Research Group of Dr. Jean-Marie Botto, ISP Vincience™

Research Group of Dr. Jean-Marie Botto, ISP Vincience™

The Skin Research Unit team, part of the Materials Science Department at ISP Vincience™. From Left to right: Laurine Bergeron, Florian Labarrade, Alexia Lebleu, Amandine Plantivaux, Christelle Plaza, Jean-Marie Botto, Emilie Court and Ludivine Mur.

The ISP Vincience™, situated in Sophia-Antipolis, France, is part of the International Speciality Products (ISP) group and is directed by Dr. Nouha Domloge. The facility has more than 40 scientists and mostly contains the Skin and Hair Research Centre.

Dr. Jean-Marie Botto has been involved with research at ISP Vincience™ since 2000 and is the Senior Manager of the Materials Science department. The department comprises four teams (Skin Research, Hair Biology, Oral Care and Safety and Alternative Toxicology), and is dedicated to the study of skin, hair and oral physiology and the evaluation of the effects and toxicology of dermatological and cosmetic compounds.

Prior to joining ISP Vincience™, Jean-Marie held a postdoctoral research position, and was an Assistant Professor, in Marseille, France. He also holds a Ph.D. in Molecular Pharmacology and Neurosciences, a postgraduate degree in Molecular Pharmacology and a master's degree in Biochemistry, Endocrinology and Molecular Biology from the University of Nice, France.

Jean-Marie says, “In our research, in parallel with molecular approaches (microarrays, qPCR, stable cell lines, etc.), microscopy, especially epi-fluorescence, is crucial both for the cytological and histological examination of different skin cell types and tissues and for assessing the ultra-structure and physiology of hair”.

“We were looking for a solution which would allow us to better understand and present our data, both qualitatively and quantitatively. We also wanted to present our data both internally, and to our customers, in a more engaging way. For this reason, we chose PerkinElmer’s Volocity® imaging solution. The Volocity software is coupled to a motorized microscope, and complements our group of epi-fluorescence microscopes. We now have access to 3D images which are of confocal quality after deconvolution with the iterative restoration algorithm in Volocity. This allows us to investigate the cytology of the tissue sections in more detail. We have also been able to expand our use of Volocity for images generated with our in vivo reflective confocal imaging system, which we use for clinical assessment. The complementary Volocity products: Acquisition, Visualization, Quantitation and Restoration, offer us a complete, yet modular solution”.

“The ability to manage images acquired on several computer workstations in parallel, via the Imaging License Server (ILS), has improved our workflow, by freeing up the system, post-acquisition, to analyze the images. We are now dealing with our microscopy data much more effectively, and acquiring images of better quality and resolution, in 2D and 3D”.

“Finally, we have recently chosen to use PerkinElmer's TSA™ (Tyramide Signal Amplification) system in order to amplify the signal for our in situ hybridization and immunofluorescence applications. In particular, this technology allows us to reduce the concentration of primary antibodies and to reveal the weak expression signals of protein markers or mRNA without increasing the background noise. As a result of this, we have been able to visualize in fine detail, the localization of respiratory chain proteins within mitochondria”.

“By combining the signal sensitivity of TSA with 2D/3D analysis in Volocity, we now have access to structural information and detailed localization of expression, which is essential for our research, for example, the expression confined to certain discrete nuclear regions, of proteins involved in the regulation of expression of certain genes which are important for cutaneous tissue homeostasis”.

 

Deconvolution using Volocity Restoration - Keratin 15 in the peripheral layer of the external root sheath of hair (x63)Deconvolution of an immuno-stained image showing Keratin 15 in the peripheral layer of the external root sheath of hair (x63). The fluorescent image was deconvolved and 3D rendered using Volocity.

 

 

Tyramide Signal Amplification (TSA) - Basal layer of keratinoctyes of the human epidermisAn immuno-marker specific for the basal layer of keratinoctyes of the human epidermis. The sample was treated either with (right) or without (left) TSA. In the lower images, the nuclei are shown (visualized using DAPI).