Optical penetration of surface-enhanced micro-scale spatial offset Raman spectroscopy in turbid gel and biological tissue

The limited penetration of photons in biological tissue restricts the deep-tissue detection and imaging application. The micro-scale spatially offset Raman spectroscopy (micro-SORS) with an optical fiber probe, colleting photons from deeper regions by offsetting the position of laser excitation from the collection optics in a range of hundreds of microns, shows great potential to be integrated with endoscopy for inside-body noninvasive detection by circumventing this restriction, particularly with the combination of surface-enhanced Raman spectroscopy (SERS). However, a detailed tissue penetration study of micro-SORS in combination with SERS is still lacking. Herein, we compared the signal decay of enhanced Raman nanotags through the tissue phantom of agarose gel and the biological tissue of porcine muscle in the near-infrared (NIR) region using a portable Raman spectrometer with a micro-SORS probe (2.1[Formula: see text]mm in diameter) and a conventional hand-held probe (9.7[Formula: see text]mm in diameter). Two kinds of Raman nanotags were prepared from gold nanorods decorated with the nonresonant (4-nitrobenzenethiol) or resonant Raman reporter molecules (IR-780 iodide). The SERS measurements show that the penetration depths of two Raman nanotags are both over 2[Formula: see text]cm in agarose gel and 3[Formula: see text]mm in porcine muscle. The depth could be improved to over 4[Formula: see text]cm in agarose gel and 5[Formula: see text]mm in porcine tissue when using the micro-SORS system. This demonstrates the superiority of optical-fiber micro-SORS system over the conventional Raman detection for the detection of nanotags in deeper layers in the turbid medium and biological tissue, offering the possibility of combining the micro-SORS technique with SERS for noninvasive in vivo endoscopy-integrated clinical application.

Immunochromatographic Detection of Myoglobin as a Specific Biomarker of Porcine Muscle Tissues in Meat Products

An immunochromatographic detection of myoglobin (MG) as a specific marker of porcine muscle tissue has been developed. The method is based on the sandwich lateral flow immunoassay (LFIA) with gold nanoparticles (AuNPs) as a label. The developed test system determines MG with a detection limit of 5 ng mL−1 within 15 min. A specific determination of porcine MG and no cross-reactivity with MG from other tested mammals and bird species was demonstrated. The test system is able to detect pork additives, as low as 0.01% (w/w), in minced beef. A technique of MG extraction from muscle tissue has been proposed which allows for rapid and efficient MG extraction from meat samples (within 20 min). The developed test system can serve as an effective means of controlling the authenticity and quality of meat products.

Isolation and ex vivo cultivation of single myofibers from porcine muscle

The isolation and cultivation of intact, single myofibers presents a superior approach for studying myogenic cells in their native position. The cells’ characteristics remain more similar to muscle tissue than in cell culture. Nevertheless, no routinely used method in higher vertebrates exists. Therefore, we aimed at establishing the isolation and cultivation of single myofibers from porcine muscle. For the first time, we implemented the isolation of intact myofibers from porcine fibularis tertius muscle by enzymatic digestion and their subsequent cultivation under floating conditions. Confocal microscopy showed intact myofibrill structures in isolated myofibers. Myogenic cells were able to proliferate at their parent myofiber as shown by the increase of myonuclear number during culture. Additionally, the described method can be used to investigate myogenic cells migrated from isolated myofibers. These cells expressed myogenic markers and were able to differentiate. In the future, our method can be used for genetic manipulation of cells at myofibers, investigation of growth factors or pharmacological substances, and determination of interactions between myofibers and associated cells. Working with isolated myofibers has the potential to bridge conventional cell culture and animal experiments. Adapting the method to porcine muscle allows for application possibilities in veterinary medicine as well as in biomedical research, which cannot be addressed in rodent model systems.