Formation of hemoglobin photoproduct is responsible for two-photon and single photon-excited fluorescence of red blood cells

Shirshin, Evgeny A.; Yakimov, Boris P.; Rodionov, Sergei A.; Omelyanenko, N. P.; Priezzhev, Alexander V.; Фадеев, В. В.; Lademann, Juergen; Darvin, Maxim E.

doi:10.1088/1612-202x/aac003

Cited by 22 publications

(16 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…They are localized in the proximity of blood capillaries, which are visible in dark red in the TPE-FLIM images (Fig. 6b, d, f) due to ultrafast fluorescence decay of Hb photoproducts 59 . The presence of cytoplasmic granules is a specific hallmark feature of MCs.…”

Section: Discussionmentioning

confidence: 99%

In vivo non-invasive staining-free visualization of dermal mast cells in healthy, allergy and mastocytosis humans using two-photon fluorescence lifetime imaging

Kröger

Nikolaev

Shirshin

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Mast cells (MCs) are multifunctional cells of the immune system and are found in skin and all major tissues of the body. They contribute to the pathology of several diseases including urticaria, psoriasis, atopic dermatitis and mastocytosis where they are increased at lesional sites. Histomorphometric analysis of skin biopsies serves as a routine method for the assessment of MC numbers and their activation status, which comes with major limitations. As of now, non-invasive techniques to study MCs in vivo are not available. Here, we describe a label-free imaging technique to visualize MCs and their activation status in the human papillary dermis in vivo. This technique uses two-photon excited fluorescence lifetime imaging (TPE-FLIM) signatures, which are different for MCs and other dermal components. TPE-FLIM allows for the visualization and quantification of dermal MCs in healthy subjects and patients with skin diseases. Moreover, TPE-FLIM can differentiate between two MC populations in the papillary dermis in vivo—resting and activated MCs with a sensitivity of 0.81 and 0.87 and a specificity of 0.85 and 0.84, respectively. Results obtained on healthy volunteers and allergy and mastocytosis patients indicate the existence of other MC subpopulations within known resting and activated MC populations. The developed method may become an important tool for non-invasive in vivo diagnostics and therapy control in dermatology and immunology, which will help to better understand pathomechanisms involving MC accumulation, activation and degranulation and to characterize the effects of therapies that target MCs.

show abstract

Section: Discussionmentioning

confidence: 99%

In vivo non-invasive staining-free visualization of dermal mast cells in healthy, allergy and mastocytosis humans using two-photon fluorescence lifetime imaging

Kröger

Nikolaev

Shirshin

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…In [51] the authors propose the usage of green endogenous AF as a real-time quantification of cellular senescence in human mesenchymal stromal cells based on label-free flow cytometry analysis and the involvement of lipofuscin in the AF increase was discussed. In our previous work it was shown that the appearance of the fluorescence signal in the living cells can be governed by the formation of photoproducts: for instance, the irradiation of erythrocytes with 760 nm femtosecond laser pulses led to the formation of hemoglobin photoproducts which were characterized by a broad excitation spectrum [52].…”

Section: Discussionmentioning

confidence: 99%

The Oxidation-Induced Autofluorescence Hypothesis: Red Edge Excitation and Implications for Metabolic Imaging

et al. 2020

Self Cite

View full text Add to dashboard Cite

Endogenous autofluorescence of biological tissues is an important source of information for biomedical diagnostics. Despite the molecular complexity of biological tissues, the list of commonly known fluorophores is strictly limited. Still, the question of molecular sources of the red and near-infrared excited autofluorescence remains open. In this work we demonstrated that the oxidation products of organic components (lipids, proteins, amino acids, etc.) can serve as the molecular source of such red and near-infrared excited autofluorescence. Using model solutions and cell systems (human keratinocytes) under oxidative stress induced by UV irradiation we demonstrated that oxidation products can contribute significantly to the autofluorescence signal of biological systems in the entire visible range of the spectrum, even at the emission and excitation wavelengths higher than 650 nm. The obtained results suggest the principal possibility to explain the red fluorescence excitation in a large class of biosystems—aggregates of proteins and peptides, cells and tissues—by the impact of oxidation products, since oxidation products are inevitably presented in the tissue. The observed fluorescence signal with broad excitation originated from oxidation products may also lead to the alteration of metabolic imaging results and has to be taken into account.

show abstract

“…Further studies revealed that single (in the 400-500 nm range) and two-photon excited fluorescence of RBC is due to the formation of Hb photoproduct [29]. As a consequence, fluorescence intensity of RBC suspension and Hb aqueous solution increases linearly with the irradiation time [29].…”

Section: Autofluorescence Of Red Blood Cellsmentioning

confidence: 96%

Label-free characterization of white blood cells using fluorescence lifetime imaging and flow-cytometry: molecular heterogeneity and erythrophagocytosis [Invited]

Yakimov¹,

Gogoleva²,

Semenov³

et al. 2019

Biomed. Opt. Express

View full text Add to dashboard Cite

Blood cell analysis is one of the standard clinical tests. Despite the widespread use of exogenous markers for blood cell quantification, label-free optical methods are still of high demand due to their possibility for in vivo application and signal specific to the biochemical state of the cell provided by native fluorophores. Here we report the results of blood cell characterization using label-free fluorescence imaging techniques and flow-cytometry. Autofluorescence parameters of different cell types -white blood cells, red blood cells, erythrophagocytic cells -are assessed and analyzed in terms of molecular heterogeneity and possibilities of differentiation between different cell types in vitro and in vivo.

show abstract

Formation of hemoglobin photoproduct is responsible for two-photon and single photon-excited fluorescence of red blood cells

Cited by 22 publications

References 35 publications

In vivo non-invasive staining-free visualization of dermal mast cells in healthy, allergy and mastocytosis humans using two-photon fluorescence lifetime imaging

In vivo non-invasive staining-free visualization of dermal mast cells in healthy, allergy and mastocytosis humans using two-photon fluorescence lifetime imaging

The Oxidation-Induced Autofluorescence Hypothesis: Red Edge Excitation and Implications for Metabolic Imaging

Label-free characterization of white blood cells using fluorescence lifetime imaging and flow-cytometry: molecular heterogeneity and erythrophagocytosis [Invited]

Contact Info

Product

Resources

About