PIC-ADN

Paoli Calmettes Institute

Magda Rodrigues

Platform manager

Christophe Lachaud

Platform manager

Lara Lee

Engineer

The Platform of Imaging and Characterization of DNA Damage and Instability (PIC-DNA) offers a photonic microscopy facilty, including widefield, confocal, and structured illumination microscopy, along with a service dedicated to the study of genomic instability. It provides equipment for the analysis of samples, whether fixed or observed in live conditions. Open to CRCM teams and private partners, it also offers training, consulting, and support in image processing.

The Platform of Imaging and Characterization of DNA Damage and Instability is essentially organized around photonic microscopy techniques, including widefield, confocal, and structured illumination microscopy…
It offers a coherent set of equipment enabling various approaches for sample analysis: either on fixed cells or tissue sections labeled with fluorescence or dyes, or on live samples studied dynamically, using fluorescence or phase-contrast techniques.
This platform is shared by all CRCM research teams and is also open to private partners. It offers a range of services including training, consulting, support for data interpretation, protocol optimization, image processing and analysis, technology monitoring, as well as management and organization.
Additionally, it provides a full-service for the study of genomic instability ; from sample preparation and image acquisition to data analysis and figure formatting.
The platform is managed and coordinated by a technical engineer (Magda Rodrigues) with support from an another technical engineer (Lara Lee) which is dedicated to the service for the study of genomic instability.

The platform includes the following equipment:

> A Zeiss LSM 880 confocal microscope equipped with the Airyscan enhancement module.
> A NIKON AX Confocal microscope equipped with a UVA (365nm) FRAP module.
> A multi-point confocal microscope equipped with a spinning disk, FRAP, FLIP, photo-activation system able to operate in TIRF mode.
> A structured illumination microscope (Zeiss Apotome) equipped with a color camera for colored samples and a black and white camera for fluorescence.
> Two time-lapse microscopes (Zeiss-Roper Scientifics and Olympus) for fluorescence.
> A fluorescence microscope equipped for molecular combing.
> A Genomic Vision fluorescence slide scanner.
> Analysis stations for image processing.

The platform is constantly evolving to try to best adapt to the reality of projects and user needs.

Equipment

Confocal LSM 880 + Airyscan

LSM 880 spectral + Airyscan module, 2 PMT + 1GaAsP spectral, motorized inverted stand. Excitation wavelengths one UV diode (405nm), 3 lasers in the visible range (1 argon laser at 458 nm, 488 nm, and 514 nm, a DPSS at 561 nm and a HeNe at 633 nm). Objectives: 10X/NA 0.3; 20X/NA 0.8; 25X/NA 0.8; 40X/NA 1.3; 63X/NA 1.4; 100X/NA 1.4. The microscope is equipped with a environnemental control chamber, enabling live samples to be acquired over time. The spectral analysis module subtracts autofluorescence, or separates fluorochromes with overlapping excitation and emission spectra. The Airyscan module enhances resolution.

illustration of LSM 880 confocal equipment

Confocal AX NIKON + FRAP UV

NIKON AX confocal + FRAP UVA module, 1 HS-PMT + 3 GaAsP, inverted and motorized stand + perfect focus system. Excitation wavelengths: 405 nm, 488 nm, 561 nm, and 633 nm. For the FRAP UVA module: a continuous laser at 360 nm. Objectives: 10X/NA 0.45; 20X/NA 0.75; 25X/NA 0.8; 40X/NA 0.75; 40X/NA 1.15; 40X/NA 1.3; 60X/NA 1.4; 100X/NA 1.4. The microscope is equipped with a environmental -control chamber enabling the acquisition of live samples over time.

illustration of AX NIKON confocal equipment

Confocal Spinning-Disk ZEISS

Inverted and motorized stand ZEISS Axio Observer Z1 + definite-focus system. Spinning disk: Yokogawa CSU-X1A. Double Camera and double emission filter wheel 2 x 6 positions. TIRF mode: 1 Camera. Cameras Photometrics EM-CCD Evolve. Illumination system: ILas 2 system; 4-laser bench (405nm, 491, 561, 642 nm). Objectives: 10X/NA 0.3; 40X/NA 1.3; 63X/NA 1.46; 100X/NA 1.45. The microscope is equipped with a environmental control chamber enabling acquisition of live samples over time. Up to 30 images/second on two fluorescent channels simultaneously in Spinning mode. Working in TIRF (Total Illumination Reflexion Microscopy) 2-Color GFP & mCherry mode. FRAP (Fluorescent Recovery After Photobleaching) system for the analysis of molecular dynamics.

illustration of ZEISS spinning-disk confocal equipment

APOTOME ZEISS microscope

Structured illumination microscope to enhance image contrast and resolution through a grid projection system that eliminates out-of-focus light. XYLIS LED excitation allows acquisition of up to 4 fluorochromes. Z-stack and Tilescan with stitching. Camera ORCA FLASH4 LT+ Hammamatsu and a Color camera for the acquisition of colored samples. Objectives: 10X/NA 0.3; 25X/NA 1.2; 40X/NA 1.3; 63X/NA 1.4; 100X/NA 1.4.

ZEISS Videomicroscope

Wide-field microscope dedicated to time-lapse acquisition. Fluorescence excitation by CoolLed pe800 for acquisition of up to 4 fluorochromes and/or phase contrast. METAMORPH control software : Time-lapse, multiposition, z-stack mosaic. sCMOS BT-fusion camera Hammamatsu. Objectives: 5X/NA 0.15; 10X/NA 0.3; 40X/NA 0.6; 63X/NA 1.4

Olympus Videomicroscope

Wide-field microscope dedicated to time-lapse acquisition. Fluorescence excitation by XYLIS LED for acquisition of up to 4 fluorochromes and/or phase contrast. CellSens control software : Time-lapse, multiposition, z-stack mosaic Camera ORCA FLASH4 LT+ Hammamatsu. Objectives: 4X/NA 0.15; 10X/NA 0.3; 40X/NA 0.6; 63X/NA 1.4

Fluorescence slide scanner

Genomic Vision automatic fluorescence slide scanner. Allows full automatic scanning of up to 12 slides and up to 3 fluorochromes. Lumencor Pco edge 4.2 camera. 6-channel Lumencor LED excitation. Objective ZEISS 40X/NA 0.95. filters: CY2/CY3/CY5.

Featured Publications

08/2023

A clickable melphalan for monitoring DNA interstrand crosslink accumulation and detecting ICL repair defects in Fanconi anemia patient cells. Nucleic Acids Research, 51(15), 7988-8004.

Berrada, S., Martínez-Balsalobre, E., Larcher, L., Azzoni, V., Vasquez, N., Da Costa, M., Abel, S., Audoly, G., Lee, L., Montersino, C., Castellano, R., Combes, S., Gelot, C., Ceccaldi, R., Guervilly, J., Soulier, J., & Lachaud, C.

09/2021

UFMylation of MRE11 is essential for telomere length maintenance and hematopoietic stem cell survival. Science Advances, 7(39).

Lee, L., Oliva, A. B. P., Martinez-Balsalobre, E., Churikov, D., Peter, J., Rahmouni, D., Audoly, G., Azzoni, V., Audebert, S., Camoin, L., Mulero, V., Cayuela, M., Kulathu, Y., Geli, V., & Lachaud, C.

09/2023

PDZ scaffolds regulate extracellular vesicle production, composition, and uptake. Proceedings of the National Academy of Sciences, 120(38).

Castro-Cruz, M., Hyka, L., Daaboul, G., Leblanc, R., Meeussen, S., Lembo, F., Oris, A., Van Herck, L., Granjeaud, S., David, G., & Zimmermann, P.

03/2024

Absence of chromosome axis protein recruitment prevents meiotic recombination chromosome-wide in the budding yeast Lachancea kluyveri. Proceedings of the National Academy of Sciences, 121(12).

Legrand, S., Saifudeen, A., Bordelet, H., Vernerey, J., Guille, A., Bignaud, A., Thierry, A., Acquaviva, L., Gaudin, M., Sanchez, A., Johnson, D., Friedrich, A., Schacherer, J., Neale, M. J., Borde, V., Koszul, R., & Llorente, B.

09/2024

Heterozygous RPA2 variant as a novel genetic cause of telomere biology disorders. Genes & Development.

Kochman, R., Ba, I., Yates, M., Pirabakaran, V., Gourmelon, F., Churikov, D., Lafaille, M., Kermasson, L., Hamelin, C., Marois, I., Jourquin, F., Braud, L., Bechara, M., Lainey, E., Nunes, H., Breton, P., Penhouet, M., David, P., Géli, V., & Coulon, S.

10/2023

Downregulation of stromal syntenin sustains AML development. EMBO Molecular Medicine, 15(11).

Leblanc, R., Ghossoub, R., Goubard, A., Castellano, R., Fares, J., Camoin, L., Audebert, S., Balzano, M., Bou-Tayeh, B., Fauriat, C., Vey, N., Garciaz, S., Borg, J., Collette, Y., Aurrand-Lions, M., David, G., & Zimmermann, P.

02/2024

mTert induction in p21-positive cells counteracts capillary rarefaction and pulmonary emphysema. EMBO Reports, 25(3), 1650-1684

Lipskaia, L., Breau, M., Cayrou, C., Churikov, D., Braud, L., Jacquet, J., Born, E., Fouillade, C., Curras-Alonso, S., Bauwens, S., Jourquin, F., Fiore, F., Castellano, R., Josselin, E., Sánchez-Ferrer, C., Giovinazzo, G., Lachaud, C., Gilson, E., Flores, I., & Géli, V.

04/2022

Discovery of Small-Molecule Inhibitors of the PTK7/β-Catenin Interaction Targeting the Wnt Signaling Pathway in Colorectal Cancer. ACS Chemical Biology, 17(5), 1061-1072.

Ganier, L., Betzi, S., Derviaux, C., Roche, P., Dessaux, C., Muller, C., Hoffer, L., Morelli, X., & Borg, J.

04/2024

A vertebrate Vangl2 translational variant required for planar cell polarity. Journal of Biological Chemistry, 300(4), 106792.

Walton, A., Thomé, V., Revinski, D., Marchetto, S., Puvirajesinghe, T. M., Audebert, S., Camoin, L., Bailly, E., Kodjabachian, L., & Borg, J.

09/2022

The serine/threonine kinase MINK1 directly regulates the function of promigratory proteins. Journal of Cell Science, 135(17).

Daulat, A. M., Wagner, M. S., Audebert, S., Kowalczewska, M., Ariey-Bonnet, J., Finetti, P., Bertucci, F., Camoin, L., & Borg, J.

02/2021

Syntenin-knock out reduces exosome turnover and viral transduction. Scientific Reports, 11(1).

Kashyap, R., Balzano, M., Lechat, B., Lambaerts, K., Egea-Jimenez, A. L., Lembo, F., Fares, J., Meeussen, S., Kügler, S., Roebroek, A., David, G., & Zimmermann, P.

All our publications