Breunig H.G.,Saarland University |
Breunig H.G.,JenLab GmbH |
Uchugonova A.,Saarland University |
Uchugonova A.,JenLab GmbH |
And 3 more authors.
Scientific Reports | Year: 2015
Optoporation, the permeabilization of a cell membrane by laser pulses, has emerged as a powerful non-invasive and highly efficient technique to induce transfection of cells. However, the usual tedious manual targeting of individual cells significantly limits the addressable cell number. To overcome this limitation, we present an experimental setup with custom-made software control, for computer-automated cell optoporation. The software evaluates the image contrast of cell contours, automatically designates cell locations for laser illumination, centres those locations in the laser focus, and executes the illumination. By software-controlled meandering of the sample stage, in principle all cells in a typical cell culture dish can be targeted without further user interaction. The automation allows for a significant increase in the number of treatable cells compared to a manual approach. For a laser illumination duration of 100ms, 7-8 positions on different cells can be targeted every second inside the area of the microscope field of view. The experimental capabilities of the setup are illustrated in experiments with Chinese hamster ovary cells. Furthermore, the influence of laser power is discussed, with mention on post-treatment cell survival and optoporation-efficiency rates.
Uchugonova A.,Saarland University |
Uchugonova A.,JenLab GmbH
Journal of Biomedical Optics | Year: 2017
The multiphoton fluorescence lifetime imaging tomograph MPTflex with its flexible 360-deg scan head, articulated arm, and tunable femtosecond laser source was employed to study induced pluripotent stem cell (iPS) cultures. Autofluorescence (AF) lifetime imaging was performed with 250-ps temporal resolution and submicron spatial resolution using time-correlated single-photon counting. The two-photon excited AF was based on the metabolic coenzymes NAD(P)H and flavin adenine dinucleotide/flavoproteins. iPS cells generated from mouse embryonic fibroblasts (MEFs) and cocultured with growth-arrested MEFs as feeder cells have been studied. Significant differences on AF lifetime signatures were identified between iPS and feeder cells as well as between their differentiating counterparts. © 2017 Society of Photo-Optical Instrumentation Engineers (SPIE).
Agency: European Commission | Branch: FP7 | Program: CP | Phase: ICT-2011.3.5 | Award Amount: 13.40M | Year: 2012
Biophotonics offers low-cost, non-invasive, accurate, rapid alternatives to conventional diagnostic methods and has the potential to address medical needs with early detection and to reduce the cost of healthcare. FAMOS will develop a new generation of light sources with step-changes in performance beyond the state-of-the-art to radically transform biophotonic technologies for point-of-care diagnosis and functional imaging. This will enable optical diagnostics with superior sensi-tivity, specificity, reliability and clinical utility at reduced cost, heralding an imaging renaissance in Europe.FAMOS addresses optical imaging from molecular over (sub)cellular to individual organs, with no gap in the arsenal of diagnostic tools for medical end-users. The world-class multidisciplinary FA-MOS team of 7 leading academic institutions and 10 top SMEs has unique complementary knowledge of optical coherence tomography, adaptive optics, photoacoustic tomography, coherent anti-stokes Raman scattering, multiphoton tomography as well as swept-source, diode-pumped ultrafast and tuneable nanosecond pulse lasers. Combinations of some techniques will offer multi-modal solutions to diagnostic needs that will exploit and enhance the benefits of each modality. FAMOS technologies have wide applicability, but our specific focus is on diagnosis in ophthalmol-ogy and oncology. Partnerships with leading innovative clinical users will enable preclinical evalua-tion.The objectives of FAMOS are:\tDevelop new light sources with a step-change in performance (2-3 times more compact and up to 3-4 times cheaper diode pumped Ti:sapphire, 4-10 times faster swept sources and tuneable nanosecond pulse sources)\tIntegrate these with optical imaging for a step-change in diagnosis (2-5 times better resolution cellular retinal imaging with more than 10 times larger field of view, up to 10 times enhanced penetration single source subcellular morphologic imaging, increased selectivity of intrinsic mo-lecular sensing as well as several frames per second deep tissue functional tomography\tPerform preclinical studies to demonstrate novel or improved ophthalmic and skin cancer diag-nosis establishing novel biomarkers (melanocyte shape, NADPH, melanin concentration, Hb/HbO2 as well as lipid, water and DNA/RNA concentration)\tEnable exceptional commercial opportunities for SMEs\tProvide state-of-the-art academic training
Breunig H.G.,JenLab GmbH |
Studier H.,JenLab GmbH |
Konig K.,JenLab GmbH |
Konig K.,Saarland University
Optics Express | Year: 2010
In vivo multiphoton tomography with a wavelength-tunable femtosecond laser has been performed to investigate the autofluorescence intensity of major endogenous fluorophores of human skin in dependence on the excitation wavelength. In high-resolution multiphoton images of different skin layers, clear trends were found for fluorophores like keratin, NAD(P)H, melanin as well as for the elastin and collagen networks. The analysis of the measurements is supplemented by additional measurements of fluorescence lifetime imaging and signal-decay curves by time-correlated single-photon counting. © 2010 Optical Society of America.
Konig K.,Saarland University |
Konig K.,JenLab GmbH |
Uchugonova A.,Saarland University |
Gorjup E.,Fraunhofer Institute for Biomedical Engineering
Microscopy Research and Technique | Year: 2011
Long-term high-resolution multiphoton imaging of nonlabeled human salivary gland stem cell spheroids has been performed with submicron spatial resolution, 10.5-nm spectral resolution, and picosecond temporal resolution. In particular, the two-photon-excited coenzyme NAD(P)H and flavins have been detected by time-correlated single photon counting (TCSPC). Stem cells increased their autofluorescence lifetimes and decreased their total fluorescence intensity during the adipogenic-differentiation process. In addition, the onset of the biosynthesis of lipid vacuoles was monitored over a period of several weeks in stem-cell spheroids. Time-resolved multiphoton autofluorescence imaging microscopes may become a promising tool for marker-free stem-cell characterization and cell sorting. © 2010 Wiley Periodicals, Inc.
JenLab GmbH | Date: 2016-02-09
A method and an apparatus for reprogramming living cells without using viruses. In that method a cocktail comprising at least two transcription factors and a microRNA is transfected into the interior of at least one cell in order to convert this cell into iPS cells or into another type of cell, by storing the cells to be converted in an aqueous environment of the cocktail without viral carriers and focusing a femtosecond laser in a laser scanning microscope with a numerical aperture between 0.9 and 1.5 on a cell membrane of the cell to be reprogrammed and controlling the position of the focus. The exposure period and laser power for the optical treatment of the cell such that the focus depending on the pulse repetition frequency with an output between 7 mW and 100 mW generates a transient small-pore hole with a size up to 500 nm.
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: HEALTH-2007-1.2-1 | Award Amount: 5.23M | Year: 2008
The incidence of skin cancer in Europe, US, and Australia is rising rapidly. One in five will develop some form of skin cancer during the lifetime. A person has a 1:33 chance to develop melanoma, the most aggressive skin cancer. Melanoma is the second most common cancer in women aged 20-29, and the sixth most common cancer in men and women. In 2007, more than 1 million new cases will be diagnosed in the US alone. About 90% of skin cancers are caused by ultraviolet (UV) sun light. The World Health Organization estimates that 60,000 people will die this year from too much sun: 48,000 from melanoma and 12,000 from other skin cancer. A significant improvement of the current diagnostic tools of dermatologists is required in order to identify dermal disorders at a very early stage as well as to monitor directly the effects of treatment. We suggest within this proposal the development of a non-invasive multimodal hybrid imaging system with the capability to perform non-invasive high resolution three-dimensional clinical (i) two-photon imaging with time-correlated single photon detection, (ii) autofluorescence lifetime imaging, (iii) high-frequency acoustical imaging with novel miniaturized multiple detector arrays, and (iv) optoacoustical imaging using ultrashort near infrared (NIR) laser pulses. This novel multimodal approach will provide a wide-field acoustic/optoacoustic view with quantitative depth information of the dermatological lesion as well as a close optical look into particular intratissue compartments with quantitative hyperspectral information and subcellular resolution. A successful project will provide a novel unique tool for early diagnosis and treatment control of skin cancer and skin disease and will significantly contribute to the improvement of the European health care system.
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: HEALTH-2007-1.2-1;HEALTH-2007-1.2-2 | Award Amount: 7.09M | Year: 2008
The aim of FUN OCT is to expand the non-invasive optical biopsy capability of optical coherence tomography (OCT) and combination of OCT with multiphoton tomography (MT) to develop novel functional capabilities hereby enabling morphofunctional performance, i.e., the fusion of anatomic and functional imaging at the cellular resolution level. These methodologies will enable unprecedented non-invasive detection of depth resolved physiological, metabolic as well as molecular specific tissue information, i.e., forming a novel, powerful medical imaging platform. This novel platform fills an important gap left by todays medical imaging technology. The hypothesis is that the combination of cellular resolution, real time imaging of morphology and depth resolved tissue function could enable a major step forward in early cancer diagnosis and in the early detection of retinal pathologies that are world wide leading causes of blindness. This is accomplished due to a synergistic effect from joining complementary international expertise in the fields of laser sources, OCT, MT and beam delivery system technology. The consortium comprises 6 research groups and 2 SMEs. The consortium will make use of its existing relations to clinical collaborators in order to achieve proof-of-principle validation of the imaging modalities. The outcome contributes directly to improving and to maintaining the quality of life and living conditions of the European aging population through early diagnosis of cancer and of retinal pathologies as well as more efficient therapy monitoring. Moreover, the envisaged imaging modality may in the long term act as a screening device to investigate the prevalence of cancer as a function of geographic (regional) or gender related parameters. Finally, the diagnosis of other age-related diseases in a variety of medical fields, such as cardiology, neurology, gynaecology, and gastroenterology, benefit from this novel diagnostic platform provided by FUN OCT.
Agency: European Commission | Branch: H2020 | Program: SME-2 | Phase: SMEInst-05-2016-2017 | Award Amount: 2.48M | Year: 2016
1.3 million this is the sad number of Europeans dying each year by cancer. JenLab, high-tech company from Germany, has therefore developed an innovative novel diagnostic medical device (TRL 7) based on femtosecond laser radiation for immediate, non-invasive early diagnosis of cancer, particularly skin cancer, within seconds and with ultra-high resolution depicting even subcellular level. This is also true for early detection of ophthalmic diseases which constitute a major burden for Europes society knowing that one European in every 30 is expected to experience sight loss. In addition to intratissue cell imaging, JenLabs technology is able to measure metabolic processes giving information about diseases even before they become visible increasing therapy success by early detection and continuous monitoring. Moreover, the technology is applicable for imaging each tissue without labelling but using endogenous biomarkers. Outcome of the business innovation project is an miniaturised, certified and by clinical study validated ultracompact, flexible, fast multiphoton tomograph for dermatologists and ophthalmologists, representing an European market potential of EUR 11.4 bn. Particularly by its quick and reliable (using 4 different modes of diagnostic) way of diagnosis it meets user needs while simultaneously reducing Europes continuously growing health care costs. The business project is in line with JenLabs philosophy of saving lifes and of facilitate healing as well as it is in line with JenLabs strategy of developing and producing novel solutions for early diagnosis of life threatening and/or serious diseases.
Jenlab Gmbh | Date: 2012-10-05
A nonlinear laser scanning microscope for flexible, noninvasive three-dimensional detection comprising a measuring head which is flexibly connected to at least one radiation source by transmission optics and can be freely positioned in space, at least one controllable tilt mirror is arranged for aligning the excitation beam in order to keep the excitation beam concentric to an aperture-limited optical element of the measuring head, a test beam which is coupled out of the excitation beam onto a spatially resolving photodetector for monitoring the center alignment of the test beam as a conjugate position to the target position of the excitation beam and directional stabilizing the excitation beam by a control unit of the tilt mirror depending on a determined deviation.