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Hardware technologies and probe development in light microscopy help drive discoveries in Cell Biology: Introduction to a virtual Symposium in The Biological Bulletin.

Light microscopy in some form has become an essential part of almost every research project in the field of cellular biology. Faster and faster image acquisition times and the ability to collect more and more photons, along with the capacity to capture and store many thousands of digital images, are increasingly common requirements of many scientific studies. At the same time, scientists are seeking methodologies that shape the light that illuminates the sample and increases detection of the emitted signal. Advances in hardware development, improvements in biological labels that increase image contrast, and the ingenuity of many scientists over the years have strengthened and refined the field of light microscopy, enabling us to view more features in cells at greater resolution. Indeed, these innovations have allowed investigators to learn more about the mechanism of cellular functions and interactions in far greater detail. Even at its earliest stage of development, the microscope was instrumental in the conception of the cell theory of Matthias Schleiden and Theodor Schwann in 1838; it also provided Robert Hooke (1635-1703) with the first observations of individual cells of cork. But it was Antonie van Leeuwenhoek (1632-1723), said to have been inspired by seeing a copy of Hooke's Micrographia, who is credited with being the first to build a high-resolution, simple lens microscope to view small, living organisms, including bacteria and spermatozoa.

Later, in the 19th century, continued refinements in optics were reducing the spherical and chromatic aberrations of the compound microscopes. Improvements and discovery in the closely related field of cellular labeling added many new and enhanced tools to the arsenal of the cell biologist. The discovery of differential staining techniques that used colorimetric and newly developed fluorescent dyes, for example, provided greater contrast in biological specimens.

We set out, in this virtual symposium, "Cellular Imaging in the Biological Sciences," to highlight both past and current developments in two main aspects of microscopy that drive progress in imaging: probes that serve as labels in biological specimens, and hardware technologies that increase the capacity to capture images. The symposium opens with Stuurman and Vale's (pp. 5-13) historical overview of various camera detectors used in biological research, and include those employed in the present day. As these authors discuss, the development and adaptation of each technology enabled incremental improvements in the capture of better signal-to-noise ratios at faster rates, along with some key biological discoveries. Instrumental in these innovations were increasing computer speeds and the development of computerized control of hardware, including digital cameras.

The next pair of articles highlights the use of, and new developments in, the cutting-edge, optical technology of light sheet fluorescence microscopy (LSFM), invented in the early 1900s by R. A. Zsigmundy to image colloids. With refinements in laser illumination and detection with fast, sensitive cameras. LSFM and single-plane illumination microscopy (SPIM) have been enhanced by Ernst Stelzer's group in Germany. Several of Stelzer's lab progeny have gone on to devise still greater modifications, rendering LSFM an essential technique for capturing live-sample images with much reduced photobleaching and phototoxicity. Jan Huisken and his lab have pushed several advancements in this field. Here, in a review by Daetwyler and Huisken (pp. 14-25), several forms of fast fluorescence light sheet microscopy are described. Hari Shroff and members of his lab at NIH used an inverted, dual-arm version to implement light sheet microscopy on samples with a typical glass bottom chamber. In the article by Kumar and co-workers (pp. 26-39), the group reports on updates of this version of lightsheet microscopy that they refer to as diSPIM, or dual-view, inverted, selective plane illumination microscopy.

The most current techniques in fluorescent microscopy to delineate and visualize function in signaling pathways are explored in the next three papers. From Joshua Rappoport's group, Mazzolini et ah, (pp. 40-60) use confocal reflectance microscopy coupled with electron microscopy to quantify the uptake--and assess the functional consequences of the uptake--of a potential environmental toxin found in fuel exhaust. Anna Ye and colleagues (pp. 61-72) investigate the mechanisms involved in the proper positioning of the cleavage furrow during cytokinesis. Fluorescence tags on proteins of interest show localization. By imaging live cells during the process of DNA segregation and cytokinesis, they report, one can learn more about where the tagged proteins may be acting. The authors used a special set of fluorescence probes to measure the transfer of the emission energy of one fluorophore to the fluorophore of its related pair in a technique called the Forster resonance energy transfer (FRET). This procedure can demonstrate close proximity of protein domains suggesting the functional state of a kinase and predicting its activity. In the last article of the group. Rajendran et al. (pp. 73-84), from Mathew Tantama's lab, appraise methodologies used in imaging the key bioenergetics signaling molecule, adenosine triphosphate. The authors trace innovations in technology over time, leading up to a description of the development of specific probes that allow the imaging of ATP in single, living cells.

Fittingly, the symposium issue closes with a review by Tani and colleagues (pp. 85-95) that details the many contributions of Shinya Inoue and others to the observation of living cells using polarized light. Inoue's discoveries and overall creative influence remain an integral part of the history of the Marine Biological Laboratory (MBL), where he continues to investigate the mechanisms of cell division.

In no way is the symposium meant to be an exhaustive exploration of this varied and continuously evolving topic. But we hope that readers will enjoy the selection of entries and appreciate the range of subject matter covered by each.

LISA A. CAMERON (1,*) AND DONNA L. McPHIE (2)

(1) Light Microscopy Core Facility, Duke University and Duke University Medical Center, Durham, North Carolina 27708; and (2) Cellular Neuropsychiatry Laboratory, McLean Hospital, Harvard Medical School and Harvard University, Belmont, Massachusetts 02478

(*) To whom correspondence should be addressed. E-mail: lisa. cameron@duke.edu
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Author:Cameron, Lisa A.; McPhie, Donna L.
Publication:The Biological Bulletin
Geographic Code:1USA
Date:Aug 1, 2016
Words:977
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