Advanced fluorescent chemical probes for detecting metabolites
Abstract
Goryo Chemical Inc. creates innovative fluorescent probes through relationships with foremost scientists in Tokyo ( Prof. Nagano and Prof. Urano) and Hokkaido University ( Nobel prize winner Prof. Suzuki). The chemical probes are nontoxic, highly sensitive and remarkably stable for selective staining of target cells. Also, it can be applied to fluorescent imaging and selection of live cell and tissue without fixation.
Researchers can distinguish target cells from other cells using ProteoGREEN™-gGlu(Cancer cell), KP-1(hiPS/ES cell), MAR (Hypoxia), GlycoYELLOW™ -β Gal (Senescent cell)
ProteoGREEN™-gGlu is cancer-selective probe with intramolecular spirocyclic caging for complete quenching. Activation occurs by rapid one-step cleavage of glutamate with g-glutamyltranspeptidase(GGT), which is not expressed in normal tissue. In vitro activity was evident in 11 human ovarian cancer cell lines. Moreover, it can detect cancer for fluorescent image guided surgery at pre-clinical and clinical trials in JAPAN.
Kyoto probe 1 (KP-1) selectively labels embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs). Selective elimination of human pluripotent stem cells can be achieved by KP-1 with appropriate ABC-transporter (ABCB1 and ABCG2).
MAR features a higher sensitivity and can detect mild hypoxia, as compared to pimonidazole. Reduction of MAR by reductase under hypoxia resulted in the generation of highly fluorescent rhodamine derivatives, 2Me RG.
GlycoYELLOW™-βGal is a specific fluorescent probe for detection of β-galactosidase. It can detect not only fluorescent imaging and selection of live cell and tissue transfected with lacZ but also SA-beta-gal (senescence-associated activity in living cells.
Based on independent technologies, Goryo chemical Inc. has developed advanced small-molecule probes for live cell imaging and helps researchers who want examine cellular function in physiological condition to get more reliable and relevant data.
High-Throughput MALDI Imaging of Tissue: Taking Proteomics into the 2nd and 3rd Dimensions
Abstract
MALDI Imaging mass spectrometry (IMS) is a unique analytical tool that allows simultaneous label-free visualization of hundreds of endogenous compounds expressed in tissue. In combination with histology MALDI-TOF IMS can reveal correlation of many lipids/peptides/proteins to pathological features, creating a multi-dimensional scale for molecular histology that promises to aid in our understanding of disease diagnosis and treatment. Further, MALDI imaging following enzymatic reactions on tissue provide additional depth to the molecular information that can be extracted. TOF mass analyzers provide the widest analyte versatility for MALDI imaging but existing system designs are incapable of meeting the demands for acquiring imaging data sets at a time consistent with clinical relevance. Here we describe a newly designed MALDI-TOF imaging system, rapifleX MALDI Tissuetyper, which offers a significant speed advantage over existing systems.
The new rapifleX MALDI-TOF platform is capable of acquiring up to 50 spectra per second in imaging mode. A novel smartbeam 3D laser produces a beam focused to 5 µm which probes the complete pixel area while the target is in constant motion.
Using rapifleX, images of several hundred thousand pixels were acquired in 1-2 hours, a time scale relevant for clinical applications. Areas of the sample that correspond to pixels are square in shape and can be varied in size from 10-200 µm. The unique combination of moving laser and sample ensures maximum analytical sampling of the complete pixel area and that image pixels contain molecular signals unique to that area of the sample and do not contain signals from adjacent pixels as happens with oversampling techniques. 10 µm resolution MALDI images of phospholipids in excess of 800k pixels have been acquired in 3 hours. 20 µm protein images from large biopsies, >1 cm2, can be acquired in 2-4 hours. Megapixel ion images have been acquired in only a few hours. Images from more than one hundred serial sections from rat kidney were acquired in <48 hrs, permitting 3-dimensional MALDI imaging to be carried out in a practical time-frame.
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