Hopkins Imaging Initiative to host first annual conference

The Johns Hopkins University Imaging Initiative will host the first annual Imaging Conference, October 6, 2011 at the Turner Auditorium on the medical campus. The conference features afternoon lectures from various Hopkins faculty followed by a research poster session and happy hour. Anyone interested in imaging is welcome to attend.

Speakers include Elliot McVeigh, director of the Department of Biomedical Engineering; Elliot Fishman, MD, director of diagnostic imaging at body CT at Johns Hopkins Hospital; Jerry Prince, the William B. Kouwenhoven Professor of Electrical and Computer Engineering at the Whiting School of Engineering; Xingde Li, associate professor of biomedical engineering and head of the Laboratory of Biophotonics Imaging and Therapy at the Whiting School; Peter van Zijl, professor of radiology at the school of medicine and director of the F.M. Kirby Research Center for Functional Brain Imaging; and several others to be announced.

Abstracts will be accepted until Sept 6 and conference registration will be accepted until October 1. For complete information about this event and to register, go to http://imaging.jhu.edu/conferences/imaging-conference-2011

 

 

 

 

Money makes the (research) world go ‘round

Photo Illustration by Mary Spiro.

Grant money drives research, but obtaining funding can be a daunting task for those unfamiliar with the process. Wouldn’t it be nice to have someone to show you the ropes?

That’s why three postdoctoral fellows from Johns Hopkins Institute for NanoBioTechnology were asked to present a sort of crash course in how to get those almighty research dollars. The talk, given as one of INBT’s professional development seminars on July 27 to a group of graduate, undergraduate and a few high school summer research interns, covered basics, as well as some commonly overlooked issues encountered in the grant application process.

“When applying for grant funds you have to assume that everyone else also has a good idea. Your idea has to be better than great; it has to be outstanding,” Eric Balzer told attendees. Balzer is a postdoctoral fellow with professor Konstantinos Konstantopoulos in the department of Chemical and Biomolecular Engineering.

He also advised the group to avoid novice grant writing errors such as “submitting a proposal on lung cancer to an agency that only funds breast cancer research.” In other words, read the funding agency’s mission statement.

Yanique Rattigan stressed the importance of avoiding overly complex language in grant applications. “Grant reviewers often include patient representatives who are not scientists and engineers, so you have to make sure that there is a section describing the research in lay terms that they can understand,” offered Rattigan, who is conducting research in the pathology lab of professor Anirban Maitra at the Johns Hopkins School of Medicine.

Granting agencies look to fund novel research ideas, explained Daniele Gilkes. “They want to know how your work will fill in the knowledge gaps that exist in the field. You can determine this through thorough analysis of the current literature pertinent to your area of research,” added Gilkes, who works with Denis Wirtz, the Smoot Professor of Engineering in the Department of Chemical and Bimolecular Engineering.”

The group stressed the need to edit and re-edit a grant application prior to submission, and emphasized the importance of choosing the right referee to compose letters that truly support the candidates potential for independent research.

The teams’ insight into the grant application process can be found in this SlideShare slide show, click here.

Story by Mary Spiro.

 

 

 

 

 

 

 

 

 

 

 

 

Nanobio postdocs offer trusted tips on getting grant money

Photo illustration by Mary Spiro.

Three postdoctoral fellows from Johns Hopkins Institute for NanoBioTechnology will offer a one-hour crash course in how to get those research dollars; July 27, 11 a.m. Krieger 205. Free for Hopkins community.

Funding dollars make the research world go ‘round. Few know that better than postdoctoral fellows, who would be out of work without it. As part of Johns Hopkins Institute for NanoBioTechnology’s last professional development seminar of the summer, three INBT affiliated postdoctoral fellows will offer their sage advice on preparing winning research grants.

Topics to be covered on the basic aspects of grant writing include:

  • knowing when to write a grant
  • identifying funding sources
  • planning a timeline
  • how to structure a competitive proposal
  • do’s and dont’s of grant writing and planning
  • basic science writing tips for conveying ideas clearly and succinctly

This seminar will be led by Eric Balzer, postdoctoral fellow with professor Konstantinos Konstantopoulos (ChemBE); Yanique Rattigan, postdoctoral fellow with professor Anirban Maitra (Oncology/Pathology); and Daniele Gilkes, postdoctoral fellow with professor Denis Wirtz (ChemBE).

For additional information on INBT’s professional development seminar series, contact Ashanti Edwards, INBT’s Academic Program Administrator at Ashanti@jhu.edu.

 

 

 

 

‘Just add water’ to activate freeze-dried brain cancer fighting nanoparticles

A fluorescence micrograph showing brain cancer cells producing a green fluorescent protein. DNA encoded to produce the protein was delivered to the cancer cells by new freeze-dried nanoparticles produced by Johns Hopkins biomedical engineers. Image: Stephany Tzeng/JHU

Biomedical engineers and clinicians at Johns Hopkins University have developed freeze-dried nanoparticles made of a shelf-stable polymer that only need the addition of water to activate their cancer-fighting gene therapy capabilities.

Principal investigator Jordan Green, assistant professor in the department of Biomedical Engineering at the Johns Hopkins School of Medicine, led the team that fabricated the polymer-based particles measuring 80 to 150 nanometers in diameter. Each particle, which is about the size of a virus, has the ability to carry a genetic cocktail designed to produce brain cancer cell-destroying molecules. After manufacture, the nanoparticles can be stored for up to 90 days before use. In principle, cancer therapies based on this technology could lead to a convenient commercial product that clinicians simply activate with water before injection into brain cancer tumor sites.

Because this method avoids the common, unpleasant side effects of traditional chemotherapy, “nanoparticle-based gene therapy has the potential to be both safer and more effective than conventional chemical therapies for the treatment of cancer,” Green said. But, he added current gene therapy nanoparticle preparations are just not practical for clinical use.

“A challenge in the field is that most non-viral gene therapy methods have very low efficacy. Another challenge with biodegradable nanoparticles, like the ones used here is that particle preparation typically takes multiple time-sensitive steps.” Green said. “Delay with formulation results in polymer degradation, and there can be variability between batches. Although this is a simple procedure for lab experiments, a clinician who wishes to use these particles during neurosurgery will face factors that would make the results unpredictable.”

In contrast, the nanoparticles developed by the Green lab are a freeze-dried, or “lyophilized,” formulation. “A clinician would simply add water, and it is ready to inject,” Green said. Green thinks this freeze-dried gene-delivery nanoparticle could be easily manufactured on a large scale.

Co-investigator Alfredo Quinones-Hinojosa, a Johns Hopkins Hospital clinician-scientist and associate professor in the departments of Neurosurgery and Oncology at the Johns Hopkins School of Medicine, said he could imagine particles based on this technology being used in conjunction with, and even instead of, brain surgery. “I envision that one day, as we understand the etiology and progression of brain cancer, we will be able to use these nanoparticles even before doing surgery,” Quinones said. “How nice would that be? Imagine avoiding brain surgery all together!”

Currently, patients with glioblastoma, or brain cancer, only have a median survival of about 14 months, Green said. “Methods other than the traditional chemotherapy drugs and radiation—or in combination with them—may improve prognosis,” he said.

Gene therapy approaches could also be personalized, Green said. “Because gene therapy can take advantage of many naturally-existing pathways and can be targeted to the cancer type of choice through nanoparticle design and transcriptional control, several levels of treatment specificity could be provided,” Green said.

The nanoparticles self-assemble from a polymer structural unit, so fabrication is fairly simple, said Green. Finding the right polymer to use, however, proved to be a challenge. Lead author Stephany Tzeng, a PhD student in biomedical engineering in Green’s lab screened an assortment of formulations from a “polymer library” before hitting on a winning combination.

“One challenge with a polymer library approach is that there are many polymers to be synthesized and nanoparticle formulations to be tested. Another challenge is designing the experiments to find out why the lead formulation works so well compared to other similar polymers and to commercially available reagents,” Green said.

Tzeng settled on a particular formulation of poly(beta-amino ester)s specifically attracted to glioblastoma (GB) cells and to brain tumor stem cells (BTSC), the cells responsible for tumor growth and spread. “Poly(beta-amino ester) nanoparticles are generally able to transfect many types of cells, but some are more specific to GBs and BTSCs,” Tzeng said.

The nanoparticles work like a virus, co-opting the cell’s own protein-making machinery, but in this case, to produce a reporter gene (used to delineate a tumor’s location) or new cancer fighting molecule. “It is possible that glioblastoma-derived cells, especially brain tumor stem cells, are more susceptible to our gene delivery approach because they divide much faster,” Tzeng added.

Not only are the particles convenient to use, the team discovered that dividing cells continued to make the new protein for as long as six weeks after application. “The gene expression peaked within a few days, which would correspond to a large initial dose of a therapeutic protein,” said Green. “The fact that gene expression can continue at a low level for a long time following injection could potentially cause a sustained, local delivery of the therapeutic protein without requiring subsequent injection or administration. The cells themselves would act as a ‘factory’ for the drug.”

Once the nanoparticles release their DNA cargo, Tzeng said the polymer quickly degrades in water, usually within days. “From there, we believe the degradation products are processed and excreted with other cellular waste products,” Tzeng said.

Members of the Green Lab are now working on identifying the intracellular mechanism responsible for facilitating cell-specific delivery. “We also plan to build additional levels of targeting into this system to make it even more specific. This includes modifying the nanoparticles with ligands to specifically bind to glioblastoma cells, making the DNA cargo able to be expressed only in GB cells, and using a DNA sequence whose product is only effective in GB cells.”

So far, the team has only successfully transfected brain tumor stem cells using these nanoparticles in a plastic dish. The next step is to test the particle in animal models.

“We hope to begin tests in vivo in the near future by implanting brain tumor stem cells into a mouse and injecting particles. We also hope to begin using functional genes that would kill cancer cells in addition to the fluorescent proteins that serve only as a marker,” Tzeng said.

Other authors who contributed to this work are Hugo Guerrero-Cázares, postdoctoral fellow in Neurosurgery and Oncology, and Joel Sunshine, an M.D.-Ph.D. candidate, and Elliott Martinez, an undergraduate leadership alliance summer student, both from Biomedical Engineering. Funding for this work came from the National Institutes of Health, Howard Hughes Medical Institute, the Robert Wood Johnson Foundation and a pilot-grant from Johns Hopkins Institute for NanoBioTechnology (INBT). Green is an affiliated faculty member of INBT. The research will be published in Issue #23 (August 2011) of the journal Biomaterials and is currently available online.

Freeze-dried gene therapy system avoids virus, complications

Story by Mary Spiro

 

Becton Dickinson leader to discuss medical device development

Adam Steel (Becton Dickinson)

INBT hosts a talk on medical device development from Becton Dickinson systems integration director Adam Steel, July 13, 11 a.m. in Krieger 205. Free to Hopkins community.

Adam Steel, PhD, Director of Systems Engineering at Becton Dickinson, will discuss medical device development as part of Johns Hopkins Institute for NanoBioTechnology’s professional development seminars, Wednesday July 13 at 11 a.m. in Krieger 205.

Dr. Steel joined BD in 2005. Previously he was vice president of research and development at MetriGenix. He earned his PhD in analytical chemistry at the University of Maryland College Park and undergraduate degrees in chemistry and mathematics from Gettysburg College. He completed a postdoctoral fellowship in medical device development at the National Institutes of Standards and Technology.

This talk is the third installment in INBT’s free, summer professional development seminar series. Topics are geared toward undergraduate and graduate students.

The final seminar will be held July 27 on the topic of the grant submission process and how to obtain funding for research. For additional information on INBT’s professional development seminar series, contact Ashanti Edwards, INBT’s Academic Program Administrator at Ashanti@jhu.edu.

 

 

Collagen video scores high in magazines reader’s choice vote

Screen capture from INBT’s video on collagen mimetic peptides.

The Scientist magazine has announced its annual Multimedia Awards—the Labbys—and Johns Hopkins Institute for NanoBioTechnology’s video on collagen mimetic peptides has been selected as a finalist. According to the voting, we are a strong second in the race. It appears voting is continuing well past the original June 30 deadline. So keep voting!

Help choose us as the top science video by going to this website (http://the-scientist.com/2011/06/15/2011-labby-video-finalists/#vote)  and selecting “Mimicking Collagen.” The video features Michael Yu, associate professor of materials science and engineering and some fantastic animations and illustration from INBT’s Animation studio. Animations in the video were created by Ella McCrea, a graduate from the Maryland Institute College of Art, and Nathan Weiss, a masters graduate from Johns Hopkins University.

Winners of the reader’s choice will be announced in the magazine and online in September. Top picks will also be chosen by The Scientist’s panel of judges, which includes the father of the infographic Nigel Holmes, Kirsten Sanford of the Science Channel (aka Dr. KiKi), Jeffrey Segall of the Albert Einstein College of Medicine in New York City, and David Kirby of the University of Manchester.

You can only vote once, so share this link with your friends.

 

 

Summer interns join PS-OC labs

Each summer, Johns Hopkins Institute for Nanobiotechnology (INBT) hosts several summer research interns, five of who will conduct research as part of Johns Hopkins Physical Sciences-Oncology Center.

Erin Heim, from University of Florida, will be testing the effects of cell geometry and chemotaxis on cell polarity in the Denis Wirtz lab (Chemical and Biomolecular Engineering). “The goal is to find which of the two is more important to polarity when working against each other,” she said.

Also in the Wirtz lab, Nick Trenton is developing an agarose-based microfluidics chamber that can be used to establish a chemotaxis gradient in 3D cell culture. “We’ll be testing various cell knockdowns in 3D in the presence of a chemokine gradient,” he said.

Rachel Louie from Johns Hopkins, works in the Peter Searson lab (Materials Science and Engineering). She is characterizing the properties of human umbilical vein endothelial cells cultured under different conditions. “We’re testing to see how the amount of growth factors in cell culture medium will affect transendothelial electrical resistance values,” Louie said.

Thea Roper from North Carolina State University works in the Sharon Gerecht lab (Chemical and Biomolecular Engineering). Roper said she will analyze how human embryonic stem cells mature into smooth muscle cells. “To do this, I must determine the pathway by using techniques such as immunofluorescence, RT-PCR, and Western Blot to examine Myocardin, a transcriptional co-activator, Elk-1, a ternary complex factor, PDGF-R, platelet-derived growth factor receptors, and SRF, serum response factors,” she said.

Quinton Smith also works in the Gerecht lab. This is his second year interning at Hopkins. Smith, from University of New Mexico, is fabricating a microfluidic device that recreates hypoxic (low oxygen) conditions. “I’ll study how adult and embryonic stem cells respond to this dynamic environment,” he said.

Read more about INBT’s summer interns at the following link: http://wp.me/p1sSPo-VT

 

Come to the NanoBio Film Festival 11 a.m., 6/29 in Krieger 205

Charli Dvoracek storyboarding a video. Photo by Mary Spiro

Johns Hopkins Institute for NanoBioTechnology (INBT) hosts the NanoBio Film Festival on June 29, 11 a.m. in Krieger 205. See the world premiere of three short videos made by members of INBT’s course on science communications. Free for Hopkins community.

Videos featured in this film festival describe the current research of students working in INBT affiliated laboratories. Students in the course learn how to communicate their work in nontechnical terms for general audiences. They work in teams to write, direct, film and produce the videos within a two-week time frame. The producers will be on hand to describe their experience making the videos and to answer questions.

The INBT film festival is part of the institute’s free professional development seminar series. Topics are geared toward undergraduate and graduate students.

Future seminars include:

  • July 13: Adam Steel, PhD, Director of Systems Engineering at Becton Dickinson, will discuss medical device development. Dr. Steel joined BD in 2005. Previously he was vice president of research and development at MetriGenix. He earned his PhD in analytical chemistry at the University of Maryland College Park and undergraduate degrees in chemistry and mathematics from Gettysburg College. He completed a postdoctoral fellowship in medical device development at the National Institutes of Standards and Technology.
  • July 27: Grant submission process and how to obtain funding; a roundtable discussion with INBT affiliated postdoctoral students.

For additional information on INBT’s professional development seminar series, contact Ashanti Edwards, INBT’s Academic Program Administrator at Ashanti@jhu.edu.

 

 

Johns Hopkins Integrated Imaging Center focuses on data

Shyenne Yang positions Drosophila embryos for fluorescence imaging. Photo by Marty Katz/baltimorephotographer.com

Heavy, black curtains and dimmed lights shroud the core of the Johns Hopkins Integrated Imaging Center (IIC). Yet researchers who peer through the advanced microscopes cloaked by these dark draperies view experimental samples more clearly than ever thanks to a combination of the high-tech equipment and the creative expertise offered by the center’s seven-member staff.

When describing Johns Hopkins University’s showpiece microscopy facility, it’s easy to rattle off a laundry list of available equipment and laboratory space able to prepare samples with nearly any contrasting agent found in the literature. The Homewood-based center contains devices that can image a sample in virtually any manner in 2-D, 3-D and even 4-D. IIC’s 3,500 square-foot facility comprising space in Dunning, Jenkins, and Olin Halls, boasts more than $7.5 million worth of state- of-the-art imaging equipment, including a Zeiss laser scanning microscope (LSM) 510 VIS confocal with a Confocor 3 fluorescence correlation spectroscopy (FCS) module—one of only a very few such uniquely configured laser scanning microscopes in the United States.

Director J. Michael McCaffery, a research professor in the Department of Biology at the Krieger School of Arts and Sciences, said the Hopkins community is thrilled to have access to such a versatile microscope with fluorescence correlation spectroscopy that is capable of cross-correlation analysis, with confocal imaging and a fully enclosed environmental system for live imaging. Researchers affiliated with Johns Hopkins Institute for NanoBioTechnology (INBT), the Johns Hopkins Physical Sciences Oncology Center and Center of Cancer Nanotechnology Excellence are also glad to have access to IIC’s menu of facilities.

“Fluorescence correlation spectroscopy allows for high-resolution spatial and temporal analysis of single biomolecules with respect to diffusion, binding, as well as enzymatic reactions in vitro and in vivo,” McCaffery said. In other words, you can see and measure a lot of really tiny stuff with it, something INBT affiliated researchers working at micron/nanometer resolutions are finding incredibly useful.

The center features multiple suites devoted to specific microscopy/imaging functions, as well as facilities for all manner of sample preparation. All these advanced tools help scientists and engineers characterize nanomaterials; and image cells, sub-cellular organelles, and biomolecules/ proteins at very small dimensions. But none of this fancy equipment would be of much use to researchers without the expertise of McCaffery and the IIC staff. McCaffery brings years of experience and a background in cell biology and microbiology. The center’s associate director, William Wilson, an associate research professor in the Department of Materials Science and Engineering at the Whiting School of Engineering, describes himself as a “chemist, turned physicist, who became an electrical engineer, who is now a materials scientist.”

Staff scientist Kenneth J.T. Livi, director of the IIC’s High-Resolution Analytical Electron Microbeam Facility located in Olin Hall, offers his unique perspective on earth and planetary sciences. Researchers can also consult with microscopy specialist/ trained biologist and FACS supervisor Erin Pryce, the FACS manager Yorke Zhang, computer/IT specialist Marcus Sanchez, and research assistants Leah Kim and Adrian Cotarelo, who both are currently earning their bachelor degrees in biology at Johns Hopkins.

From left, IIC director Michael McCaffery, FACS supervisor Erin Pryce, and associate director William Wilson with the BD FACSVantage SE. Photo by Mary Spiro

“Sometimes young researchers haven’t contemplated all the possibilities of how to use and apply an instrument; and don’t realize there are many different ways to utilize familiar tools in order to obtain new, in some cases better, information,” McCaffery said. “Our desire is always to approach a problem from many disparate perspectives to generate convergent data that corroborates each particular assay. Hopefully, results from each individual assay, allows the scientist to arrive at a convergent perspective that yields confidence in the results and conclusions.”

One of the easiest ways to obtain different microscopy data and improve corroboration among assays is simply to change the contrast mechanism.

“The most common contrast mechanisms used to image something are optical contrast (transparent versus opaque), polarization, and fluorescence,” said Wilson. “But there are many different ways you can manipulate how light interacts with the specimen and what you detect out of an objective.”

For example, ultrafast laser sources have made nonlinear optical forms of contrast an exciting new tool. Techniques like two-photon excited fluorescence and second harmonic generation (both available in the IIC) produce excellent spectral and structural information about samples because a smaller effective photon volume is excited. Wilson explained it like this: “Imagine turning your stereo all the way up and hearing the sound distorted. That distortion is created by the higher order acoustic harmonics from your stereo. The same happens with intense laser light resulting in new “colors” being generated from the object irradiated. The cool thing is that the different non-linear processes are often sensitive to different physical proper- ties or structural features, offering complementary information about your sample.”

In some cases, getting more detailed information simply requires looking at the right color range. The two-photon fluorescence and second harmonic signals appear at different wavelengths. If you excite a sample with enough energy to generate third order harmonics, that signal is detected at an even bluer wavelength, Wilson said. “With third harmonic generation, you only get signals from the interface of structures with no interference from anything else. This means you can simultaneously image fluorescence, polar order, and interface dynamics just by popping in a few filters and beam splitters,” he said.

“Over the past ten or so years, physicists and engineers focused on advanced microscopy, have produced better and more advanced laser and optical technologies, generating techniques that many researchers in the biological and biomedical sciences might not know exist,” Wilson said. “There also are a lot of applied physicists who are developing and using these new technologies who don’t know what an interesting sample is. We hope to help bridge this gap, becoming a place where these collaborative synergies can flourish.”

Sample preparation is another area where the center can help researchers. “Cell fractionation, for example, which is the breaking down of whole cells and separating them into their individual components, when combined with biochemical techniques and microscopy, can often allow researchers to pose more precise questions and to better analyze a biological problem,” McCaffery said.

“It is common for someone to come in and want to use a particular instrument or technique they read about in a paper,” McCaffery said. When that happens, McCaffery and Wilson are likely to give researchers “homework.”

“It’s important to remember that the goal is not to make a pretty picture,” Wilson said. “The goal is to answer a question, so sometimes we have to ask them, ‘What is your research question?’” An enviable set of microscopy tools combined with a team that brings years of training and experience from a variety of disciplines sets Johns Hopkins Integrated Imaging Center apart from the microscope on the individual researcher’s lab bench, as well as from facilities nationwide. Wherever possible, McCaffery said, IIC staff tries to be engaged in all of the research that is carried out in the center. “Simply, our involvement leads to better results and better science,” McCaffery added.

Researchers confirm this successful combination.

“The facilities at the IIC have allowed us to obtain critical information about the internal structure of our peptide nanomaterials that would have remained unknown without careful electron and fluorescence microscopy,” said J.D. Tovar, assistant professor of Chemistry. “Equally important, the scientific IIC staff members were vital participants making sure collaborative experiments were done meaningfully and students were trained competently. Our collaboration with Dr. Wilson has given some nice insights and at the same time has posed many more questions for future research.”

Praise like that for the IIC is always nice to hear, staff members say, but they emphasize that the services and tools they provide are just part of the job. “Part of being a scientist is learning not only how to gather information from a wide variety of tools but also understanding how to pose clear questions that lead to the right tools, in a nutshell, how to not waste time. If we can help you do that, then we have achieved our goal,” Wilson said.
This story originally appeared in Johns Hopkins Nano-Bio Magazine.

To read more about IIC’s facilities and services, go here.

Story by Mary Spiro

Photos by Mary Spiro and Marty Katz

 

Voting for Johns Hopkins video in multimedia contest ends June 30

The Scientist magazine has announced its annual Multimedia Awards—the Labbies—and Johns Hopkins Institute for NanoBioTechnology’s video on collagen mimetic peptides has been selected as a finalist. But that just means we are in the finals. We need your vote to win!

Help choose us as the top science video by going to this website (http://ht.ly/5mZ9D) and voting for “Mimicking Collagen.” The video features Michael Yu, associate professor of materials science and engineering and some fantastic animations and illustration from INBT’s Animation studio.

Voting ends June 30, 2011 and winners of the reader’s choice will be announced in the magazine and online in September. Top picks will also be chosen by The Scientist’s panel of judges, which includes the father of the infographic Nigel Holmes, Kirsten Sanford of the Science Channel (aka Dr. KiKi), Jeffrey Segall of the Albert Einstein College of Medicine in New York City, and David Kirby of the University of Manchester.

So vote now and often! Share this link with your friends.

Watch the video that made the Labby finals.

Collagen Mimetic Peptides