RESUMEN
Multiplex protein imaging technologies enable deep phenotyping and provide rich spatial information about biological samples. Existing methods have shown great success but also harbored trade-offs between various pros and cons, underscoring the persisting necessity to expand the imaging toolkits. Here we present PACIFIC: photoactive immunofluorescence with iterative cleavage, a new modality of multiplex protein imaging methods. PACIFIC achieves iterative multiplexing by implementing photocleavable fluorophores for antibody labeling with one-step spin-column purification. PACIFIC requires no specialized instrument, no DNA encoding, or chemical treatments. We demonstrate that PACIFIC can resolve cellular heterogeneity in both formalin-fixed paraffin-embedded (FFPE) samples and fixed cells. To further highlight how PACIFIC assists discovery, we integrate PACIFIC with live-cell tracking and identify phosphor-p70S6K as a critical driver that governs U87 cell mobility. Considering the cost, flexibility, and compatibility, we foresee that PACIFIC can confer deep phenotyping capabilities to anyone with access to traditional immunofluorescence platforms.
RESUMEN
Amylose-lipid complex (ALC) was prepared with corn starch and stearic acid and used as a shortening replacement in white pan bread preparation. ALCs were prepared using various concentrations of stearic acid to corn starch (1%, 3%, 5%, and 7%) under different temperatures (55, 65, and 75 °C) and for different durations of time (30, 60, and 120 min); then, their complexing properties were assessed using iodine reagent and X-ray diffraction. The complexing reaction at 75 °C for 60 min showed the highest complexing index of the tested conditions; the in vitro digestibility of ALC was lower than that of corn starch. White pan bread was prepared with ALCs and their characteristics, including appearance, loaf volume, and starch retrogradation during storage at room temperature for four days, were compared with those of control bread. With increasing ALC replacement concentrations, loaf volume and shape were significantly affected; however, starch retrogradation was significantly retarded and energy value decreased by ALC replacement. Overall, 50% replacement of shortening by ALC appeared to be a reasonable level for retaining the basic characteristics of the bread while retarding the staling process. These results indicate that ALCs may be potentially useful in the bakery industry for preparing low calorie and low-fat products.