Abstract

We have several thriving programs in environmental endocrinology that draw some of the best students in the nation. Many of these students are conducting hormone assays on samples that require a gamma counter. Often these samples must also be freeze dried prior to extraction. All of these students, from many labs within biology, and several departments outside biology, are sharing a single gamma counter and a single lyophilizer for this work. However, both of these instruments are old, have frequent breakdowns, and are in desperate need of replacement. We are also in need of an automated slide stage for accompanying cytological work. We are requesting a new high throughput gamma counter, a lyophilizer and an automated slide stage to accomodate these demands. This equipment will serve the needs of numerous graduate and undergraduate students from multiple departments and also help to assure that our programs maintain their world renoun graduate programs in environmental endocrinology.

Background

The Department of Biology is renown for its conservation and environmental endocrinology programs. We draw some of the best graduate students in the world to work in these areas. The labs of Professors Wingfield and Wasser measure a wide variety of hormones in response to the environment. These include studies of arctic biology, migration and molt in numerous avian species, and impacts of environmental disturbance in a wide variety of birds and mammals, from spotted owls to elephants, maned wolves, grizzly bears, elephants and sea lions. Other labs, such as that by Horacio de la Iglasia in Biology measure the endocrine basis of biological clocks. The lab of Elliott Brenowitz in Psychology and Biology focuses on environmental neuroendocrinology and the neuroendocrine basis of bird song. Several of these labs also have valuable collaborations with students in Biomedical and Health Informatics. Numerous graduate and undergraduate students are trained in these labs from multiple departments, conducting their own independent research under the guidance of our world renown faculty. Students from these programs are frequently developing new endocrine measures and applications under the guidance of these laboratories. Most of the hormone assays rely on gamma counter measurement of I125 isotopes. Students from other labs in Biology have also expressed a need for a shared gamma counter to measure other isotopes such as chromium and 55Fe. Many samples also require freeze drying prior to processing. All of our students are currently sharing the same antiquated equipment to conduct this work (a single gamma counter and lyophilizer). These essential, high use equipment items are constantly in a state of disrepair and may soon be irreparable. The aging counter has lost one of its diodes and now reads on only a single channel. Failure of this last remaining channel is imminent, but even now, the unit's slow rate of processing and limited capacity already results in project delays given the large number of student users. When the unit does finally fail, many student projects currently relying on the unit will be not only inconvenienced, but truly jeopardized. A similar case can be made for the single shared lyophilizer. In the last year alone this instrument has broken down three times, resulting in several months during which no samples could be processed, and yielding a processing backlog that furthers processing delays. These problems have impacted project timelines, deadlines for funding applications, and project reports to be filed with granting agencies and collaborators.
A new, high throughput gamma counter is desperately needed to meet the growing demands of these students across departments if these labs are to continue mentoring numerous high quality students in these ground-breaking techniques. A new lyophilizer is similarly needed to accommodate demand. The requested automated slide stage would address a different kind of technological bottleneck. In many research projects, the sheer volume of data to be processsed can create delays if the data processing resources are limited. Biomedical Health and Informatics (BHI) currently has a microscope for digital slide imaging (itself a previous STF award to BHI) that is being used for reproductive studies by students in biology and psychology. The requested automated slide stage would improve efficiency and accuracy of data analyses, greatly enhancing student productivity by resulting in an approximately 10-fold net gain in cytology slide processing time. For example: a BHI student recently adapted human cancer cell screening techniques to the question of addressing day-to-day reproductive status, via examination of Pap smear slides taken semi-daily from a variety of bears. Currently, she must view the infinitely variable cell shapes to make subjective determinations of both morphological and chromic (color) judgments for each cell counted. Thus, the larger and more comprehensive the study, the more time-consuming and problematic becomes the subjective nature of the processing. An automated slide stage would solve these problems by enabling students to create automatic routines for “recognizing” the characteristics of the cells from digitized images.

Benefits

This equipment will benefit graduate and undergraduate students conducting a wide variety of research in environmental biology. Demands for these items are quite high owing to the novel research being conducting in the department of biology and the degree to which it spans across departments such as psychology, and Biomedical Health and Informatics. Investment in the equipment outlined in this proposal will benefit the kinds of student biological research described above by facilitating project development and alleviating existing technology bottlenecks that are detrimental to ongoing research. In some cases, the equipment will reduce the time required to accomplish basic tasks-time that could be more profitably spent on the research itself. In other cases, the equipment will prevent the real possibility of permanent failure of existing equipment and the associated threat to many student research initiatives. In these ways, the requested items will enhance the research options open to students in a range of disciplines, promoting creative interdisciplinary biological research, for the current and future generations of students. Without these items, our ability to accomodate this range and diversity of high quality students could be significantly impaired.

Student Access

The freeze-dryer and gamma counter are both in areas that are secured but utilized by multiple labs. Students will need to have signed out a key and be briefed on protocols ensuring safety and consideration of shared laboratory space and equipment prior to being given unrestricted access. The university also requires that students undergo a radiation safety training course (provided free by UW) prior to using radioisotopes. The automated slide stage will be housed in the BHI building where its associated equipment from a previous STF award are housed.

Available Resources

The gamma counter and lyophilizer will complement other tools used in a variety of laboratories within and beyond the department of biology. These labs have state-of-the-art endocrine and genetic analysis facilities that include: plate readers, ß counters, refrigerated and high speed micro centrifuges, fume hoods, environmental chambers, fume hoods, column chromatography, HPLC, multiple PCR machines and automated genetic analyzers. The latter items are relevant because some laboratories measure DNA and hormones from the same samples as part of their comprehensive designs. The BHI facility includes technical support and a microscope for digital slide imaging that will be complemented by the automated slide stage.

Installation Timeline

Demands for this equipment are already quite high. Thus, all items will be put to immediate use and used heavily throughout the lifetime of the equipment.

Departmental Endorsement

The Wingfield lab uses gamma counters to count iodine-125 in radioimmunoassays for several proteins hormones (LH, FSH, Prolactin, gonaodotropin-releasing hormone, gonadotropin inhibiting hormone). We also use it to test for receptor binding in tissues. Currently I have three postdoctoral fellows and 10 graduate students who use this equipment regularly.

John Wingfield
Professor, Biology

Sam:
My students would definitely have use for the proposed gamma counter. Good luck!
Elliot Brenowitz
Professor, Departments of Psychology and Biology

Hi Sam,

My lab would potentially like to use a gamma counter.
Count us in (so to speak!)
Billie Swalla
Department of Biology
bjswalla@u.washington.edu


We will use the gamma counter for radioimmunoassays.
Horacio de la Iglasia
Department of Biology
horaciod@u.washington.edu


Sam,
I have applied for a USFWS contract to work on endangered Mariana crows and I've made it to the interview stage. If funded, this contract would support a postdoc, two grad students and three undergraduate students, and we would make use of this equipment to look at reproduction and stress.
Renee
Research Scientist
Department of Psychology

Student Endorsement

Hi Sam,

I am planning on doing work with 55Fe which is a gamma emitter. A good gamma counter would be invaluable to my thesis research.

Let me know if you'd like more details from me or my PI, Rose Ann.

cheers,

Michael Lakeman
Doctoral Candidate
Biology Department

Sam
I could definitely use the gamma counter and lyophilizer in my research.
Lisa Hayward
Doctoral Candidate
Biology Department

To whom it may concern;

My name is Sara O'Brien. I am a third year graduate student in John Wingfield's lab. I am currently studying the mechanisms controlling seasonal breeding in white-crowned sparrows. In order to look at profiles of plasma lueteinizing hormone (LH) in these birds, it is paramount that we have use of a dependable & efficient gamma counter.

We often run hundreds to thousands of samples from months & months of fieldwork. A dependable machine would allow peace of mind from having to worry about small & finite plasma samples gleaned from months of fieldwork coming to fruition. An efficient machine would allow us to count the samples more quickly, allowing us to interpret data faster & thus assess what fieldwork still needs to be done & what questions still need to be explored.

Thank you for your time and consideration,

Sara O'Brien
Graduate Student/Research Assistant
University of Washington
Department of Biology
sobrien@u.washington.edu


Sam,
A gamma counter and lyophilizer will be essential for my research on endocrinology of free-ranging elephants.
Kathleen Gobush
Doctoral Candidate
Biology

I will definitely be needing a gamma counter and lyophilizer for my Ph.D. work on impacts of environmental disturbance in maned wolves.

Carly Vynne
Biology Department


I am a student in my fourth year of graduate school in the Psychology department. My project focuses on both biology and behavior. I have utilized 4-5 undergraduate 499 students each year of the study. These undergraduates in turn have developed their own ideas and agendas related to the study of reproduction. We all rely on present and future access to the gamma counter, lyophilizer and automated slide stage tol enable our research - Cheryl Frederick.

Hi Sam-

I'm a student in Neurobiology and Behavior, but lab is based in the Psych
department. I would probably use lab equipment such a this, so feel free to
put me down as someone who's endorsed this proposal.

Thanks,

Chris Thompson


To Whom It May Concern:

As a second year graduate student in the psychology department’s animal behavior program, I plan to conduct a physiological and behavioral study of non-human primates. This work will certainly require freeze-drying and endocrine assay of fecal or salivary samples. Access to a reliable freeze drier and gamma counter will be a necessity for the successful completion of my study.

Sincerely,
Hayley Alloway

Items

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