Use of glycerol as an optical clearing agent for enhancing photonic transference and detection of Salmonella typhimurium through porcine skin

Abstract

The objective of this study was to evaluate glycerol (GLY) and GLY + dimethyl sulfoxide (DMSO) to increase photonic detection of transformed Salmonella typhimurium (S. typh-lux) through porcine skin. Skin was placed on 96-well plates containing S. typh-lux, imaged (5 min) using a CCD camera, and then completely immersed in PBS, GLY, DMSO, GLY+DMSO in a dose- and time-dependent manner and re-imaged (5 min). The percent of photonic emissions detected (treated or untreated skin relative to no skin controls) was used for analysis. Treatment for 4 h with 50% GLY-PBS and 50:30:20% GLY:DMSO:PBS increased photonic detection compared to untreated skin, 100% PBS, or 30:70% DMSO:PBS. Treatment with 50% GLY in the presence of 20 and 40% DMSO (v/v with PBS) increased photonic detection compared to 50% GLY alone and in the presence of 10% DMSO: 50% GLY (v/v with PBS). Data indicate that GLY and GLY+DMSO are effective optical clearing agents on porcine skin (2-3 mm thick) when treated for 4 h to increase detection of emitted photons. Clearing agents such as GLY have the potential to minimize effects of porcine skin tissue as one of the photon transmittance barriers (i.e., skin, fat, muscle, and visceral tissues) in vivo.

HORSE SPECIES SYMPOSIUM: A novel approach to monitoring pathogen progression during uterine and placental infection in the mare using bioluminescence imaging technology and lux-modified bacteria1,2

“…The use of bioluminescent bacteria, such as E. coli-pAKl-lux or E. coZi!-Xenl4, coupled with bioluminescence imaging technologies, may represent an efficient model for achieving a greater understanding of the pathogenesis of uterine and placental infections in domestic species. Some studies have established positive relationships between bioluminescent signals and bacterial populations for gram-negative bacteria (r = 0.99; Kadurugamuwa et al, 2005), whereas other investigators have described and validated bioluminescent bacterial models, both in vitro (Moulton et al, 2006(Moulton et al, , 2009c and ex vivo (Moulton et al, 2009a,d;Curbelo et al, 2010). More recently, studies have reported the use of bioluminescent reporters and biophotonic imaging technology to study mechanisms of pathogenesis of such pathogens as Staphylococcus aureus (Kuklin et al, 2003), Aspergillus fmnigatus (Brock et al, 2008), Listeria monocytogenes (Konjufca and Miller, 2009), E. coli (Foucault et al, 2010), and Mycobacterium tuberculosis (Andreu et al, 2010); however, these studies have focused on the use of the mouse model.…”

Short-term and long-term effects of optical clearing agents on blood vessels in chick chorioallantoic membrane

“…[1][2][3]5 Higher-concentrated glycerol ͑75%, 100%͒ and glucose ͑40%͒ have great optical clearance of tissue, but the less-concentrated glucose solutions ͑35% to 20%, 10 L͒ have slight clearance. 24,25 Moreover, the agents have also been applied to study short-term effects on blood vessels.…”

“…The tissue optical clearing technique based on immersion of tissues into optical clearing agents, proposed by Tuchin, [1][2][3][4] can improve the depth to which light penetrates, and currently attracts great attention. This technique, combined with other optical imaging techniques, such as optical coherence tomography ͑OCT͒, 2,3,14 fluorescence imaging, 5,6 microscopy imaging, 7,8 and laser radiation, 9,10 etc. will enhance the capabilities of noninvasive optical diagnosis and therapeutic treatments.…”

“…[19][20][21][22] Glycerol and glucose, the common agents, have been proved to be effective for the optical clearance of tissue. [1][2][3]5 However, some side effects of the two agents on blood vessels have been found. [23][24][25] Cheng et al 23 noticed that 100% glycerol induced a decrease of cerebral blood flow and indicated that it is not a good choice for in vivo clearing although it is not toxic.…”

Quantitative analysis method to evaluate optical clearing effect of skin using a hyperosmotic chemical agent

“…Other people measured optical properties to find out the optical clearing effect [20][21]. Jinhee Yoon Author is with the Department of biomedical Engineering, Yonsei University, South Korea (e-mail: no-fate@yonsei.ac.kr).…”