3D spin-lock imaging of human gliomas

We investigated whether the simultaneous use of paramagnetic contrast medium and 3D on-resonance spin lock (SL) imaging could improve the contrast of enhancing brain tumors at 0.1 T. A phantom containing serial concentrations of gadopentetate dimeglumine (Gd-DTPA) in cross-linked bovine serum albumin (BSA) was imaged. Eleven patients with histologically verified glioma were also studied. T₁-weighted 3D gradient echo images with and without SL pulse were acquired before and after a Gd-DTPA injection. SL effect, contrast, and contrast-to-noise ratio (CNR) were calculated for each patient. In the glioma patients, the SL effect was significantly smaller in the tumor than in the white and gray matter both before (p = 0.001, p = 0.025, respectively), and after contrast medium injection (p < 0.001, p < 0.001, respectively). On post-contrast images, SL imaging significantly improved tumor contrast (p = 0.001) whereas tumor CNR decreased slightly (p = 0.024). The combined use of SL imaging and paramagnetic Gd-DTPA contrast agent offers a modality for improving tumor contrast in magnetic resonance imaging (MRI) of enhancing brain tumors. 3D gradient echo SL imaging has also shown potential to increase tissue characterization properties of MR imaging of human gliomas.     

Feasibility of 3D harmonic contrast imaging

Improved endocardial border delineation with the application of contrast agents should allow for less complex and faster tracing algorithms for left ventricular volume analysis. We developed a fast rotating phased array transducer for 3D imaging of the heart with harmonic capabilities making it suitable for contrast imaging. In this study the feasibility of 3D harmonic contrast imaging is evaluated in vitro. A commercially available tissue mimicking flow phantom was used in combination with Sonovue. Backscatter power spectra from a tissue and contrast region of interest were calculated from recorded radio frequency data. The spectra and the extracted contrast to tissue ratio from these spectra were used to optimize the excitation frequency, the pulse length and the receive filter settings of the transducer. Frequencies ranging from 1.66 to 2.35 MHz and pulse lengths of 1.5, 2 and 2.5 cycles were explored. An increase of more than 15 dB in the contrast to tissue ratio was found around the second harmonic compared with the fundamental level at an optimal excitation frequency of 1.74 MHz and a pulse length of 2.5 cycles. Using the optimal settings for 3D harmonic contrast recordings volume measurements of a left ventricular shaped agar phantom were performed. Without contrast the extracted volume data resulted in a volume error of 1.5%, with contrast an accuracy of 3.8% was achieved. The results show the feasibility of accurate volume measurements from 3D harmonic contrast images. Further investigations will include the clinical evaluation of the presented technique for improved assessment of the heart.     

Measurement of UV absorption of single living cell for cell manipulation using NIR femtosecond laser

Optical UV absorption of single human living cells ranging from 200 nm to 360 nm was measured in situ for the study of cell manipulation using the near-infrared (NIR) femtosecond laser. Human breast living cells of MCF-10A, MCF-7, and MDA-MB-231 were used in this experiment. The selective photo-disruptions of single living cell and its sub-organelle (nucleus) were also demonstrated using the tightly focused 790 nm wavelength femtosecond laser with pulse duration of 110 fs. It was found that each living cell has its own absorption spectrum in UV wavelength ranges. It was also inferred that intrinsic absorption spectrum is attributed to the amount of DNA and protein of living cell. For the study of photo-disruption of single cell using the multi-photon absorption excited by the NIR femtosecond laser pulse, the origin UV absorption spectrum of targeted living cell is important and fundamental information to understand nonlinear interaction between NIR ultrashort, high-intensity laser light and transparent living cell.     

S波段微波热致超声成像系统研究

微波热致超声成像技术通过向物体发射微波脉冲, 导致物体吸收电磁波温度迅速升高, 产生瞬时压力波, 从而激发产生超声波信号, 通过传感器对产生的超声波信号进行采集并成像, 最终还原了反映物体吸收电磁波能量特性的图像, 由于此方法兼具了微波成像的高对比性和超声成像的高分辨率特点, 理论上验证了热声成像技术对早期乳腺肿瘤检测的可行性. 本实验兼顾系统成像深度和分辨率, 采用S波段的微波脉冲信号源对物体进行辐射, 利用圆形扫描方式对待测物体进行检测, 同时为了更好的验证成像性能, 本实验同时使用了肿瘤仿体及实际生物组织进行成像实验. 通过实验分析, 验证了该系统对肿瘤仿体和生物组织检测的有效性, 以及系统的高分辨率和高对比度特性, 为早期乳房肿瘤检测提供了进一步的理论支撑.     

Endoscopic, rapid near-infrared optical tomography

This is believed to be the first demonstration of near-infrared (NIR) optical tomography employed at the endoscope scale and at a rapid sampling speed that allows translation to in vivo use. A spread-spectral-encoding technique based on a broadband light source and linear-to-circular fiber bundling was used to provide endoscopic probing of many source-detector fibers for tomography as well as parallel sampling of all source-detector pairs for rapid imaging. Endoscopic NIR tomography at an 8 Hz frame rate was achieved in phantoms and tissue specimens with a 12 mm probe housing eight sources and eight detectors. This novel approach provides the key feasibility studies to allow this blood-based contrast imaging technology to be attempted in detection of cancer in internal organs via endoscopic interrogation.     

聚吡咯修饰的金纳米棒对肿瘤的光学相干层析成像的影响

聚吡咯(PPy)制备简单、生物相容性好,且在近红外(NIR)光谱范围内有很强的吸收,可作为一种良好的光热治疗试剂;同时,其NIR光吸收性质也可用于增强光学相干层析成像(OCT)的对比效果。因此,采用PPy对传统的OCT对比试剂——金纳米棒(GNR)进行表面修饰,有望获得对比效果更好且生物毒性较小的新型OCT对比试剂。选用吡咯为起始原料,在GNR表面进行一步简单的氧化聚合反应即可制备得到PPy修饰的金纳米棒(GNR-PPy)。利用紫外-可见吸收光谱,拉曼光谱和透射电子显微镜对制备的样品进行了分析和表征。构建小鼠荷瘤模型,以研究GNR-PPy对肿瘤OCT图像对比度的增强效果。采用中心波长为840 nm的OCT系统对注射了纳米粒子的肿瘤区域进行OCT成像。结果表明,肿瘤组织注射了GNR-PPy后,OCT信号衰减非常明显;与注射了GNR的OCT图像相比,840 nm光在GNR-PPy的OCT图像中的穿透深度明显更低。从OCT图像中抽提出一维的衰减曲线对OCT图像进行定量分析,发现注射有GNR-PPy肿瘤组织的OCT信号衰减系数明显高于注射了GNR的组织。表明,相对于GNR,GNR-PPy具有更好的OCT信号对比效果,这在增强肿瘤成像效果方面具有潜在应用价值。     

Phase contrast MRI with improved temporal resolution by view sharing: k-space related velocity mapping properties.

Phase contrast techniques in combination with k-space segmented CINE imaging are widely used for the quantitative assessment of blood flow or tissue motion. The temporal resolution of the corresponding pulse sequences plays an important role concerning the potential of the method to fully detect time resolved flow or motion patterns. A further improvement of temporal or spatial resolution in phase contrast CINE MRI can be achieved by the application of view sharing. Based on simulations with point-spread-functions resulting from different cyclic flow or motion patterns an analysis of view sharing techniques in combination with phase contrast MRI is presented. Velocity mapping properties and the role of different k-space regions concerning the resulting values in the phase images and thus encoded velocities were investigated. It could be shown that the velocity induced phase shifts in phase contrast techniques are mainly encoded in the central sections of k-space which makes view sharing also suitable for velocity mapping. As a result the use of appropriate sampling and data acquisition schemes permits the assessment of flow or motion patterns with significantly improved temporal resolution without loss of functional information. In addition phantom measurements with an oscillation phantom were performed in order to validate the simulation results and to demonstrate the potential of view sharing techniques to accelerate phase contrast imaging and improve the detection of the underlying flow or motion dynamics.     

Magnetization Transfer Imaging of Rat Brain under Non-steady-state Conditions. Contrast Prediction Using a Binary Spin–Bath Model and a Super-Lorentzian Lineshape

Magnetization transfer contrast imaging using turbo spin echo and continuous wave off-resonance irradiation was carried out on rat brainin vivoat 4.7 T. By systematically varying the off-resonance irradiation power and the offset-frequency, the signal intensities obtained under steady-state for both transverse and longitudinal magnetization were successfully analyzed with a simple binary spin–bath model taking into account a free water compartment and a pool of protons with restricted motions bearing a super-Lorentzian lineshape. Due to important RF power deposition, such experimental conditions are not practical for routine imaging on humans. An extension of the model was derived to describe the system for shorter off-resonance pulse duration, i.e., when the longitudinal magnetization of the free protons has not reached a steady-state. Data sets obtained for three regions of interest, namely thecorpus callosum,the basal ganglia, and the temporal lobe, were correctly interpreted for off-resonance pulse durations varying from 0.3 to 3 s. The parameter sets obtained from the calculations made it possible to predict the contrast between the different regions as a function of the pulse power, the offset frequency, and pulse duration. Such an approach could be extended to contrast prediction for human brain at 1.5 T.     

Photoacoustic Tomography of Biological Sample Using Phase-controlled Focus Algorithm

Photoacoustic (PA) imaging is a new imaging modality, which converts pressure signals received by a scanning detector to a local distribution of electromagnetic absorption density. In this paper an experiment result of a photoacoustic tomography to depths of ～7 mm for a real tissue is presented, using a 532-nm pulse YAG laser. The time-resolved stress detection technique was used for PA signal detection with a high temporal resolution. A phase-controlled focus algorithm was used for image reconstruction. Images of different depth profiles in tissue were obtained. The depth resolution was 30 μm and could be up to 10 μm using a wide-band tranducer.     

Photoacoustic Tomography of Biological Sample Using Phase-controlled Focus Algorithm

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In situ observation of photo-bleaching in human single living cell excited by a NIR femtosecond laser

The photo-bleaching of single living cells excited by femtosecond laser irradiation was observed in situ to study the nonlinear interaction between ultrafast laser pulses and living human breast MDA-MB-231 cells. We conducted a systematic study of the energy dependence of plasma-mediated photo-disruption of fluorescently labeled subcellular structures in the nucleus of living cells using near-infrared (NIR) femtosecond laser pulses through a numerical aperture objective lens (0.75 NA). The behavior of photo-bleached living cells with fluorescently labeled nuclei was observed for 18 h after femtosecond laser irradiation under a fluorescence microscope. The photo-bleaching of single living cells without cell disruption occurred at between 470 and 630 nJ. To study the photo-disruption of subcellular organelles in single living cells using the nonlinear absorption excited by a NIR femtosecond laser pulse, the process of photo-bleaching without photo-disruption provides key information for clarifying the nonlinear interaction between NIR ultrashort, high-intensity laser light and transparent fluorescently labeled living cells.     

Fast 3D echo planar SSFP-based 1H spectroscopic imaging: demonstration on the rat brain in vivo

A fast proton spectroscopic imaging pulse sequence based on the condition of steady-state free precession is presented. High 3D spatial and temporal resolution is achieved using simultaneous detection of both one spatial and one spectral dimension, with a time-dependent gradient cycle known from echo planar imaging. Additionally, in order to increase the spectral width of the measurement, an interleaved acquisition scheme is shown either for systems with limited gradient switching capabilities or applications with a wide chemical shift range. The pulse sequence is implemented on a standard 4.7-T nuclear magnetic resonance animal imaging system. Measurements with a total measurement time of less than 2.5 min and a nominal voxel size of 6.75 microl using a total of 64 x 32 x 16 voxels are performed on phantoms and healthy rat brain in vivo allowing the rapid detection of signals from both uncoupled and J-coupled spin systems with high signal-to-noise ratio.     

Spectral-domain optical coherence phase and multiphoton microscopy

We describe simultaneous quantitative phase contrast and multiphoton fluorescence imaging by combined spectral-domain optical coherence phase and multiphoton microscopy. The instrument employs two light sources for efficient optical coherence microscopic and multiphoton imaging and can generate structural and functional images of transparent specimens in the epidirection. Phase contrast imaging exhibits spatial and temporal phase stability in the subnanometer range. We also demonstrate the visualization of actin filaments in a fixed cell specimen, which is confirmed by simultaneous multiphoton fluorescence imaging.     

NIR‐Triggered Release of Nitric Oxide with Upconversion Nanoparticles Inhibits Platelet Aggregation in Blood Samples

There is a great challenge to overcome the limitation of tissue penetration depth, while maximizing the benefit of light‐triggered biochemical cascades in a well‐defined mode simultaneously. Here, a new method of near‐infrared (NIR) light‐triggered release of nitric oxide (NO) by developing upconversion nanoparticles (UCNPs)‐based conjugate chemistry is reported. As the key nanotransducer in the design, core–shell‐structured UCNPs are encapsulated with a layer of SiO₂ and then covalently linked with a potent NO‐releasing donor (S‐nitroso‐N‐acetyl‐dl ‐penicillamine, SNAP). It is featured with highly localized breakage of chemical bonds of SNAP molecules by NIR–UV upconversion, enabling simultaneous NO release in a light dosage‐dependent manner. The biological effects of NO releasing are demonstrated by cellular imaging and inhibition of platelet aggregation from blood samples. This work provides a flexible and robust platform to generate cell‐signaling gas molecules trigged by NIR laser with deep tissue penetration.     

Optimizing tissue contrast in magnetic resonance imaging

Magnetic resonance imaging demands that tissue contrast and signal-to-noise advantages be sought in each component of the imaging system. One component of magnetic resonance imaging in which contrast and signal-to-noise ratios are easily manipulated is in the choice of pulse sequences and interpulse delay times. This article provides a general method for determining the best choices of interpulse delay times in pulse sequences and applies that method to saturation recovery, inversion recovery, and spin-echo sequences. Saturation recovery and inversion recovery sequences with rephasing pulses, and tissues with unequal hydrogen densities are considered. Optimization of pulse sequences is carried out for the two distinct cases of (a) a fixed number of sequence repetitions and (b) a fixed total imaging time. Analytic expressions are derived or approximate expressions are provided for the interpulse delay times that optimize contrast-to-noise ratios in each pulse sequence. The acceptable range of interpulse delay times to obtain reasonable contrast using each pulse sequence is discussed.     

乳腺癌的临床光谱检测

基于激光与生物组织的相互作用原理检测乳腺癌是目前国际研究热点。实验室自行开发研制的近红外乳腺检测仪, 采用780 nm的低频调制激光透照乳房,并在计算机控制下做二维扫描,对侧采用光电倍增管接收透射光信号,将其转化为电信号,实现了对乳腺癌疾病的检测。系统对光强信号采样后进行数据处理和成像,得到近红外光乳腺透照图像。该系统在中国医学科学院肿瘤医院进行了临床试验。共采集了50名患者100多个数据。其中恶性肿瘤患者34例,良性13例,其他3例。近红外诊断结果与患者同期的钼靶检测结果、超声检测结果和病理结果做了对照,正确率达到72.5%,介于超声(77.50%)和钼靶(71.88%)之间。说明近红外方法和所使用的样机对判断乳腺疾病有可行性,为近红外图像方法提供了新的研究思路。     

In vivo near-infrared spectral detection of pressure-induced changes in breast tissue

A diffuse near-infrared tomography system was used to measure dynamic changes in the absolute optical properties of the human breast that were induced through pressure applied to the tissue surface. Results from five subjects show that absorption and scattering coefficients changed measurably when pressure was increased and that these relative changes correlated with the subjects' body-mass index, indicating that the effect depends on tissue composition. Fitting the absolute absorption and scattering coefficients at six wavelengths to the molar absorption spectra of the three predominant chromophores revealed that both the average total hemoglobin and oxygen saturation increased by 10%, while water concentration decreased by more than 12%. These changes indicate that the pressure-induced variation is likely due to water displacement and vascular volume increase in the region being imaged, for mild application of pressure to the breast. These results suggest that the pressure applied during optical measurements of tissue may alter the tissue physiology, and care should be taken to factor this effect into the design of optical medical instrumentation. In addition, the technique provides a unique approach to measuring tissue elastic changes in vivo in the female breast and may offer a new method for dynamic contrast imaging based on elasto-optical measurements.     

A UV–Visible–NIR fluorescence lifetime imaging microscope for laser-based biological sensing with picosecond resolution

This article describes the design and characterization of a wide-field, time-domain fluorescence lifetime imaging microscopy (FLIM) system developed for picosecond time-resolved biological imaging. The system consists of a nitrogen-pumped dye laser for UV–visible–NIR excitation (337.1–960 nm), an epi-illuminated microscope with UV compatible optics, and a time-gated intensified CCD camera with an adjustable gate width (200 ps-10^-3 s) for temporally resolved, single-photon detection of fluorescence decays with 9.6-bit intensity resolution and 1.4-μm spatial resolution. Intensity measurements used for fluorescence decay calculations are reproducible to within 2%, achieved by synchronizing the ICCD gate delay to the excitation laser pulse via a constant fraction optical discriminator and picosecond delay card. A self-consistent FLIM system response model is presented, allowing for fluorescence lifetimes (0.6 ns) significantly smaller than the FLIM system response (1.14 ns) to be determined to 3% of independently determined values. The FLIM system was able to discriminate fluorescence lifetime differences of at least 50 ps. The spectral tunability and large temporal dynamic range of the system are demonstrated by imaging in living human cells: UV-excited endogenous fluorescence from metabolic cofactors (lifetime ∼1.4 ns); and 460-nm excited fluorescence from an exogenous oxygen-quenched ruthenium dye (lifetime ∼400 ns). Received: 23 February 2003 / Published online: 22 May 2003 RID="*" ID="*"Corresponding author. Fax: +1-734/9361-905, E-mail: mycek@umich.edu     

Early Detection of Breast Cancer: Synthesis and Characterization of Novel Target Specific NIR-Fluorescent Estrogen Conjugate for Molecular Optical Imaging

Estrogen induced proliferation of existing mutant cells is widely understood to be the major risk determining factor in the development of breast cancer. Hence determination of the Estrogen Receptor[ER] status is of paramount importance. We have carried out the synthesis and characterization of a novel NIR fluorescent dye conjugate aimed at measuring ER+ve status in-vivo. The conjugate was synthesized by ester formation between 17-β estradiol and a cyanine dye namely: bis-1, 1-(4-sulfobutyl) indotricarbocyanine-5-carboxylic acid, sodium salt. The replacement of the sodium ion in the ester by a larger glucosammonium ion was found to enhance the hydrophilicity and reduce the toxic effect on cell lines. The excitation and emission peaks for the dye were recorded as 750 and 788 nm respectively; ideal for non-invasive optical imaging owing to minimal tissue attenuation and auto-fluorescence at these wavelengths. The dye (NIRDC1) has a significant drop in plasma-protein binding therefore leading to marked improvement in pharmacokinetic profile such as dye evacuation in comparison to ICG. In addition the dye showed enhanced fluorescence quantum yield, molar extinction coefficient and linearity in fluorescence relative to ICG. This dye can be potentially used as a target specific exogenous contrast agent in molecular optical imaging for early detection of breast cancer.     

Gold nanoshell bioconjugates for molecular imaging in living cells

Advances in scattering-based optical imaging technologies offer a new approach to noninvasive point-of-care detection, diagnosis, and monitoring of cancer. Emerging photonics technologies provide a cost-effective means to image tissue in vivo with high resolution in real time. Advancing the clinical potential of these imaging strategies requires the development of optical contrast agents targeted to specific molecular signatures of disease. We describe the use of a novel class of contrast agents based on nanoshell bioconjugates for molecular imaging in living cells. Nanoshells offer significant advantages over conventional imaging probes including continuous and broad wavelength tunability, far greater scattering and absorption coefficients, increased chemical stability, and improved biocompatibility. We show that nanoshell bioconjugates can be used to effectively target and image human epidermal growth factor receptor 2 (HER2), a clinically relevant biomarker, in live human breast carcinoma cells.