Developing 3D SEM in a broad biological context.

When electron microscopy (EM) was introduced in the 1930s it gave scientists their first look into the nanoworld of cells. Over the last 80 years EM has vastly increased our understanding of the complex cellular structures that underlie the diverse functions that cells need to maintain life. One drawback that has been difficult to overcome was the inherent lack of volume information, mainly due to the limit on the thickness of sections that could be viewed in a transmission electron microscope (TEM). For many years scientists struggled to achieve three-dimensional (3D) EM using serial section reconstructions, TEM tomography, and scanning EM (SEM) techniques such as freeze-fracture. Although each technique yielded some special information, they required a significant amount of time and specialist expertise to obtain even a very small 3D EM dataset. Almost 20 years ago scientists began to exploit SEMs to image blocks of embedded tissues and perform serial sectioning of these tissues inside the SEM chamber. Using first focused ion beams (FIB) and subsequently robotic ultramicrotomes (serial block-face, SBF-SEM) microscopists were able to collect large volumes of 3D EM information at resolutions that could address many important biological questions, and do so in an efficient manner. We present here some examples of 3D EM taken from the many diverse specimens that have been imaged in our core facility. We propose that the next major step forward will be to efficiently correlate functional information obtained using light microscopy (LM) with 3D EM datasets to more completely investigate the important links between cell structures and their functions.

Critical issues in the focused ion beam patterning of nanometric hole matrixes on GaAs based semiconducting devices.

This work investigates the critical issues in the focused ion beam (FIB) nanopatterning of semiconducting devices. Matrixes of holes with diameter of about 150 nm were drilled by FIB on the topmost layers of a quantum dot based device. In order to study the presence of artefacts in the active region of the device, the milling parameters were investigated. A careful analysis of the ion beam effects on the structural and morphological features of the holes, mainly due to the heterogeneous composition of the layers to be milled, demonstrated that important deviations from the expected structures, in terms of size, shape and geometry of the holes, as well as layer amorphization and damage, occur.

Femtosecond near-field scanning optical microscopy.

We have developed an instrument for optically measuring carrier dynamics in thin-film materials with approximately 150 nm lateral resolution, approximately 250 fs temporal resolution and high sensitivity. This is accomplished by combining an ultrafast pump-probe laser spectroscopic technique with a near-field scanning optical microscope. A diffraction-limited pump and near-field probe configuration is used, with a novel detection system that allows for either two-colour or degenerate pump and probe photon energies, permitting greater measurement flexibility than that reported in earlier published work. The capabilities of this instrument are proven through near-field degenerate pump-probe studies of carrier dynamics in GaAs/AIGaAs single quantum well samples locally patterned by focused ion beam (FIB) implantation. We find that lateral carrier diffusion across the nanometre-scale FIB pattern plays a significant role in the decay of the excited carriers within approximately 1 microm of the implanted stripes, an effect which could not have been resolved with a far-field system.