Biomedical subjects

J A Squier

Publications and source records attributed to J A Squier.

5 recordsLinked to original sources

PubMed2002

Counting dendritic spines in brain tissue slices by image correlation spectroscopy analysis.

Growth of new micrometre sized projections called dendritic spines in neurones has been linked to the encoding of long-term memories in vertebrates. Numerous studies have been carried out at both the light and electron microscopy level to quantify dendritic spine densities in brain tissue in laboratory animals. Currently, such efforts using light microscopy have relied on manual counting of spines in confocal or two-photon optical slice images of tissue containing fluorescently labelled spines. This manual approach can be slow and tedious, especially for samples with high spine densities. We introduce an alternative way of performing spine counting that uses an applied image intensity threshold followed by spatial image correlation spectroscopy (ICS) analysis. We investigated the effect of particle sizes above the diffraction limit on the autocorrelation analysis as well as the influence of background fluorescence. Our results show that, for well labelled cerebellar tissue samples imaged with a signal-to-noise ratio of 5 or greater, ICS-based spine counts can be conducted with the same 15-20% precision as manual counting, but much more rapidly.

Animals↗

PubMed2001

Femtosecond Structural Dynamics in VO2 during an Ultrafast Solid-Solid Phase Transition.

Femtosecond x-ray and visible pulses were used to probe structural and electronic dynamics during an optically driven, solid-solid phase transition in VO(2). For high interband electronic excitation (approximately 5 x 10(21) cm(-3)), a subpicosecond transformation into the high-T, rutile phase of the material is observed, simultaneous with an insulator-to-metal transition. The fast time scale observed suggests that, in this regime, the structural transition may not be thermally initiated.

Journal Article↗

PubMed2000

Anharmonic lattice dynamics in germanium measured with ultrafast x-ray diffraction.

Damping of impulsively generated coherent acoustic oscillations in a femtosecond laser-heated thin germanium film is measured as a function of fluence by means of ultrafast x-ray diffraction. By simultaneously measuring picosecond strain dynamics in the film and in the unexcited silicon substrate, we separate anharmonic damping from acoustic transmission through the buried interface. The measured damping rate and its dependence on the calculated temperature of the thermal bath is consistent with estimated four-body, elastic dephasing times (T2) for 7-GHz longitudinal acoustic phonons in germanium.

Journal Article↗

PubMed2000

Two-photon image correlation spectroscopy and image cross-correlation spectroscopy.

We introduce two-photon image correlation spectroscopy (ICS) using a video rate capable multiphoton microscope. We demonstrate how video rate two-photon microscopic imaging and image correlation analysis may be combined to measure molecular transport properties over ranges typical of biomolecules in membrane environments. Using two-photon ICS, we measured diffusion coefficients as large as 10(-8) cm2 s(-1) that matched theoretical predictions for samples of fluorescent microspheres suspended in aqueous sucrose solutions. We also show the sensitivity of the method for measuring microscopic flow using analogous test samples. We demonstrate explicitly the advantages of the image correlation approach for measurement of correlation functions with high signal-to-noise in relatively short time periods and discuss situations when these methods represent improvements over non-imaging fluorescence correlation spectroscopy. We present the first demonstration of two-photon image cross-correlation spectroscopy where we simultaneously excite (via two-photon absorption) non-identical fluorophores with a single pulsed laser. We also demonstrate cellular application of two-photon ICS for measurements of slow diffusion of green fluorescent protein/adhesion receptor constructs within the basal membrane of live CHO fibroblast cells.

Animals↗

PubMedDate not supplied

Photodisruption in the human cornea as a function of laser pulse width.

BACKGROUND: We investigated the role of laser pulse width in determining fluence thresholds and efficiency for corneal photodisruption. METHODS: A laser system that delivers a wide range of pulse energies and pulse widths was used to produce ablations at pulse widths from 100 femtoseconds (fs) to 7 nanoseconds (ns). The laser-induced breakdown fluence threshold at each pulse width was determined by monitoring individual plasma emissions. Using multiple shots, the photodisruption threshold and cutting depth at each pulse width were determined histologically. RESULTS: Corneal breakdown thresholds decreased at a faster rate from 7 ns to approximately 10 picoseconds (ps), compared to further reductions in pulse width below 10 ps, where little variation was seen. Breakdown for pulse widths below 10 ps showed little intershot variability, resulting in highly reproducible fluence thresholds. Corneal tissue examined histologically showed similar fluence dependency. CONCLUSIONS: Corneal tissue photodisruption thresholds demonstrate pulse width dependence. At pulse widths less than 10 ps and with fluences near the breakdown threshold, ablations are maximally precise and efficient. These findings suggest optimal laser parameters for corneal surgery.

Cornea↗