New Dimensions in Confocal Microscopy

New Dimensions in Confocal Microscopy PerkinElmer's new UltraVIEW RS microscope produces real-time images in six dimensions Courtesy of PerkinElmer A protein's function depends on its movement inside a cell and interactions with other proteins and biological structures. Biologists who want to watch proteins working in living cells frequently turn to confocal microscopy. "Confocal microscopy is critical for modern cell biology and physiology," says Michael Nathanson of the Yale University School of Medicine. "Without exaggeration, it has revolutionized both of these fields, because it has allowed intracell-ular structures to be localized and intracellular events to be monitored in live cells that we have never been able to observe previously." Biologists can now observe more cellular detail than ever with the UltraVIEW RS, a new confocal microscope from Boston-based PerkinElmer Life and Analytical Sciences. MULTIDIMENSIONAL MICROSCOPY PerkinElmer advertises this new tool as "the first real-time six dimensional confocal microscope." If you wonder what six dimensions means, you are not alone. A small--admittedly unscientific--poll of confocal experts turned up no one who could name all six. Here's the answer: The UltraVIEW RS collects a sample's location in three spatial dimensions plus time. It can also scan two colors in addition to the one already covered by the first four dimensions, for a total of six. That gives researchers plenty to watch. Fedja Bobanovic, global product manager for PerkinElmer says, "UltraVIEW particularly excels for imaging green fluorescent protein and its variants. That's the real focus." He adds, "You can watch where a protein is, where it moves and what it interacts with." So far, though, Bobanovic says that very few labs routinely express three different fluorescent tags in the same cell. And, some potential users doubt the practical uses of the advertised six dimensions. Rick Rogers, director of the Biomedical Imaging Laboratory at Harvard University, says, "I take this to be more of a marketing feature, and not a practical representation of the instrument's daily use." SPIN CONTROL Still, the UltraVIEW RS, like other spinning-disk, or Nipkow disk, confocal microscopes, works faster than conventional confocals. Traditionally, confocal microscopes move laser light, point by point, in a line across a sample, rasterizing the image. While a conventional confocal microscope sends a lightning bolt of laser light down on one focused point, the UltraVIEW RS and other spinning-disk scopes shower an area with less powerful laser light by sending it through the whirling disk, which has thousands of pinholes in it. German inventor Paul Nipkow invented his disk in the 1880s as a way to transmit visual signals, but the device he conceived now plays an integral role in television and other modern electronics.1 The Molecular Expressions Web site (www.microscopy.fsu.edu) describes how it works in confocal microscopy: "Each illuminated pinhole on the spinning disk is imaged by the objective to a diffraction-limited spot on the specimen. Light reflected from the specimen can be observed in the eyepieces (or with a CCD [charge-coupled device] camera system) after it has passed back through a conjugate pinhole in the Nipkow disk. Several thousand points are simultaneously illuminated on the disk to mimic the effect of several thousand confocal microscopes running in parallel. The rapidly spinning disk fills spaces between the holes to create a real-time confocal image." Bobanovic claims that the UltraVIEW RS can be as much as an order of magnitude faster than a conventional confocal microscope with similar image quality. "PerkinElmer's UltraVIEW microscope fills an important niche--high frame rate--for certain live-cell investigations," says Rogers, who also notes a caveat: "High frame rates require significant frame averaging to reveal suitable spatial detail, unless the emission signal is very strong, say in some calcium-imaging experiments." The UltraVIEW RS also uses a new scanner that allows for computer-controlled switching of the laser wavelength to pick out different markers. Moreover, the new Nipkow scanner is sealed to keep the optics dust free, reducing maintenance. All of this comes at a basic price tag of $175,000, which is about average for confocal microscopes. Confocal microscopy likely will keep staking new claims in life science research. It already serves as the key instrument for understanding biological events in live cells, and scientists such as Rogers see a wide range of applications for the technique. "In my opinion, confocal microscopy is now a practical tool for most biological investigations," he says. To see just what the UltraVIEW RS can do, watch some of the movies at PerkinElmer's "Live Cell Imaging" site (las.perkinelmer.com/livecellimaging/home.asp). --Mike May 1. www.microscopy.fsu.edu/optics/timeline/people/nipkow.html function sendData() { document.frm.pathName.value = location.pathname; result = false if (document.frm.score[0].checked) result = true; if (document.frm.score[1].checked) result = true; if (document.frm.score[2].checked) result = true; if (document.frm.score[3].checked) result = true; if (document.frm.score[4].checked) result = true; if (!result) alert("Please select") return result; } .myradio{background-color : #94bee5} Please indicate on a 1 - 5 scale how strongly you would recommend this article to your colleagues? Not recommended 1 2 3 4 5 Highly recommended Please register your vote

New Dimensions in Confocal Microscopy

New Dimensions in Confocal Microscopy

PerkinElmer's new UltraVIEW RS microscope produces real-time images in six dimensions

Interested in reading more?

Become a Member of