Molecular imaging assays

The aim of our work was to develop a fast, simple and selective CL imaging assay coupled with MIP for the chiral recognition of fluorescence labeled phenylalanine. The precipitation polymerization method was used for preparing MIP microspheres with uniform shape. Figure 2 shows the dansyl-L-phenylalanine molecular imprinting process. The average diameter of microspheres was about 0.7 pm. 

Wang L, Zhang Z. Chemiluminescence imaging assay dipyridamole based on molecular imprinted polymer as recognition material. Sens Actuators B, on line - DOI 10.1016/j.snb.2001.01.051. 

Step 2 SPADS interaction and SPADS imaging assays. FTIR microspectroscopy is used to study intermolecular interactions. Atomic force microscopy (AFM) is used to examine the molecular homogeneity (uniformity) of ASD and the effect of physical stress on uniformity. 

Fig. 3. Comparison between IP-One kit versus calcium mobilization assay (384-well format). Human embryonic kidney (HEK) 293 cells expressing a chemokine receptor were evaluated on HTRF IP-One kit (CisBio, Bedford, MA) and fluroescent imaging plate reader (FLIPR) with Calcium 3 kit (Molecular Devices, Mountain View, CA).

Figure 4. DDC (A), serotonin (B), and tyrosine hydroxylase (C) immunore-activity in the posterior region of a wild-type Drosophila ventral ganglion. Tyrosine hydroxylase (TH) encodes the rate-limiting step in dopamine biosynthesis and is a marker for dopamine cells. B and C are the same CNS assayed for both serotonin and TH. M, medial dopamine neurons VL, ventrolateral serotonin neurons DL, dorsolateral dopamine neurons. Short unmarked arrows in C show vacuolated cells that do not contain DDC immunoreactivity. The immunoreactivity in these cells may represent a nonspecific cross-reactivity of the rat TH antibody. The length bar in A is 50 pM. The images are confocal projections generated on a Molecular Dynamics-2000 confocal laser scanning microscope.

Measuring FRET by fluorescence lifetime imaging microscopy (FRET-FLIM) offers the ability to see beyond the resolution of the optical system ( 10-100 times that of modern far field microscopes [5]). FRET efficiency can be used as a proxy for molecular distance, thereby allowing the easy detection and somewhat more challenging quantification of molecular interactions. Although many types of assay exist, FRET-FLIM is a highly suitable technique that is capable of in situ measurements of molecular interactions and conformation in living and fixed cells. 

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