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Autofluorescence Spectroscopy for Monitoring Metabolism in Animal Cells and Tissues.
Methods Mol Biol
February 2018
Department of Biology and Biotechnology "Lazzaro Spallanzani", University of Pavia, Pavia, Italy.
Excitation of biological substrates with light at a suitable wavelength can give rise to a light emission in the ultraviolet (UV)-visible, near-infrared (IR) spectral range, called autofluorescence (AF). This is a widespread phenomenon, ascribable to the general presence of biomolecules acting as endogenous fluorophores (EFs) in the organisms of the whole life kingdom. In cytochemistry and histochemistry, AF is often an unwanted signal enhancing the background and affecting in particular the detection of low signals or rare positive labeling spots of exogenous markers.
View Article and Find Full Text PDFAutofluorescence properties of murine embryonic stem cells during spontaneous differentiation phases.
Lasers Surg Med
November 2013
IGM-CNR and Department of Biology & Biotechnology, University Pavia, Pavia, 27100, Italy.
Background And Objective: The autofluorescence (AF) analysis allows in vivo, real-time assessment of cell functional activities, depending on the presence of biomolecules strictly involved in metabolic reactions and acting as endogenous fluorophores. Pluripotent stem cells during differentiation are known to undergo changes in their morphofunctional properties, with particular reference to bioenergetic metabolic signatures involving endogenous fluorophores such as NAD(P)H, flavins, lipofuscin-like lipopigments. Since the development of regenerative therapies based on pluripotent cells requires a careful monitoring of the successful maturation into the desired phenotype, aim of our work is to evaluate the AF potential to assess the differentiation phases in a murine stem cell model.
View Article and Find Full Text PDFChanges in the activity of defense mechanisms against induced acidosis during meiotic maturation in mouse oocytes.
Theriogenology
April 2011
Department of Physiology, Faculty of Medicine, University of Cukurova, Adana, Turkey.
Previous work has indicated that although activity of the HCO₃⁻/Cl⁻exchanger (AE), which regulates intracellular alkalosis, is high in the germinal vesicle (GV) stage, the oocyte is inhibited as it progresses through meiotic maturation. In this study, we aimed to investigate the defense mechanisms against acidosis during the meiotic maturation stages. Intracellular pH (pH(i)) was recorded using a microspectrofluorometric technique, and Na(+)/H(+) (NHE) and Na(+)-dependent HCO₃⁻/Cl⁻ exchanger (NDCBE) activity were determined by measuring the recovery rate from induced acidosis.
View Article and Find Full Text PDFHuman liver autofluorescence: an intrinsic tissue parameter discriminating normal and diseased conditions.
Lasers Surg Med
July 2010
Histochemistry and Cytometry Section, IGM-CNR, Pavia 27100, Italy.
Background And Objective: Autofluorescence (AF) emission is an intrinsic parameter that can provide real-time information on morpho-functional properties of biological tissue, being strictly related with their biochemical composition and structural organization. The diagnostic potentials of AF-based techniques have been investigated on normal, fibrotic, and steatotic liver tissues, in reference to histological features as evidenced by specific histochemical stainings.
Materials And Methods: AF emission under excitation at 366 nm has been examined on cryostatic tissue sections obtained from biopsies collected during surgical operation, by means of fluorescence imaging and microspectrofluorometric techniques.
The developmental basis of an evolutionary diversification of female gametophyte structure in Piper and Piperaceae.
Ann Bot
April 2009
University of Colorado, Department of Ecology and Evolutionary Biology, Boulder, CO 80309, USA.
Background And Aims: Fritillaria-type female gametophyte development is a complex, yet homoplasious developmental pattern that is interesting from both evolutionary and developmental perspectives. Piper (Piperaceae) was chosen for this study of Fritillaria-type female gametophyte development because Piperales represent a 'hotspot' of female gametophyte developmental evolution and have been the subject of several recent molecular phylogenetic analyses. This wealth of phylogenetic and descriptive data make Piper an excellent candidate for inferring the evolutionary developmental basis for the origin of Fritillaria-type female gametophytes.
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