The nature of the chromophore responsible for naturally occurring fluorescence in mouse skin

doi:10.1016/1011-1344(88)85051-6

Journal of Photochemistry and Photobiology B: Biology

Volume 2, Issue 3, November 1988, Pages 313-320

https://doi.org/10.1016/1011-1344(88)85051-6 Get rights and content

Abstract

Normal mouse skin has a prominent fluorescence peak at 674 nm. Fluorescence emission and fluorescence excitation spectroscopy, carried out both in vitro and in vivo, led to the conclusion that the chromophore(s) responsible for this naturally occurring fluorescence is/are pheophorbide a and/or pheophytin a, degradation products of chlorophyll a that are derived from the mouse food.

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Citation Excerpt :
The emission band around 510 nm is characteristic of lipids from the lamellar granules in the keratinocyte, substances such as ceramides, fatty acids, and cholesterol that comprises the intracellular spaces of the stratum corneum, elastin, and mainly flavins from epidermal cells and hair follicles [34]. The other two bands at 633 and 668 nm can be associated with porphyrins emission and degradation products of chlorophyll derived from food [27,35]. The spectrum of ALA-induced PpIX after 1 h of incubation also showed the 633 nm band strongly suggesting that the fluorescence signal is mainly related to PpIX (Fig. 3B).
Ultraviolet (UV) radiation may induce skin alterations as observed in photoaging. Some recognized modifications are epidermal hyperplasia, amorphous deposition of degraded elastic fibers and reduction in the number of collagen fibers. They alter the tissue biochemical properties that can be interrogated by steady state fluorescence spectroscopy (SSFS). In this study, we monitored the changes in endogenous fluorescence emission from hairless mice skin during a protocol of photoaging using UVB irradiation. To perform the fluorescence spectroscopy, it was used a violet laser (408 nm) to induce the native fluorescence that is emitted in the visible range. Under 408 nm excitation, the emission spectrum showed bands with peaks centered around 510, 633 and 668 nm for irradiated and control groups. A relative increase of the fluorescence at 633 nm emission on the flank was observed with time when compared to the ventral skin at the same animal and the non-irradiated control group. We correlated the emission at 633 nm with protoporphyrin IX (PpIX), and our hypothesis is that the PpIX metabolism in the photoaged and aged skin are different. PpIX fluorescence intensity in the photoaged skin is higher and more heterogeneous than in the aged skin. Notwithstanding, more spectroscopic and biochemistry studies investigating the 510 and 633 nm emission are needed to confirm this hypothesis.
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Chlorophylls occur abundantly in food plants and show bright emission bands at long-wavelength regions (∼675 and ∼720 nm) compared to the autofluorescence of animal organs and peritoneal fluids. The use of these emissions as biomarkers for monitoring bowel perforation with a modality that does not involve synthetic contrast agents seems promising. To validate this, we measured the fluorescence spectra of rat organs, human peritoneal and intestinal fluids, and human intestinal fluids diluted with physiological saline. The developed technique showed a high detection sensitivity (∼50 ppm) under irrigation for abdominal surgery, highlighting the potential of this tool in the surgical setting.
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Nonlinear optical imaging (NLOI) has emerged to be a promising tool for bio-medical imaging in recent times. Among the various applications of NLOI, its utility is the most significant in the field of pre-clinical and clinical cancer research. This review begins by briefly covering the core principles involved in NLOI, such as two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG). Subsequently, there is a short description on the various cellular components that contribute to endogenous optical fluorescence. Later on the review deals with its main theme – the challenges faced during label-free NLO imaging in translational cancer research. While this review addresses the accomplishment of various label-free NLOI based studies in cancer diagnostics, it also touches upon the limitations of the mentioned studies. In addition, areas in cancer research that need to be further investigated by label-free NLOI are discussed in a latter segment. The review eventually concludes on the note that label-free NLOI has and will continue to contribute richly in translational cancer research, to eventually provide a very reliable, yet minimally invasive cancer diagnostic tool for the patient.
On the autofluorescence of fingermarks
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Citation Excerpt :
Demetallation of chlorophyll leads to the formation of pheophytin. Pheophorbide A is a dephytylated pheophytin [9,12,18] which is known to reach the skin, sometimes leading to phototoxicity [12,18]. In MP1 their Rf-value differed, 0.97 and 0.72 respectively.
The autofluorescence of fingermarks is used for their detection. The components responsible for this autofluorescence are largely unknown. Thin layer chromatography and fluorescence spectroscopy were used to identify autofluorescent components and evaluate their forensic value. Based on our results, tryptophan is hypothesized to be a major contributor to the autofluorescence when part of peptides or proteins, id est, not in its free form. Part of the autofluorescence could be assigned to a kynurenine derivative. Pheophorbide A, a metabolite of chlorophyll, is inferred as a red fluorescent fingermark component. Chlorophyll is a plant pigment which implies that dietary information can potentially be retrieved from fingermarks.
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Journal of Photochemistry and Photobiology B: Biology

Abstract

References (17)

Pyrrole pigments, isoprenoid compounds and phenolic plant constituents

Visible absorption and fluorescence of chlorophyll and its aggregates in solution

Time-resolved fluorescence spectroscopy of hematoporphyrin, mesoporphyrin, pheophorbide a and chlorin e₆ in ethanol and aqueous solution

J. Photochem. Photobiol., B: Biol.

Laser excited fluorescence of HpD for diagnosis of cancer

IEEE J. Quantum Electron

Fluorescence of hematoporphyrin-derivative for detection and characterization of tumors

A new diagnostic system for malignant tumors using hematoporphyrin derivative, laser photoradiation and a spectroscope

Non-invasive technique for obtaining fluorescence excitation and emission spectra in vivo

Photochem. Photobiol.

Biological activities of phthalocyanines — V. Photodynamic therapy of EMT-6 mammary tumors in mice with sulfonated phthalocyanines

Photochem. Photobiol.

Cited by (79)

Fluorescence spectroscopy in the visible range for the assessment of UVB radiation effects in hairless mice skin

Contrast agent free detection of bowel perforation using chlorophyll derivatives from food plants

Advances and challenges in label-free nonlinear optical imaging using two-photon excitation fluorescence and second harmonic generation for cancer research

On the autofluorescence of fingermarks

Minimizing near-infrared autofluorescence in preclinical imaging with diet and wavelength selection

Photoluminescence in mammal fur: 111 years of research