Innovation Can Save Us
Innovation can save us. If developing interesting new technologies and products really is the lifeblood of economic health, then the life sciences industry is innovation’s beating heart.
The Scientist received more than 60 entries to our third annual Top 10 Innovations competition, presenting our judges—Northwestern University molecular chemist Neil Kelleher, sequencing pioneer Jonathan Rothberg, Princeton University genomicist Amy Caudy, and Pacific Northwest National Laboratory biologist H. Steven Wiley—with the very challenging task of winnowing these products down to the 10 best.
This year’s winners include essential tools, such as sequencers, imagers, and cell counters, that have the potential to simplify and streamline work in biology labs; and cutting-edge advances, from tailor-made disease-model cell lines to heart cells derived from induced pluripotent stem cells.
Take a look at 2010’s Top Innovations. Their clever designs speak volumes about the bright future of scientific experimentation.
* Third-Gen Sequencing
Courtesy of Pacific Biosciences
The long awaited “third-generation” sequencer from Pacific Biosciences takes first place in this year’s Top 10 Innovations contest. The technology qualifies as belonging to a new era because it’s “the first single-molecule real-time sequencer,” says Stephen Turner, the machine’s coinventor and the company’s chief technology officer, speaking to a packed auditorium at this year’s American Society of Human Genetics meeting.
Like other single-molecule sequencing machines, the PacBio RS reads a burst of fluorescent color as a tagged nucleotide is incorporated into a single molecule of DNA. However, what differentiates this technology from others is that the anchored DNA polymerase processes nucleotide binding in real time. Most second-generation technologies wash each type of nucleotide over the polymerase one at a time, simplifying detection, but slowing the process. The PacBio RS eliminates the need for multiple washings by anchoring a single DNA polymerase at the bottom of a chamber. The labeled nucleotides diffuse freely into the well, where they are excited by a laser and their fluorescence is detected by optics on the transparent underside of the chamber. Because the laser light emits a wavelength of about 600nm, it can’t penetrate farther than the bottom part of the 70-nm-wide well, keeping labeled nucleotides outside of the well in the dark, and thus greatly reducing background signal.
The surprising advantage of real-time sequencing is that the machine can detect a natural stalling when, for example, the DNA polymerase encounters a methylated or otherwise modified base. The amount of time the polymerase stalls can be used to calculate different epigenetic modifications, adding a new layer of information to the sequencing data. The instrument costs $695,000; consumables and sequencing kits are sold separately.
Rothberg: PacBio RS is a true technical tour de force. Nothing but awe comes with the observations of single molecules of DNA read out to thousands of bases—truly a seductive technology.
Caudy: Ultrafast sample analysis and long read lengths make this an exciting new entrant to the DNA sequencing field. My early-adopting pals suggest there are some initial hiccups—hope they can deliver on the promises.
* Handheld Automated Cell Counter
Courtesy of EMD Millipore
Forget the days of squinting at slides with a clicker in hand, or operating bulky benchtop machines to determine the number of cells in your sample. Cell counting is now portable, using the new Scepter Handheld Automated Cell Counter from EMD Millipore.
“It takes the tedium away,” says Grace Johnston, product manager for Scepter. Introduced in March 2010, the EMD Millipore Scepter—currently the only handheld automated cell counter available—sold over 1,000 units in its first six months on the market.
The Scepter handles like a pipette and is equipped with a screen that displays instructions to guide the user through the process. “A lot of scientists get nervous about adapting to an automated instrument, but it’s really straightforward and easy to use,” says Johnston.
Instead of relying on object recognition software like some automated benchtop counters, the Scepter draws samples into a disposable sensor where they pass through an opening charged with a current. As cells disrupt the current, the Scepter records each change in voltage. Within seconds—14, on average—the screen displays cell concentration, average cell diameter, and average cell volume, as well as histograms of each distribution.
“You can have accurate and reliable cell counts from one sample to the next, and all of that can be done right at the culture hood within 30 seconds,” says Johnston. At a list price of $2,995, it’s also the most inexpensive automated cell counter on the market, she adds.
Wiley: Cell counting is normally a very tedious process and usually only provides minimal information on the cell population. This instrument, which is only slightly larger than an automatic pipette, allows you to count cells in your tissue-culture hood, simplifies the procedure, and provides much useful data, such as the fraction of intact cells.
Caudy: At last, an alternative to lining up for the Coulter counter, and far easier than sweating over fragile hemocytometers.
* The Diffinity RapidTip
The Diffinity RapidTip is a one-step pipette tip for use in DNA purification. Following a polymerase chain reaction (PCR), samples contain more than just the DNA of interest. They also contain nucleotides, primers, and other impurities that must be removed. Traditional techniques for purifying the DNA involve several steps of washing, buffering, and rinsing that can take up to 30 minutes or longer. With the Diffinity RapidTip, all those steps are combined into a single, normal-looking pipette tip.
The product is extremely easy to use, says the company’s CEO and president Jeff Helfer. “Put [the tip] on the pipettor, aspirate, and dispense. It’s that simple.” The process requires 10–12 repetitions of pulling up and releasing the solution, and takes about one minute, making it about 50 times faster than traditional post-PCR purification techniques, Helfer says. The company plans to release a newer version of the RapidTip in January, one that would require only two or three repetition cycles, making it even more efficient.
“You start and end the [purification] process with the very same disposable pipette tip,” Helfer says. “It’s green, much less expensive, and at the end of the day, we improve lab work flow and productivity.”
The tips contain a proprietary substance that removes the impurities from a PCR reaction while simultaneously repelling the amplified double-stranded DNA of interest. Using similar differential-affinity technology, Diffinity is developing several other tips for use in a variety of applications, including automated applications, restriction-digest experiments, DNA extraction from electrophoresis gels, and next-generation sequencing library preparation. The list price is $1.50/tip, available in boxes of 48 or 96. Discounts and free samples are available.
Wiley: A great technology that saves time and effort in the lab while improving sample handling and experimental reproducibility. This would be great when using robotics.
Caudy: Finally, the convenience I’ve enjoyed for years in peptide sample cleanup, applied to DNA.
* Heart Cells on Demand
iCell Cardiomyocytes are essentially human heart cells in a test tube. Researchers at Cellular Dynamics International (CDI) induce human fibroblasts to become pluripotent stem cells (iPSC). The iPSCs are then reprogrammed to become a mixture of cells that are representative of the human heart and exhibit the typical electrophysiological characteristics of a living heart.
“The main purpose of [the] iCell Cardiomyocytes product is for drug discovery,” says Joleen Rau, senior director of marketing and communications at CDI. “Cardiotoxicity is a serious problem in drug development and is the second biggest reason for drug withdrawal from the market. We saw a market need based on a serious human health issue and realized there was an opportunity to save pharma money, make drug development safer, and perhaps save lives.”
The cardiomyocytes express monomeric red fluorescent protein, which allows for their easy identification under appropriate conditions, and a blasticidin-resistance gene, which allows CDI to achieve cardiomyocyte cultures that are at least 95 percent pure.
CDI can also create iCell Cardiomyocytes from peripheral blood samples, meaning that doctors and researchers can send in blood drawn from any human donor and have CDI generate the iPSCs needed to make personalized cardiomyocyte cultures.
“This capability to generate cells from diverse groups will help our customers to better understand how drug effects vary across different populations,” Rau says, as well as to “generate cardiomyocytes from patients afflicted with diseases such as hypertrophy and long QT syndrome (a potentially fatal condition), which will also aid in drug discovery.”
A vial that contains a minimum of 1.5 million plateable cells lists for $1,500, and is guaranteed to cover a single 96-well plate.
Kelleher: A symbol of just how fast a basic-science breakthrough can lead to new products.
Wiley: This is the first of what we expect to be many commercially available cell lines from differentiated human stem cells. This will start to move experimental biology from using the most convenient types of cell to those most relevant to a particular study
* Biologic Fluorescence Movies in Focus
Watching drugs or biologics pulse through a patient in real time usually takes expensive equipment such as a PET and/or CT scanner. For in vivo mouse studies on tight budgets researchers commonly bind a fluorescent marker to the compound of interest, and take a fluorescence snapshot. The natural background fluorescence of the entire mouse—which makes it hard to distinguish the target from normal tissue—is then subtracted away to sharpen the image. But subtracting background fluorescence in a live movie proved challenging. So the scientists and engineers at Cambridge Research & Instrumentation Inc. (CRI) took a page from PET scan technology: using the compound’s pharmacokinetics—the rate at which the drug is absorbed, circulated, and excreted—they improved the resolution by compensating for the background at every time point.
The kinetic imaging movie is of a bolus of indocyanine green travelling through the vasculature of a mouse over about 2 minutes following a tail vein injection. The dye mixes into the general blood pool, then accumulates in the liver.
The technology, called the Maestro Dynamic, could be especially useful for tracking how long cancer drugs remain at their target before being metabolized and/or excreted. Normally, to obtain data about drug accumulation in organs or tumors, one would sacrifice a cohort of mice every hour or two over the course of a day. By continually collecting data in real time, says James Mansfield, director of the company’s multispectral imaging systems, the number of mice needed could be reduced from around 100–200 to about 10.
Fluorescent labels can only yield information to a depth of a few centimeters, which just about covers the depth of the average mouse from all sides. For use in humans, however, CRI researchers have designed a mount for their fluorescence-detection camera “that you can swing over top of a surgical suite,” says Mansfield, giving doctors the ability to image the surface of the organs they’re working on in real time, to check, for example, that they’ve removed all of a tumor. The list price in the United States is $230,000.
Wiley: A kinetic in vivo imaging system that generates time-based kinetic images of fluorescent reagents and labeled antibodies. The kinetic data is used to greatly enhance the information that can be extracted from in vivo imaging, thus extending the usability of this technology to a far greater number of applications.
Caudy: The Maestro Dynamic takes whole-animal imaging from static to dynamic, operating over a range well into the tissue-permeating near-infrared spectrum.
* Centering Cells with Sound
Invented by Applied Biosystems in California, the Attune Acoustic Focusing Cytometer is the first instrument that uses ultrasound waves to position cells flowing through a cytometer into a single line before they reach a laser-based detection device. The focusing technology allows for better efficiency without sacrificing resolution and sensitivity when quantifying and/or observing cells in real time. “The sample rates are greater than 10 times faster than traditional cytometers,” says Mike Olszowy, head of flow cytometry at Life Technologies, Applied Biosystems’ mother company.
Prior to the advent of the Attune cytometer, researchers controlling the sample stream had to choose between speed of sampling and resolution quality. Now, with the help of sound waves that line up cells in the center of the sample stream, researchers can maximize speed and resolution simultaneously and adjust the flow to perform cell-by-cell analyses at the detection point. The new machine can thus allow researchers to more efficiently identify cell surface proteins expressed by cells (immunophenotyping), detect rare cell populations, quantify DNA binding to cell surfaces, or simply count cells. Currently selling for around $100,000, the device was put on the market in June 2010, and Applied Biosystems has sold more than 25 of the benchtop counters worldwide.
Wiley: Designed to use sound waves to precisely control the movement of cells and increase instrument simplicity, sensitivity and throughput. Looks like it will be particularly useful for analyzing dilute cell samples. The simplicity and relatively low cost of the instrument should also increase the number of scientists who use flow cytometry.
Caudy: With a footprint small enough to fit in a laminar-flow hood and a completely new approach to fluidics, the Attune cytometer promises less clogging than other flow cytometers, even while speeding through huge populations of cells.
* Picture-Perfect Gels
Courtesy of Bio-Rad Laboratories
Gel electrophoresis and blotting techniques are by far the most commonly employed methods for the identification and quantification of specific DNA, RNA, and proteins in a sample. But very often, capturing quality images of the separated bands and readying them for publication using photo-editing software can be laborious and time-consuming. With Gel Doc EZ, the newest gel imaging system from Bio-Rad Laboratories, researchers can load a gel and get print-quality images of up to 1200 dpi in seconds with just “a single push of a button,” says Ryan Short, marketing manager for Bio-Rad imaging.
The most user-friendly and versatile of the gel documentation systems on the market, according to Short, the Gel Doc EZ system offers four specialized gel trays for the imaging of fluorescent, colorimetric, and SYBR Green stains, as well as a novel stain-free option for imaging protein gels that circumvents the multiple washing and staining steps required. “The stain-free application can save scientists hours, if they’re doing traditional protein staining,” Short says.
With a high-quality camera and lens packed into a housing that’s just 44 x 26 x 38 cm, the system is also markedly compact and can easily fit on a benchtop with room to spare. The Gel Doc EZ costs $8,350 and includes software for image acquisition and analysis. Trays are sold separately and are priced at $1,150 each, with the exception of the stain-free tray, which costs $3,350. Stain-free precast gels are sold for $15 and $16.
Rothberg: Sometimes the best innovations are products that make the things you do simpler, faster, and cheaper. The Gel Doc EZ imager is one of those products.
Wiley: Combines a number of innovations to make a tedious lab chore easy. This is clearly a case where the whole is much greater than the sum of its parts.