated experiment control decisions. The system software applies various microenvironmental perturbations by selecting and activating one or more computer-controlled syringes. The experimental objective is the discovery and/or optimization of unique physiological response phenotypes (time-dependent changes in morphology) that can discriminate between different cell types. In this particular animation, "physiological response curves" represent (generic) changes in cell morphology as a function of time. However, VSOM™ Live Cell Assays can be developed for any live cell physiological response that is detectable via single or multichannel digital image analysis. Animations 2 and 3 will illustrate the technical details of automated VSOM™ Live Cell Assay development.

Animation 2: (coming soon)

Animation 3: (coming soon)