Overview

The study of genomics and proteomics has advanced our understanding of the field of biotechnology tremendously over the past 15 years. The translation from genome to proteome brought with it technological innovation at the diagnostic and functional level. The newest developing field is relating the proteome to cell function and the role of the cell as the final arbiter in production of cellular products - this is the field of Cytomics and is poised to become the integral component joining the gene, the protein and now the cell to the organ system in the biotechnology revolution. Linking cell structure and function will be an important frontier in cellular engineering.

Cytomics is the study of the role of the cell within the context of genomics and proteomics discoveries. It recognizes that genes select for specific proteins, which are produced within or on the confines of cells. Cell morphology and structure often determine function. Function or lack of function allows the organism to operate in a normal or abnormal manner. Future cell and tissue engineering will depend heavily on the nature of Cytomics.

The Cytomics & Imaging Research Core at Purdue University focuses on providing state-of-the-art cell measurement systems, encouraging the developing new cellular detection technologies, and enabling and providing resources for service.

Drug Discovery

The Drug Discovery Shared Resource provides essential services for Purdue Cancer Center investigators whose research focuses on the design, synthesis and evaluation of novel anticancer agents and diagnostics. It provides a highly integrated service for the evaluation of candidate compounds in vitro and in vivo.
Purdue University Cytometry Laboratories
Purdue Cancer Center - Drug Discovery
Molecular Cytometry Facilities

Analytical Cytology

The purpose of the Analytical Cytology Core Facility is to provide a high quality, comprehensive resource for investigators. The core has several high quality flow cytometry instruments that are available for analytical as well as sorting applications. Confocal microscopy is also offered as a service. The instruments allow for simultaneous excitation and emission and have heated stages and 10, 20, 40X high NA objectives. In addition, the core aims to provide a high quality consulting environment for all users, to obtain the maximum benefit from the technology available.
Purdue University Cytometry Laboratories
Purdue Cancer Center - Analytical Cytology

Cellular Imaging

The Bindley Biosciences Center is the driving force on the Purdue campus in advanced cellular detection technologies. The core itself has three functions: research and development of next generation technology; integration of current technologies to create enabling capabilities for current day researchers, and resource facilitation for multidisciplinary research projects that require access to state-of-the art cell analysis and imaging technologies.
Cytomics at Purdue University - Research

Key Technologies

Cytomics brings together biological imaging, cell analysis tools such as flow and image cytometry, computational resources for 3D imaging and bioengineering resources for cellular manipulations. It links molecular and cellular biology with the technologies of cytometry. Cytomics embodies the essence of informatics; extracting the information from dots, traces, curves and scans into a knowledge-base of diagnostics and discoveries. Cytomics utilizes the specialty of cell sorting to isolate single cells, colonies, particles or beads for further analysis or biochemical characterization. Cytomics utilizes microscopy in its many forms; transmission, fluorescence imaging, digital imaging, confocal and multiphoton imaging (MP) and laser dissection microscopy (LDM).

In addition to studying the impact of single cell systems, the Cytomics core has a developing program in the field of High Content Analysis (HCA) and the closely related area of High Throughput Analysis (HTA). These technologies are the current driving forces in pharmaceutical drug development.

Cell analysis & cell separation:

The Core is capable of supporting a variety of cell analysis research needs. Cell analysis using flow cytometry allows the isolation and subsequent analysis of individual cells using multivariate analysis. Advanced multi-laser analysis instruments notably the Epics Altra high speed cell sorter with 4 lasers, including a Coherent Enterprise II UV laser and a Coherent Krypton multiline laser in addition to the air cooled Argon and HeNe laser are present in the core. The instrument can sort cells into sterile environments. In addition to this, the core has an Epics Elite cell sorter which is used for development projects, particularly related to instrument modifications, enhancements or any alterations necessary to perform experiments that cannot be performed in more routine instruments. For analysis, the core has two FC500 6 color dual laser analyzers both with automated 32 tube carousels. Each instrument has multivariate analysis software. An Epics XL 4 color single laser Cytometer is also available for basic routine flow cytometry analysis. For specialized microbial operations, the core has a Bio-Rad Bryte which has a lamp based excitation source and is particularly sensitive to very small particles. A significant analytical software suite is available in the core for multivariate analysis.

Laser Capture Cell Microdissection:

Cell types undergoing molecular changes, such as those thought to be most definitive of the disease progression, may constitute less than 5% of the volume of the tissue biopsy sample. Therefore, microdissection is essential to apply molecular analysis methods to study evolving disease lesions in actual tissue where morphologic characterization is required. The core has an AutoPix Automated Laser Capture Microdissection (LCM) System from Arcturus, combining automated upright microscope architecture, three-dimensional optical control of the dissecting laser beam and the dissected area, non-contact tissue sampling and post-dissection handling.

Biological Imaging:

There are multiple imaging technologies available in the cytomics & Imaging core. Basic imaging using standards high quality microscopes allows high resolution imaging in brightfield or fluorescence excitation. To achieve this the core has a Nikon 1000R upright fully automated fluorescence microscope with a SPOT high sensitivity, high resolution digital camera is also present. A Nikon Labophot upright fluorescence microscope and a Nikon inverted 300 fluorescence microscope with a DVC 250 line-scanner confocal microscope is also available. More advanced capabilities exist in other confocal microscopes which can create 3D images using optical slicing. Systems are available to do UV through near IR imaging. A Bio-Rad MP 1024 UV/vis confocal microscope is mounted on Nikon inverted microscope. This unit is ideal for UV live cell imaging. Also available is a Bio-Rad Cellmap 2 color confocal microscope mounted on a Nikon E800 upright microscope. The unit has a 405 Violet laser and a 488 blue diode laser for excitation. The most advanced capability in fluorescence imaging is a Bio-Rad MP 2100 Multiphoton microscope that has the capability of advanced live cell imaging, including live embryos, deep tissue imaging and even functional brain imaging. This instrument also have a Becker & Hinkl life-time module linked to two high speed PMTs that allow for simultaneous Multiphoton imaging and lifetime imaging.

Analytical Software:

The core has a large range of software packages for advanced and basic image processing. These include advanced packages such as Metamorph and Image ProPlus. In addition, an extensive capability exists with the package VoxelView.

High resolution Imaging:

Atomic Force Microscopy specifically for biological samples provides the highest possible resolution for live cell systems. Instruments specifically designed to study biological systems operate under very different conditions to similar technologies operating the area of materials sciences. AFM instruments are available in the core to provide high resolution analysis specifically for biological samples.

Multispectral Imaging:

The latest technology under development in the center involves multispectral imaging. We currently have several instruments available for use that are testbeds for new technology development. For example we have an AOTF (Acousto Optical Tunable Filter) on an upright brightfield/fluorescence microscope. This unit can collect up to 400 images at 1.5 nm increments from 370 to nearly 800 nm. This allows the analysis of multispectral data sets for the potential isolation of autofluorescence or specific target oriented molecules of interest.

Advanced Image Processing:

The core provides multidimensional image processing - 1D, 2D, 3D and 4D image analysis using sophisticated software programs. These packages are directly linked to the detections systems and can extract complex structures and reproduce them in multidimensional space. Such analytical tools are both qualitative and quantitative. We have several packages that are high level image analysis packages such as Image ProPlus, Metamorph and VoxelView.

High Content-High Throughput Systems:

The core is developing capabilities in high content and high throughput analysis typical of systems where thousands of samples must be analyzed daily. These systems require advanced computational support to provide analysis. Current technologies include microtiter based fluorescence imaging instruments which use sensitive cameras and advanced image based computational analysis.

Protein Profiling:

The Core has a unique resource in a state-of-the-art automated, two-dimensional fractionation system expressly designed for high-resolution analysis of complex protein mixtures. The ProteomeLab PF 2D provides an integrated solution that effectively resolves the thousands of proteins present in cell lysates. The ProteomeLab PF 2D system generates data from two dimensions, detailed protein maps can be constructed for easy comparison using the ProteomeLab Software Suite supplied with the system. Liquid fractions can be collected and stored for future analysis or the eluent can be connected directly to ESI-mass spectrometry. The PF 2D's unique combination of chromatofocusing followed by non-porous reverse phase chromatography provides ultra-high resolution of proteins, while delivering information that can be related to 2D gel data. The ProteomeLab PF 2D's second dimension is capable of resolving hundreds of proteins in great detail from each first-dimension fraction. Fraction collection from the first dimension is quality-controlled with an in-line pH monitor, then automatically injected into the second dimension. As a result, PF 2D requires much less time and attention than traditional labor-intensive techniques. The entire 2 dimensional process can be performed in less than 20 hours. Liquid fractions can be stored or transferred to a MALDI plate spotter, or directly connected to an electrospray source.

Consulting and Data Processing:

The Core provides excellent expertise for the design, preparation and analysis of experimental profiles. Consultants are available to advise researchers in the correct techniques; training opportunities are available on a regular basis.