We are requesting funding to purchase Celigo S Imaging Cytometer, an advanced multi-purpose cell imaging and quantitation instrument, to enhance and expedite cancer cell culture studies at the Canary Center for Early Cancer Detection at Stanford. Accurate cell culture studies are an important and necessary component of nearly all cancer early detection and therapy studies. However, we still perform cell culture analysis semi- manually by counting a small aliquot of trypsinized or suspended cells (10-20 ?l) using a hemocytometer and brightfield microscope and extrapolating the count to estimate the total number of cells. The fundamental problem of this process lies on the use of techniques such as extrapolation, which inherently introduces intra- sample variability while trypsinization affecting cell growth and other downstream assessments. This method beyond being time consuming limit the accuracy and throughput, which are important for improving the success of our studies. We also use fluorescence microscopy-based assays to image and count cells for various applications including cancer cell viability, proliferation, apoptosis and death, transfection efficiency, protein expression, stem cell properties, colony formation, cytotoxicity, drug dose/response and treatment efficacy studies. The current methods are not optimal for our research, as these assays require real time visualization under microscope and manual digital analysis for collecting final results. Due to prolonged exposure time of cells under microscope, the cells are exposed to sub-optimal conditions that affect cell growth with occasional photobleaching of the expressed fluorescent reporters. Alternatively, the proposed commercially available Celigo cell imaging system integrates both advanced brightfield and fluorescent imaging for cell counting and quantification. The system is designed to fundamentally improve the accuracy and time of cell culture analysis. The system counts all cells in the entire well without trypsinization avoiding the need of replicate multiple wells to monitor cell growth over time. The optics and scanning technology of the system allow for fast well edge-to-edge uniform cell counting providing efficient and enhanced cell culture studies. The system comes with five channels (one brightfield and four fluorescent) to simultaneously perform measurements over multiple wells. It also comes with user-friendly but comprehensive data acquisition and analysis software including several preconfigured for specific applications. The system with its integrated functionality and advanced software tool, will play a key role for all users at the Canary Center conducting research on developing biomarker based blood tests and molecular imaging approaches to detect and localize early cancers through both in vitro and in vivo diagnostics. As we had a chance to evaluate the instrument through a demo at our center, we are confident that this instrument will provide significant impact to our current research mostly NIH funded by facilitating acquisition of high quality data.
Cell culture experiments are an important early step necessary to collect information about state and conditions cancer cell prior to preclinical and clinical translation of cancer early detections and therapies. Studies that fail in cell culture experiments rarely succeed in in vivo preclinical models and/or in clinical studies, therefore, it is important to obtain accurate and reliable results in cell culture studies using advanced techniques and technologies. The Celigo S Imaging Cytometer requested by this application falls within this category of instruments that helps in collecting high quality data for the NIH funded researches presented in this application which are of highly public health relevant and aligns well with NIH?s mission of advancing human health through fundamental basic scientific discoveries.