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Cytometry Laboratories Areas of Interest

Engineering Interactions

Considering that the specialty instrumentation in this facility consists basically of optical components, computers, lasers, and hydrodynamically controlled devices, it is not surprising that biologists and engineers must interact. There are currently several projects in which direct interactions exist at a high level between biological scientists in the facility and engineering scientists.

Recently the Graduate School at Purdue awarded a special initiative grant for a graduate student to enter the program known as the Optical Engineering Instrumentation and Cell Biology Program. This initiative will be expanded to allow students to interact on any engineering project.

Cytometry Areas of Interest

Advanced Concept Spectroscopic Cytometer Project

This project is designed to utilize state-of-the-art spectroscopic ideas in flow cytometry. The use of time-resolved flow cytometry has some advantages over traditional techniques currently used. Primary interactions are with the Schools of Mechanical and Chemical Engineering. Associate Faculty include Professor Fred Lytle (Chemistry) Professor Galen King (Mechanical Engineering), Professor Eva Sevick (Chemical Engineering).
Cytometry Areas of Interest

Expert Systems for Diagnosis of Hematological Diseases

Although an enormous amount of data is derived from flow cytometric evaluations of patients, it is not readily analyzed. Further, without new developments in computer technology, computer programming, and analysis algorithms, it is too difficult to utilize available data in a timely manner. It can often take 2 or 3 days for complete analysis of flow cytometry data by hospital laboratories. Unfortunately, because of the complexity of the data, it is impossible to extract all of the useful information from the present data structure. New developments by the Cytometry Laboratories have enabled experts from the School of Mechanical Engineering Department of Control Systems to apply the concepts of expert systems and neural networks for rapid analysis. It should be possible to determine in minutes many of the diagnostic features from within complex multiparameter data produced on modern day flow cytometers. Associate Faculty include Professor Galen King (Mechanical Engineering), Professor Okan Ersoy (Electrical Engineering).
Cytometry Areas of Interest

Fluid Sorting and Distribution

Using well known principles of fluid switching, scientists from the School of Mechanical Engineering are attempting to develop new applications in sorting single cells in flow cytometry. Since all measurements made in flow cytometry are based upon cells moving within a stream, separating individual cells at a high sort rate and with a high degree of precision should be possible. This project may have significance to chemical engineers interested in isolating specific organisms used in biotechnology applications.
Cytometry Areas of Interest

Kinetic Measurements of Viable Human Blood Cells

When white blood cells perform their normal functions, they operate within the body under control of the immune system. Each cell type has a specific function, and measurement of these functions in vitro is difficult. Concepts envisioned by scientists from both biological and engineering disciplines would allow a replication of the in vivo environment of a blood vessel and be able to monitor many hundreds of operating blood cells within that environment. This is termed the kinetic environmental module and is a high-tech environment in which is a close replication of a closed blood vessel. Solutions can be infused or removed from the cell and cells can be added or removed. Functioning cells within the vessel can be monitored continuously as individual cells and biochemical pathways monitored. While this sounds a relatively simple task, it is enormously complex and requires many advanced technologies, including laser beam-controlled signal measurement, complex timing and signal identification, hydrodynamic systems, and computer data analysis operations.
Cytometry Areas of Interest

New Blue-Green Diode Lasers

PUCL is working with professors from the School of Electrical Engineering to apply the newly developed blue-green diode lasers to flow cytometry. This concept will allow instruments to be designed as small modules rather than very large laboratory instruments. Associate Faculty include Professors Kevin Webb, Gerald Neudack, Andrew Weiner (Electrical Engineering)
Cytometry Areas of Interest

Agricultural / Veterinary / Biomedical Interactions

There are a number of ongoing agriculture / veterinary / biomedical projects in the cytometry laboratory. (Projects menu)

Immune Dysfunction & Endotoxemia

Patients who become infected with certain micro-organisms or who undergo surgery or suffer significant burns can develop a very serious condition termed endotoxic shock. The syndrome has severe and even fatal consequences for many patients. A similar problem occurs in many farm animals, resulting in severe economic cost to the nation. Studies using flow cytometry as a major tool are being performed to try to understand what are the critical factors in endotoxic shock and how we may more readily alleviate these symptoms. Associate Faculty include Professor Geral Bottoms (Basic Medical Sciences).
Cytometry Areas of Interest

Trauma and Thermal Injury

Scientists within the facility are attempting to design a cell culture model that will replicate the symptoms of endotoxic shock resulting from trauma such as surgery, thermal injury, or severe accidents. Certain highly responsive immune cells are grown in culture and then subjected to the same biochemical stress as in a human trauma. If this program can demonstrate the importance of this modality, it may be most useful in assisting us to determine the most appropriate methods for treatment of shock. We are currently working with cultures of pulmonary artery endothelial cells which can be manipulated to identify some of the mechanisms important for oxidative tissue damage in trauma.
Cytometry Areas of Interest

Prognosis of Malignancies

Several scientists are involved in studies of cancers in an attempt to predict the course of the disease at an early stage. This would be most useful in determining treatment regimens and would be more accurate in predicting the outcome of the disease. Techniques used include measuring the exact cell cycle for the replication of malignant cells. Sometimes this information determines the type and dosage of chemotherapeutic drugs to combat the disease. These studies have application to both human and veterinary medicine and the economic impact could be pronounced in both areas.
Cytometry Areas of Interest

Determination of Tetraploidy in Cultivated Fish

Tetraploid fish are particularly useful in situations where it is not desirable to have rapidly breeding fish introduced into a new environment. Further, because these fish are sterile, they grow more rapidly and contain a higher amount of protein than regular diploid fish. The process involved in producing tetraploid fish is of interest to scientist in the School of Agriculture. A new technique has been developed with the Cytometry Laboratories to determine ploidy of these fish within a couple of days of hatching, eliminating the need to wait several weeks for the fish to grow to sufficient size. This project has other implications - it is also possible to distinguish many strains of fish very rapidly by their DNA content. Associate Faculty include Professor Chris Bidwell (Animal Science).
Cytometry Areas of Interest

Identification of Bovine Leukotoxins

Scientists from the Animal Diseases Diagnostic Laboratories together with staff from the Cytometry Laboratories are developing new ways of evaluating the effects of leukotoxins on bovine neutrophils. Using flow cytometry and multivariate analysis techniques it is possible to monitor in real time the effects of these toxins. New understandings of the mechanism of action of such toxins will result from these studies. Associate Faculty include Professor Terry Bowersock (Pathobiology).
Cytometry Areas of Interest

Sorting & Identification of Plant Chromosomes

Scientists from the Department of Horticulture are using modern laser-based flow cytometry techniques for isolating plant protoplasts. Such applications are at the leading edge in this field. The approach allows identification and physical isolation of single plant chromosomes.
Cytometry Areas of Interest

Confocal Microscopy

The facility has 2 confocal microscopes, plus a significant assortment of image analysis equipment. We are interested in using these systems for both basic image analysis as well as in new application development. Current areas of interest include development of imaging systems for detections of tissue injury, tumors, and related areas.
a. Bio-Rad MRC 1024 UV/Vis System: This system is a 2 laser system with UV lines at 353 & 361, 488 & 514 nm. In addition several other lines are available from an Ar-Kr laser (488 nm, 568 nm, & 647 nm). Simultaneous excitation of almost any combination is possible. There are 3 PMT detectors as well as a transmission detector. The system is mounted on a Nikon 300 inverted microscope, which has heated stages and objectives to assist in physiological studies.
b. DVC 250 Direct View Confocal Microscope: This system is a direct view scope mounted on a Nikon 200 inverted microscope. An intensified video camera collects signals via an image analysis computer. The system is very useful for fast fluorescence studies. For more details on the facilities go to Confocal Capabilities. Associate Faculty include Professor Kevin Webb (Electrical Engineering), Professor Eva Sevick (Chemical Engineering), Professor Galen King (Mechanical Engineering).
Cytometry Areas of Interest

Educational Activities

The Facility has a strong interest in developing teaching tools, particularly interactive computer tools.

"Handbook of Flow Cytometry Methods"

J.Paul Robinson
Zbigniew Darzynkiewicz
Phillip Dean
Lynn Dressler
Hans Tanke
Leon Wheeless

The Handbook contains 250 pages of basic and advanced flow cytometry methods. In addition, suppliers and chemicals used are listed. The Handbook is a compilation of methods from many research scientists throughtout the world, and provides some useful insights into the practical problems of running flow cytometry assays. More information can be found about the Handbook by going to [References Areas in Flow Cytometry].

The Handbook is available from:

Wiley-Liss, Inc.,
605 Third Avenue
New York, NY, 10158-0012
ISBN 0-471-59634-5 The cost is $39.95.

The current edition was published in 1993.

Cytometry Areas of Interest

STEP - Specialty Training and Education Program of ISAC

This program is currently under development. It was initiated by Dr. Zbigniew Darzynkiewicz during his Presidency of the International Society for Analytical Cytology. The purpose of the program is to provide information regarding training and educational opportunities in the field of cytometry. We now have an Introductory Homepage for STEP which you can visit. If you feel your laboratory might be able to participate as a provider of training or educational materials, please email me directly now at

Cytometry Areas of Interest

Current Protocols in Cytometry

Current Protocols in Cytometry is under development as a project between the International Society for Analytical Cytology and the publisher Wiley-Liss. The project is designed to provide up-to-date, comprehensive protocols for use with cytometry technologies such as flow cytometry and image analysis. It is possible to submit suggestions for protocols for inclusion in the volume by sending them to me at the address below or email me directly now with your suggestions:

Send an email message now to suggest protocols to J.Paul Robinson:

J.Paul Robinson, Ph.D.
Current Protocols in Cytometry Editorial Office
1515 Hansen Hall
Purdue University
West Lafayette, IN 47907-1515
Phone: (317) 494-0757 FAX (317) 494-0517

Instructions for Submission of Protocols

Instructions are under development.

Cytometry Areas of Interest

For Other exciting trips to Cytometry Environments...

Back to Purdue University Cytometry Labs

For information contact J.Paul Robinson, Ph.D., Director PUCL, 1515 Hansen B050, Purdue University, West Lafayette, IN 47907-1515, U.S.A. Phone: (317) 494-0757; Fax: (317) 494-0517 email: