Technologies
Our development team has more than fifteen years of experience in designing and building advanced imaging hardware and software. We work hand in hand with our customers in precisely identifying the specific requirements of their research applications and develop customized imaging solutions that best suit their needs. Having worked ourselves with commercially available microscopy systems, we fully understand what the term “user-friendly” means and we design image acquisition software that can be rightfully described as such. We provide expert consultation in building modular systems. This means that customers actively participate in selecting the key components of their own systems within a wide variety of qualities and prices. This also implies that our systems are dynamic, they can be rapidly and easily adapted as new technologies, better hardware or more advance software are being developed or to better accommodate to the changing demands of your research projects.
What technologies can we help you implement in your laboratories?
Selective Plane Illumination Microscopy (SPIM)
SPIM, Ultramicroscopy, or in general Laser Sheet Microscopy techniques enable fast high-resolution imaging of large, optically-cleared, three-dimensional specimens. In SPIM, whole organs, thick tissue samples or organotypic or 3D cellular cultures are illuminated using a thin sheet of light that scans the specimens along one axis while emitted light is collected along a perpendicular optical axis, ensuring that the detection objective focal plane coincides with the sheet of light. In the past decade different modalities of SPIM have been developed that vary depending on the configuration of sources and cameras, and on how the sheet is generated to reduce scattering and increase speed. We develop Laser Sheet Microscopy set-ups in all configurations (single-armed, double-sided illumination, fast-acquisition for in-vivo, multiview, etc.)
Advantages
Low photo-toxicity and photo bleaching, high acquisition speeds, sensitive detection, good light penetration, 3D resolution.
References using SPIM and software of 4D-nature
- Alicia Arranz, Di Dong, Shouping Zhu, Markus Rudin, Christos Tsatsanis, Jie Tian, and Jorge Ripoll, “Helical optical projection tomography,” Opt. Express 21, 25912-25925 (2013). In this paper we combined SPIM with OPT (see below) for imaging large samples.
Optical projection Tomography (OPT)
OPT is a mesoscopic technique (Science 296 (5567): 541–5. April 2002) that is used to acquire panoramic 3D microscopy images of samples considered too big for conventional microscopy techniques (1-10mm). In OPT light is projected through a whole specimen treated with optical clearing agents, and the transmitted light is collected as the object rotates. Using backprojection algorithms, a 3D image of the specimen is reconstructed from the series of images acquired. OPT is rapidly becoming a widely used technique in biomedical research and has been employed so far to visualize fixed specimens such as whole mouse embryos, pancreas and peripheral lymph nodes among other (see for example, Kumar V et al, Front Immunol. 2012;3:282).
Advantages
Imaging of large specimens, isotropic resolution.
Schematics of a combined OPT-SPIM setup capable of performing Helical OPT. Helical OPT is a new technique developed by members of our team for imaging large elongated samples.
References using OPT and software of 4D-nature:
- Alicia Arranz, Di Dong, Shouping Zhu, Markus Rudin, Christos Tsatsanis, Jie Tian, and Jorge Ripoll, “Helical optical projection tomography,” Opt. Express 21, 25912-25925 (2013). In this paper we combined SPIM (see SPIM) with OPT for imaging large samples.
- Rieckher M, Birk UJ, Meyer H, Ripoll J, Tavernarakis N (2011) Microscopic Optical Projection Tomography In Vivo. PLoS ONE 6(4): e18963. doi:10.1371/journal.pone.0018963. Note: In this paper we make use of OPT to image C. elegans in-vivo, being the first time OPT is used on live microscopic specimens.
- Heiko Meyer, Alex Darrell, Athanasios Metaxakis, Charalambos Savakis and Jorge Ripoll, “Optical Projection Tomography for In-Vivo Imaging of Drosophila melanogaster”, Microscopy and Analysis, Sept. (2008).
- D. Dong, S. Zhu, C. Qin, V. Kumar, J. V Stein, S. Oehler, C. Savakis, J. Tian, and J. Ripoll. “Automated Recovery of the Center of Rotation in Optical Projection Tomography in the Presence of Scattering.” IEEE Trans. Inf. Tech. in Biomed (Sept. 2012).
- S. Zhu, D. Dong, U. J. Birk, M. Rieckher, N. Tavernarakis, X. Qu, J. Liang, J. Tian, and J. Ripoll. Automated motion correction for in vivo optical projection tomography.” IEEE Trans Med Imaging 31.7 (July 2012), pp. 1358-71.
Fluorescence Molecular Tomography (FMT)
FMT is a tomographic technique that enables in-vivo 3D quantitative images of near infra-red fluorescent probes and proteins in deep tissues. FMT has emerged as a key non-invasive imaging technology in biomedical and preclinical research, due to the fact that it is high-throughput and it employs very specific activatable fluorescent probes or fluorescent proteins. It is based on modeling light propagation in tissues through the diffusion approximation in order to solve what is termed an “inverse problem” to recover the 3D spatial distribution of flurophore concentration.
Advantages
Non-invasive, in vivo deep tissue imaging, high sensitivity and specificity.
FMT Setup in reflectance mode, detailing the use of multiwavelength excitation and detection for multispectral FMT imaging
References using FMT and software of 4D-nature
- Abraham Martin, Juan Aguirre , Ana Sarasa, Anikitos Garofalakis, Heiko Meyer, Clio Mamalaki, Jorge Ripoll and Anna M. Planas, “Imaging Changes in Lymphoid Organs In Vivo after Brain Ischemia with Three-Dimensional Fluorescence Molecular Tomography in Transgenic Mice Expressing Green Fluorescent Protein in T Lymphocytes”, Mol. Imag. Jul-Aug;7(4):157-67 (2008). Note: In this paper we make use of FMT to follow the changes of GFP expressing T-Lymphocytes in 3D in-vivo after brain ischemia.
- M. Simantiraki, R. Favicchio, S. Psycharakis, G. Zacharakis and J. Ripoll, “Multispectral unmixing of fluorescence molecular tomography data”, J. of Inn. Opt. Health Sci. Vol. 2(4), 353–364 (2009).
- F. Stuker, C. Baltes, K. Dikaiou, D. Vats, L. Carrara, E. Charbon, J. Ripoll, and M. Rudin. Hybrid small animal imaging system combining magnetic resonance imaging with fluorescence tomography using single photon avalanche diode detectors”. IEEE Trans Med Imaging 30.6 (2011), pp. 1265-1273. Note: In this work FMT was used in combination with MRI, making use of an FMT and software developed by members of the 4D-Nature team.
- Zacharakis, G., Favicchio, R., Simantiraki, M. and Ripoll, J. (2011). Spectroscopic detection improves multi-color quantification in fluorescence tomography. Biomedical Optics Express, 2(3), 431–9. doi:10.1364/BOE.2.000431. Note: In this paper FMT is used for multispectral imaging, making use of volumetric multispectral unmixing techniques.
- R. Favicchio, G. Zacharakis, K. Oikonomaki, A. Zacharopoulos, C. Mamalaki, and J. Ripoll. Kinetics of T-cell receptor-dependent antigen recognition determined in vivo by multi-spectral normalized epiuorescence laser scanning”. Journal of Biomedical Optics 17.7 (2012), p. 076013.
- A. Zelmer, P. Carroll, N. Andreu, K. Hagens, J. Mahlo, N. Redinger, B. D.Robertson, S. Wiles, T. H. Ward, T. Parish, J. Ripoll, G. J. Bancroft, and U. E. Schaible. “A new in vivo model to test anti-tuberculosis drugs using fluorescence imaging.” The Journal of antimicrobial chemotherapy 67.8 (Aug. 2012), pp. 1948-60.
- A. Arranz, A. Androulidaki, B. Mol, E. Tsentelierou, E. N. Stathopoulos, C. Tsatsanis, and J. Ripoll. “Intravital spectral imaging as a tool for accurate measurement of vascularization in mice.” Journal of angiogenesis research 2.1 (Jan. 2010), p. 22.