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Journal of Microscopy

An international and interdisciplinary forum for publication, discussion & education for scientists and technologists who use any form of microscopy or image analysis.

Journal_Aug 2014.jpgThe Journal of Microscopy publishes quality original research articles, review articles, Hot Topic papers, and letters to the Editor, covering all aspects of microscopy and analysis. This includes cutting-edge technology and innovative applications in physics, chemistry, material and biological sciences.

The Journal is for scientists and technologists who develop, advance or apply any form of microscopy, spatially resolved spectroscopy, compositional mapping & image analysis.

The Journal of Microscopy is particularly interested in papers on original applications & developments in microscopy.


Impact Factor: 2.15

ISI Journal Citation Reports © Ranking: 2013: 3/11 (Microscopy)
Frequency: Monthly
Print ISSN: 0022-2720 / Online ISSN: 1365-2818

Aims & Scope

The Journal of Microscopy is for scientists and technologists that use any form of microscopy, spatially resolved spectroscopy, compositional mapping and image analysis. This includes technology and applications in physics, chemistry, material and biological sciences.

The Journal of Microscopy is published by Wiley on the Wiley Online Library on behalf of the Royal Microscopical Society.

Latest Early View Papers available online

The following early view papers are available now at Wiley online.

A series of flexible design adaptations to the Nikon E-C1 and E-C2 confocal microscope systems for UV, multiphoton and FLIM imaging
Stanley W. Botchway, Kathrin M. Scherer, Steve Hook, Christopher D. Stubbs, Eleanor Weston, Roger H. Bisby and Anthony W. Parker

Cells contain proteins and compounds that may be visualized using ultra-violet (UV) excitation. Generation of fluorescence from these chromophores requires excitation in the UV. However most commercial microscopes do not have UV transmitting optics for high resolution imaging. Multiphoton excitation is a method providing the equivalent of UV excitation beneath UV absorbing surface materials permitting direct excitation of simple molecular systems that absorb below 350 nm. Multiphoton microscopy in combination with Fluorescence Lifetime Imaging Microscopy (FLIM) is now a key technique in biomedical and life-science research for live cell and whole animal imaging providing several advantages over the one photon visible confocal technique. The advantages include the use of near infra-red light leading to reduced phototoxicity and improved tissue transparency. The added benefit and advantage of FLIM over standard steady state fluorescence microscopy is that the measured lifetime of a chromophore is independent of concentration below ca. 1 mM and may be used as a reliable probe for distance-depended processes involving resonance energy transfer and also reports directly on the chemical environment of the chromophore.

Here we report a straightforward adaptation to a commercial Nikon confocal scanning system and microscope to allow simultaneous one-photon, steady state multiphoton imaging, FLIM and deep UV microscopy below 300 nm. We show that aromatic amino acid compounds and UV absorbing anticancer drugs can be detected and imaged in mammalian cells using these adaptations. The wider spectral working region has significant potential for drug studies in cells where the main excitation and emission is in the UV region of the electro-magnetic spectrum.


3D imaging of cement-based materials at submicron resolution by combining laser scanning confocal microscopy with serial sectioning
M.H.N. Yio, M.J. Mac, H.S. Wong and N.R. Buenfeld

Current technology in 3D imaging is limited by the compromise between achievable resolution and image volume. Existing techniques can only perform either high resolution imaging of small sample sizes or low resolution imaging of larger samples. In this study, we developed a new method for imaging large volumes of sample at sub-micron resolutions. Our method combines fluorescence confocal microscopy with serial sectioning. After each sectioning, we image a thin volume which overlaps with the previous one. By repeating the process, we are able to build up a large 3D image based on the overlapping features. We have demonstrated the method on cement-based materials which contain features such as pores and cracks ranging from sub-micron to hundreds of micrometres. The 3D images of these features can be used to understand and model the transport of water and gas in concrete. This will help in the development of more durable concrete structures.


Effect of the sample annealing temperature and sample crystallographic orientation on the charge kinetics of MgO single crystals subjected to keV electron irradiation
A. Boughariou, G. Damamme and A. Kallel

The present paper focuses on the effect of sample annealing temperature and crystallographic orientation on the secondary electron yield of MgO during the charge by a defocused electron beam irradiation. The experimental results show that there are two regimes during the charging process which are better identified by plotting the logarithm of the secondary electron emission yield lns as function of the total trapped charge in the material QT. The impact of the annealing temperature and crystallographic orientation on the evolution of lnσ is presented. The slope of the asymptotic regime of the curve lnσ as function of QT, expressed in cm2/ trapped charge, is probably linked to the elementary cross section of electron-hole recombination,σhole, which controls the trapping evolution in the reach of the stationary flow regime.
In this work, we have investigated the surface properties of MgO (100), (110) and (111) surface orientations, irradiated with 1.1 and 5 keV electrons by using a SEM LEO 440. We tried to characterize the charging properties of MgO by studying the variation of lnσ (logarithm of secondary electron emission yield) as function of the trapped charge QT within the material. For both crystallographic surface directions (100) and (111) the experimental results show that annealing temperature affects the logarithm of the initial yield of MgO. For the (100) surface orientation, we have lnσ0(1000°C)>lnσ0(1500°C). However, for the (111) orientation the phenomenon is reversed ie lnσ0(1000°C)<lnσ0(1500°C). In addition, two different kinds of behavior appear in the beginning of the variation of lnσ as a function of the trapped charge QT:

  • At E0=5 keV a plateau appears,
  • Yet at E0=1.1 keV a plateau does not exist.

According to the different curves we note that the slopes are the same P=1.6 10-10 cm2/charge, whatever the crystallographic orientation, the primary beam energy E0 and the annealing temperature. We can then say that the slope is completely intrinsic to the material as it is independent on initial conditions (λabs0) and condition of irradiation (lsource and Rp); it is interpreted as proportional to the recombination cross section (σhole) between free electrons and holes.


Contact X-ray microscopy of living cells by using LiF crystal as imaging detector
L. Reale, F. Bonfigli, A. Lai, F. Flora, P. Albertano, M. L. DI Giorgio, L. Mezi, R.M. Montereali, A. Faenov, T. Pikuz, S. Almaviva, M. Francucci, P. Gaudio, S. Martellucci, M. Richetta and A. Poma

The possibility of detecting images by X-ray of materials, devices and biological samples with high spatial resolution is a topical task nowadays. The great and fast development of radiation sources, optics and detectors, makes X-ray imaging applications very attractive in physics as well as in life sciences. For biological investigations, when compared with traditional microscopy techniques, single-exposure X-ray microscopy has the potential to obtain high spatial resolution images while still keeping a sample alive. In particular, in the (2.2-4.4) nm wavelength interval, known as water-window, water is much less absorbing than organic compounds. Thus images of specimens containing water can be obtained with a natural contrast. X-ray contact microscopy represents the simplest X imaging technique: the sample is placed in direct contact with a sensitive material and exposed to radiation. The image is formed in an absorption contrast process. This process doesn’t need coherent radiation and compact polychromatic laboratory X-ray sources, such as soft X-ray laser plasma sources, can be used. Single-shot experiments are possible in this experimental configuration, allowing in vivo investigation (without the need of any cryogenic or chemical process). The performances of lithium fluoride (LiF) detectors have renewed the interest in such imaging technique and in the development of X-ray imaging laboratory tools.
We present the use of LiF as imaging radiation detector to analyze living cells by single-shot soft X-ray contact microscopy. High resolved X-ray images on LiF of cyanobacterium Leptolyngbya VRUC135, two unicellular microalgae of the genus Chlamydomonas and mouse macrophage cells (line RAW 264.7) have been obtained utilizing X-ray radiation in the water window energy range from a laser plasma source. The used method is based on loading of the samples, the cell suspension, in a special holder where they are in close contact with a LiF crystal solid-state X-ray imaging detector. After exposure and sample removal, the images stored in LiF are read by an optical microscope in fluorescence mode. We showed that the peculiarities of LiF detector (wide dynamic range and very high spatial resolution) are particularly suitable for single-shot soft X-ray contact microscopy of in vivo biological samples. In particular, images revealing the internal structures of the investigated samples at high spatial resolution have been obtained. Furthermore, an image reconstruction technique for stray light removal to enhance the contrast and reveal details has been shown.

Latest Short Invited Reviews available online

Techniques for RNA in vivo imaging in plants
Jens Tilsner

Electron microscopy of quantum dots
Thomas Walther

What environmental transmission electron microscopy measures and how this links to diffusivity: thermodynamics versus kinetics
Thomas Walther

Wavelets: on the virtues and applications of the mathematical microscope
Michael Unser

Biological applications of cryo-soft X-ray tomography
E. Duke, K. Dent, M. Razi and L. M. Collinson

Nonlinear optical microscopy with vibrational contrast
M. J. Winterhalder and A. Zumbusch

Focused ion beam scanning electron microscopy in biology
C. Kizilyaprak, J. Daraspe and B.M. Humbel

Lay Descriptions

In order to make the science we publish more accessible, a second abstract or lay description in easy to understand non-technical language has been developed. Lay Descriptions for papers published through the Journal of Microsopy will be included in infocus and here on the RMS website.

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