Label-free multimodal imaging with simultaneous two-photon and three-photon microscopy and kernel-based nonlinear scaling denoising.

We report on a compact multimodal imaging system that can acquire two-photon microscopy (2PM) and three-photon microscopy (3PM) images simultaneously. With dual excitation wavelengths, multiple contrasts including two-photon-excitation-fluorescence (2PEF), second harmonic generation (SHG), and third harmonic generation (THG) are acquired simultaneously from cells, collagen fibers, and interfaces, all label-free. Challenges related to the excitation by two wavelengths and the effective separation of 2PM and 3PM signals are discussed and addressed. The data processing challenge where multiple contrasts can have significantly varying signal levels is also addressed. A kernel-based nonlinear scaling (KNS) denoising method is introduced to reduce noise from ultra-low signal images and generate high-quality multimodal images. Simultaneous 2PM and 3PM imaging is demonstrated on various tissue samples. The simultaneous acquisition speeds up the imaging process and minimizes the commonly encountered problem of motion artifacts and mechanical drift in sequential acquisition. Multimodal imaging with simultaneous 2PM and 3PM will have great potential for label-free in-vivo imaging of biological tissues.

Dual-wavelength multimodal multiphoton microscope with SMA-based depth scanning.

We report on a multimodal multiphoton microscopy (MPM) system with depth scanning. The multimodal capability is realized by an Er-doped femtosecond fiber laser with dual output wavelengths of 1580 nm and 790 nm that are responsible for three-photon and two-photon excitation, respectively. A shape-memory-alloy (SMA) actuated miniaturized objective enables the depth scanning capability. Image stacks combined with two-photon excitation fluorescence (TPEF), second harmonic generation (SHG), and third harmonic generation (THG) signals have been acquired from animal, fungus, and plant tissue samples with a maximum depth range over 200 µm.

Preliminary Clinical Results and Mechanical Behavior of a New Double-Layer Carotid Stent.

PURPOSE: To evaluate the in vitro mechanical and clinical implant behavior of a next-generation double-layer stent designed for the carotid artery. METHODS: The new double-layer CASPER-RX stent was implanted in 12 patients (median age 69 years; 8 men) with high-grade symptomatic internal carotid artery stenoses (mean 82%). In the in vitro experiments, the CASPER-RX stent (8-×40-mm model) was investigated with respect to its radial force on expansion and the bending stiffness of the stent system and of the stent in its expanded state, as well as the collapse pressure in a thin, flexible sleeve. The wall adaptation of the expanded stents was assessed by fluoroscopy after release in a step and curve model. RESULTS: Technical success was achieved in all patients without complications; there was no peri- or postinterventional stroke and no stroke or restenosis after 6 months. In the experimental studies, the bending stiffness of the stent on the delivery system (154.9 N mm(2)) was significantly lower than when expanded in a 7-mm flexible tube (467.4 N mm(2)). The radial force on expansion of the stent to 7 mm was low (0.011 N/mm). The collapse pressure was relatively high (0.56 bar) as a result of the stent's particular stent structure. The stent exhibited significant foreshortening of 27.6%. The conformability to the wall in the step model was relatively smooth; in the curve model, straightening occurred with consecutive slight stenosis. CONCLUSION: The first clinical results showed a safe implantation behavior without the occurrence of any ischemia. The structure of the new CASPER-RX stent creates an acceptable flexibility, low radial force, and high collapse pressure. The large foreshortening during implantation should be considered as well as the higher bending stiffness, especially when used in elongated carotid arteries.

A high ratio of ADP-TRAP induced platelet aggregation is associated more strongly with increased mortality after coronary stent implantation than high conventional ADP induced aggregation alone.

OBJECTIVE: This study aimed to evaluate whether a high relative ADP induced aggregation (r-ADP-agg) is associated with an increased mortality in patients after coronary stent implantation. BACKGROUND: Several trials were not able to improve clinical outcome by adapting platelet inhibition in patients after coronary stent implantation and high platelet reactivity (HPR). Platelet monitoring is complex and conventional definition of adenosindiphosphate (ADP) induced aggregation alone might not transfer the whole picture of adequate platelet inhibition in vivo. METHODS: In a prospective single-centre observational trial multiple electrode aggregometry was performed in whole blood of patients after stent implantation. r-ADP-agg was defined as the ADP-thrombin receptor activating peptide ratio to reflect an individual degree of P2Y12 dependent platelet inhibition with a cut-off value for HPR of ≥ 50%. The primary end point was mortality. RESULTS: Follow-up was completed in 176 of 184 patients (96%) with a mean follow-up time of 3.7 years. 35 (20 %) patients revealed an r-ADP-agg ≥ 50%. An r-ADP-agg ≥ 50% was associated with an increased mortality [unadjusted hazard ratio (HR) 7.006 (2.561-19.17); p = 0.0001]. In a multivariable Cox regression analysis mortality was independently associated with an r-ADP-agg ≥ 50% [HR 3.324 (1.542-7.165); p = 0.0022], ACS-setting [HR 3.249 (1.322-7.989); p = 0.0102] and severely reduced LV function [HR 5.463 (2.098-14.26); p = 0.0005]. CONCLUSION: An r-ADP-agg ≥ 50% is associated with an increased mortality in patients after coronary stent implantation. Furthermore, r-ADP-agg might represent a better tool to predict clinical outcome than the conventional ADP induced platelet aggregation alone.

Serotonin antagonism improves platelet inhibition in clopidogrel low-responders after coronary stent placement: an in vitro pilot study.

Increased residual platelet reactivity remains a burden for coronary artery disease (CAD) patients who received a coronary stent and do not respond sufficiently to treatment with acetylsalicylic acid and clopidogrel. We hypothesized that serotonin antagonism reduces high on-treatment platelet reactivity. Whole blood impedance aggregometry was performed with arachidonic acid (AA, 0.5 mM) and adenosine diphosphate (ADP, 6.5 µM) in addition to different concentrations of serotonin (1-100 µM) in whole blood from 42 CAD patients after coronary stent placement and 10 healthy subjects. Serotonin increased aggregation dose-dependently in CAD patients who responded to clopidogrel treatment: After activation with ADP, aggregation increased from 33.7 ± 1.3% to 40.9 ± 2.0% in the presence of 50 µM serotonin (p<0.05) and to 48.2 ± 2.0% with 100 µM serotonin (p<0.001). The platelet serotonin receptor antagonist ketanserin decreased ADP-induced aggregation significantly in clopidogrel low-responders (from 59.9 ± 3.1% to 37.4 ± 3.5, p<0.01), but not in clopidogrel responders. These results were confirmed with light transmission aggregometry in platelet-rich plasma in a subset of patients. Serotonin hence increased residual platelet reactivity in patients who respond to clopidogrel after coronary stent placement. In clopidogrel low-responders, serotonin receptor antagonism improved platelet inhibition, almost reaching responder levels. This may justify further investigation of triple antiplatelet therapy with anti-serotonergic agents.