Protein-Protein Affinity Determination by Quantitative FRET Quenching.

The molecular dissociation constant, Kd, is a well-established parameter to quantitate the affinity of protein-protein or other molecular interactions. Recently, we reported the theoretical basis and experimental procedure for Kd determination using a quantitative FRET method. Here we report a new development of Kd determination by measuring the reduction in donor fluorescence due to acceptor quenching in FRET. A new method of Kd determination was developed from the quantitative measurement of donor fluorescence quenching. The estimated Kd values of SUMO1-Ubc9 interaction based on this method are in good agreement with those determined by other technologies, including FRET acceptor emission. Thus, the acceptor-quenched approach can be used as a complement to the previously developed acceptor excitation method. The new methodology has more general applications regardless whether the acceptor is an excitable fluorophore or a quencher. Thus, these developments provide a complete methodology for protein or other molecule interaction affinity determinations in solution

In vivo integrated photoacoustic and confocal microscopy of hemoglobin oxygen saturation and oxygen partial pressure

We developed dual-modality microscope integrating photoacoustic microscopy (PAM) and fluorescence confocal microscopy (FCM) to noninvasively image hemoglobin oxygen saturation (sO_2) and oxygen partial pressure (pO_2) in vivo in single blood vessels with high spatial resolution. While PAM measures sO_2 by imaging hemoglobin optical absorption at two wavelengths, FCM quantifies pO_2 using phosphorescence quenching. The variations of sO_2 and pO_2 values in multiple orders of vessel branches under hyperoxic (100% oxygen) and normoxic (21% oxygen) conditions correlate well with the oxygen–hemoglobin dissociation curve. In addition, the total concentration of hemoglobin is imaged by PAM at an isosbestic wavelength

Noninvasive photoacoustic sentinel lymph node mapping using Au nanocages as a lymph node tracer in a rat model

Sentinel lymph node biopsy (SLNB) has been widely performed and become the standard procedure for axillary staging in breast cancer patients. In current SLNB, identification of SLNs is prerequisite, and blue dye and/or radioactive colloids are clinically used for mapping. However, these methods are still intraoperative, and especially radioactive colloids based method is ionizing. As a result, SLNB is generally associated with ill side effects. In this study, we have proposed near-infrared Au nanocages as a new tracer for noninvasive and nonionizing photoacoustic (PA) SLN mapping in a rat model as a step toward clinical applications. Au nanocages have great features: biocompatibility, easy surface modification for biomarker, a tunable surface plasmon resonance (SPR) which allows for peak absorption to be optimized for the laser being used, and capsule-type drug delivery. Au nanocage-enhanced photoacoustic imaging has the potential to be adjunctive to current invasive SLNB for preoperative axillary staging in breast cancer patients

Near-Infrared Gold Nanocages as a New Class of Tracers for Photoacoustic Sentinel Lymph Node Mapping on a Rat Model

This work demonstrated the use of Au nanocages as a new class of lymph node tracers for noninvasive photoacoustic (PA) imaging of a sentinel lymph node (SLN). Current SLN mapping methods based on blue dye and/or nanometer-sized radioactive colloid injection are intraoperative due to the need for visual detection of the blue dye and low spatial resolution of Geiger counters in detecting radioactive colloids. Compared to the current methods, PA mapping based on Au nanocages shows a number of attractive features: noninvasiveness, strong optical absorption in the near-infrared region (for deep penetration), and the accumulation of Au nanocages with a higher concentration than the initial solution for the injection. In an animal model, these features allowed us to identify SLNs containing Au nanocages as deep as 33 mm below the skin surface with good contrast. Most importantly, compared to methylene blue Au nanocages can be easily bioconjugated with antibodies for targeting specific receptors, potentially eliminating the need for invasive axillary staging procedures in addition to providing noninvasive SLN mapping

Near-Infrared Gold Nanocages as a New Class of Tracers for Photoacoustic Sentinel Lymph Node Mapping on a Rat Model

This work demonstrated the use of Au nanocages as a new class of lymph node tracers for noninvasive photoacoustic (PA) imaging of a sentinel lymph node (SLN). Current SLN mapping methods based on blue dye and/or nanometer-sized radioactive colloid injection are intraoperative due to the need for visual detection of the blue dye and low spatial resolution of Geiger counters in detecting radioactive colloids. Compared to the current methods, PA mapping based on Au nanocages shows a number of attractive features: noninvasiveness, strong optical absorption in the near-infrared region (for deep penetration), and the accumulation of Au nanocages with a higher concentration than the initial solution for the injection. In an animal model, these features allowed us to identify SLNs containing Au nanocages as deep as 33 mm below the skin surface with good contrast. Most importantly, compared to methylene blue Au nanocages can be easily bioconjugated with antibodies for targeting specific receptors, potentially eliminating the need for invasive axillary staging procedures in addition to providing noninvasive SLN mapping

Photoacoustic quantification of the optical absorption cross-sections of gold nanostructures

This study demonstrates a method for measuring the optical absorption cross-sections (σ_a) of Au-Ag nanocages and Au nanorods using photoacoustic (PA) sensing. PA signals are directly proportional to the absorption coefficient (μ_a) of the nanostructure. For each type of nanostructure, we first obtained μa from the PA signal by benchmarking against a linear calibration curve (PA signal vs. μ_a) derived from a set of methylene blue solutions with different concentrations. We then calculated σ_a by dividing the μ_a by the corresponding concentration of the Au nanostructure. Additionally, we obtained the extinction cross-section (σ_e, sum of absorption and scattering cross-sections) from the extinction spectrum recorded using a conventional UV-vis-NIR spectrometer. From the measurements of σ_a and σ_e, we were able to easily derive both the absorption and scattering cross-sections for each type of gold nanostructure. This method can potentially provide the optical absorption and scattering properties of gold nanostructures and other types of nanomaterials

Measuring the Optical Absorption Cross Sections of Au−Ag Nanocages and Au Nanorods by Photoacoustic Imaging

This paper presents a method for measuring the optical absorption cross sections (σ_a) of Au−Ag nanocages and Au nanorods. The method is based on photoacoustic (PA) imaging, where the detected signal is directly proportional to the absorption coefficient (μ_a) of the nanostructure. For each type of nanostructure, we first obtained μ_a from the PA signal by benchmarking against a linear calibration curve (PA signal versus μ_a) derived from a set of methylene blue solutions with different concentrations. We then calculated σ_a by dividing the μ_a by the corresponding concentration of the Au nanostructure. Additionally, we obtained the extinction cross section (σ_e, sum of absorption and scattering) from the extinction spectrum recorded using a conventional UV−vis−NIR spectrometer. From the measurements of σ_a and σ_e, we were able to easily derive both the absorption and scattering cross sections for each type of gold nanostructure. The ratios of absorption to extinction obtained from experimental and theoretical approaches agreed well, demonstrating the potential use of this method in determining the optical absorption and scattering properties of gold nanostructures and other types of nanomaterials

Noninvasive photoacoustic sentinel lymph node mapping using Au nanocages as a lymph node tracer in a rat model

Sentinel lymph node biopsy (SLNB) has been widely performed and become the standard procedure for axillary staging in breast cancer patients. In current SLNB, identification of SLNs is prerequisite, and blue dye and/or radioactive colloids are clinically used for mapping. However, these methods are still intraoperative, and especially radioactive colloids based method is ionizing. As a result, SLNB is generally associated with ill side effects. In this study, we have proposed near-infrared Au nanocages as a new tracer for noninvasive and nonionizing photoacoustic (PA) SLN mapping in a rat model as a step toward clinical applications. Au nanocages have great features: biocompatibility, easy surface modification for biomarker, a tunable surface plasmon resonance (SPR) which allows for peak absorption to be optimized for the laser being used, and capsule-type drug delivery. Au nanocage-enhanced photoacoustic imaging has the potential to be adjunctive to current invasive SLNB for preoperative axillary staging in breast cancer patients

Water management affects arsenic and cadmium accumulation in different rice cultivars

Paddy rice (Oryza sativa L.) is a staple food and one of the major sources of dietary arsenic (As) and cadmium (Cd) in Asia. A field experiment was conducted to investigate the effects of four water management regimes (aerobic, intermittent irrigation, conventional irrigation and flooding) on As and Cd accumulation in seven major rice cultivars grown in Zhejiang province, east China. With increasing irrigation from aerobic to flooded conditions, the soil HCl-extractable As concentrations increased significantly and the HCl-extractable Cd concentrations decreased significantly. These trends were consistent with the As and Cd concentrations in the straw, husk and brown rice. Water management both before and after the full tillering stage affected As and Cd accumulation in the grains. The intermittent and conventional treatments produced higher grain yields than the aerobic and flooded treatments. Cd concentrations in brown rice varied 13.1-40.8 times and As varied 1.75-8.80 times among the four water management regimes. Cd and As accumulation in brown rice varied among the rice cultivars, with Guodao 6 (GD6) was a low Cd but high-As-accumulating cultivar while Indonesia (IR) and Yongyou 9 (YY9) were low As but high-Cd-accumulating cultivars. Brown rice Cd and As concentrations in the 7 cultivars were significantly negatively correlated. The results indicate that As and Cd accumulated in rice grains with opposite trends that were influenced by both water management and rice cultivar. Production of 'safe' rice with respect to As and Cd might be possible by balancing water management and rice cultivar according to the severity of soil pollution.Paddy rice (Oryza sativa L.) is a staple food and one of the major sources of dietary arsenic (As) and cadmium (Cd) in Asia. A field experiment was conducted to investigate the effects of four water management regimes (aerobic, intermittent irrigation, conventional irrigation and flooding) on As and Cd accumulation in seven major rice cultivars grown in Zhejiang province, east China. With increasing irrigation from aerobic to flooded conditions, the soil HCl-extractable As concentrations increased significantly and the HCl-extractable Cd concentrations decreased significantly. These trends were consistent with the As and Cd concentrations in the straw, husk and brown rice. Water management both before and after the full tillering stage affected As and Cd accumulation in the grains. The intermittent and conventional treatments produced higher grain yields than the aerobic and flooded treatments. Cd concentrations in brown rice varied 13.1-40.8 times and As varied 1.75-8.80 times among the four water management regimes. Cd and As accumulation in brown rice varied among the rice cultivars, with Guodao 6 (GD6) was a low Cd but high-As-accumulating cultivar while Indonesia (IR) and Yongyou 9 (YY9) were low As but high-Cd-accumulating cultivars. Brown rice Cd and As concentrations in the 7 cultivars were significantly negatively correlated. The results indicate that As and Cd accumulated in rice grains with opposite trends that were influenced by both water management and rice cultivar. Production of 'safe' rice with respect to As and Cd might be possible by balancing water management and rice cultivar according to the severity of soil pollution

In vivo integrated photoacoustic and confocal microscopy of hemoglobin oxygen saturation and oxygen partial pressure

We developed dual-modality microscope integrating photoacoustic microscopy (PAM) and fluorescence confocal microscopy (FCM) to noninvasively image hemoglobin oxygen saturation (sO_2) and oxygen partial pressure (pO_2) in vivo in single blood vessels with high spatial resolution. While PAM measures sO_2 by imaging hemoglobin optical absorption at two wavelengths, FCM quantifies pO_2 using phosphorescence quenching. The variations of sO_2 and pO_2 values in multiple orders of vessel branches under hyperoxic (100% oxygen) and normoxic (21% oxygen) conditions correlate well with the oxygen–hemoglobin dissociation curve. In addition, the total concentration of hemoglobin is imaged by PAM at an isosbestic wavelength