• Molecules and Cells
• /
• v.39 no.6
• /
• pp.439-446
• /
• 2016

Clearing and labeling techniques for large-scale biological tissues enable simultaneous extraction of molecular and structural information with minimal disassembly of the sample, facilitating the integration of molecular, cellular and systems biology across different scales. Recent years have witnessed an explosive increase in the number of such methods and their applications, reflecting heightened interest in organ-wide clearing and labeling across many fields of biology and medicine. In this review, we provide an overview and comparison of existing clearing and labeling techniques and discuss challenges and opportunities in the investigations of large-scale biological systems.

• Journal of the Korean Magnetic Resonance Society
• /
• v.18 no.2
• /
• pp.47-51
• /
• 2014

Larger proteins (above molecular weight 50 kDa) usually show slow motional tumbling in solution, which facilitates the decay of NMR signal, resulting in poor signal-to-noise. In the past twenty years, researchers have tried to overcome this problem with higher molecular weight by improvement of hardware (higher magnetic field and cryoprobe), optimization of pulse sequences for lager molecules, and development of isotope-labeling techniques. Actually, GroEL/ES complex (${\approx}$ 900 kDa) was successfully studied using combination of above techniques. Among the techniques used in large molecular studies, the impact of isotope-labeling for large molecules study is summarized and discussed here.

• Molecules and Cells
• /
• v.26 no.6
• /
• pp.530-535
• /
• 2008

Optical signals based on Raman scattering, coherent anti-Stokes Raman scattering (CARS), and harmonic generation can be used to image biological molecules in living cells without labeling. Both Raman scattering and CARS signals can be used to detect frequencies of molecular vibrations and to obtain the molecular distributions in samples. Second-harmonic optical signals can also be generated in structured arrays of noncentrosymmetric molecules and can be used to detect structured aggregates of proteins, such as, collagen, myosin and tubulin. Since labeling techniques using chemical and biological reactions may cause undesirable changes in the sample, label-free molecular imaging techniques are essential for observation of living samples.

• Journal of Radiopharmaceuticals and Molecular Probes
• /
• v.4 no.1
• /
• pp.36-42
• /
• 2018

Molecular imaging refers to detect the biochemical process in living organisms at the cellular and molecular levels and to quantify them. Due to several advantages of nanomaterials, various molecular images using nanomaterials are being tried. Attempts have been made to combine nanoparticles, known as micro- or nanosized nanomaterials, with radioactive isotopes for molecular imaging probe. The radiolabeled nanoparticles will expend the molecular imaging due to nanoparticle's size-dependent nature. In particular, iron oxide nanoparticles can be used for magnetic resonance imaging, can be adjusted in size, easily functionalized, and biocompatible, making it a very good platform for molecular imaging. In addition, iron oxide nanoparticles may be the best example for a new approach to molecular imaging techniques. In this paper, we introduce various methods for preparation of iron oxide nanoparticle combined with radioisotope starting from various synthesis methods of iron oxide nanoparticles to utilize iron oxide nanoparticles as a platform for molecular imaging through radioactive labeling.

• BMB Reports
• /
• v.28 no.5
• /
• pp.433-436
• /
• 1995

In order to assign NMR signals, conditions for the specific labeling of cytochrome $c_3$ of D. vulgaris Miyazaki F through the culture in a minimal medium were established. Phenylalanine residue was specifically deuterated at more than 85% efficiency. Cytochrome $c_3$ has two phenylalanine residues. The signals of one phenylalane were missing and this was tentatively assigned to Phe20.

• BMB Reports
• /
• v.30 no.3
• /
• pp.211-215
• /
• 1997

A photoactive analog of ATP, 3'(2')-O-(p-azidobenzoyl)-adenosine 5-triphosphate (AB-ATP) was synthesized by chemically coupling N-hydroxysuccinimidyl-4-azidobenzoate (NHS-AB) and ATP. The utility of AB-ATP as an effective active-site-directed photoprobe was demonstrated using catalytic subunit of protein kinase A as a model enzyme. Photoincorporation of AB-ATP was saturated with apparent dissociation constant of $30{\mu}m$ and protected completely by $100{\mu}m$ of ATP. When the enzyme was covalently modified by photolysis in the presence of saturating amounts of photoprobe, about 60% inhibition of enzyme activity was observed. These results demonstrate that AB-ATP has potential application as a probe to characterize ATP-binding proteins including protein kinases.

• Journal of Radiopharmaceuticals and Molecular Probes
• /
• v.1 no.1
• /
• pp.15-22
• /
• 2015

$^{18}F$-labeling reaction of bioactive molecule via click chemistry is widely used to produce $^{18}F$-labeled radiotracer in the field of radiopharmaceutical science and molecular imaging. In particular, bioorthogonal strain-promoted alkyne-azide cycloaddition (SPAAC) reaction has received much attention as an alternative ligation method for radiolabeling bioactive molecules such as peptides, DNA, proteins as well as nanoparticles. Moreover, SPAAC based pretargeting method could provide tumor images successfully on positron emission tomography system using nanoparticle such as mesoporous silica nanoparticles.

• Molecules and Cells
• /
• v.42 no.5
• /
• pp.386-396
• /
• 2019

Labeling of a protein with a specific dye or tag at defined positions is a critical step in tracing the subtle behavior of the protein and assessing its cellular function. Over the last decade, many strategies have been developed to achieve selective labeling of proteins in living cells. In particular, the site-specific unnatural amino acid (UAA) incorporation technique has gained increasing attention since it enables attachment of various organic probes to a specific position of a protein in a more precise way. In this review, we describe how the UAA incorporation technique has expanded our ability to achieve site-specific labeling and visualization of target proteins for functional analyses in live cells.

• Applied Microscopy
• /
• v.46 no.3
• /
• pp.134-139
• /
• 2016

Detailed structural and molecular imaging of intact organs has incurred academic interest because the associated technique is expected to provide innovative information for biological investigation and pathological diagnosis. The conventional methods for volume imaging include reconstruction of images obtained from serially sectioned tissues. This approach requires intense manual work which involves inevitable uncertainty and much time to assemble the whole image of a target organ. Recently, effective tissue clearing techniques including CLARITY and ACT-PRESTO have been reported that enables visualization of molecularly labeled structures within intact organs in three dimensions. The central principle of the methods is transformation of intact tissue into an optically transpicuous and macromolecule permeable state without loss of intrinsic structural integrity. The rapidly evolving protocols enable morphological analysis and molecular labeling of normal and pathological characteristics in large assembled biological systems with single-cell resolution. The deep tissue volume imaging will provide fundamental information about mutual interaction among adjacent structures such as connectivity of neural circuits; meso-connectome and clinically significant structural alterations according to pathologic mechanisms or treatment procedures.

• Journal of Radiopharmaceuticals and Molecular Probes
• /
• v.4 no.2
• /
• pp.45-50
• /
• 2018

Great efforts are currently devoted to the development of new approaches for the labeling of cells using appropriate radionuclides. While fluoride-18 and copper-64 have been extensively studied as short-term and intermediate-term trafficking agents, iodide was studied less intensely. Here, we report a new cell labeling agent labeled with $^{131}I$, $[^{131}I]$oleyl-4-iodobenzoate ($[^{131}I]$OIB) for long-term cell trafficking. A precursor of $[^{131}I]$OIB was obtained in two steps, with the yield of 35%. The radiochemical yield of $[^{131}I]$OIB was over 50%. While $[^{131}I]$OIB could label different cells, L6 cells showed the highest cell-labeling efficiency. The $[^{131}I]$OIB-labeled L6 cells were imprinted into a rat heart, and then monitored noninvasively for 2 weeks by gamma camera imaging. We conclude that $[^{131}I]$OIB is a good candidate molecule for a long-term cell trafficking agent.