Smart imaging devices for Bio Imaging Lars Hufnagel
Smart imaging devices for Bio. Imaging Lars Hufnagel EMBL Heidelberg In collaboration with Hamamatsu Photonics and Luxendo Gmb. H
Deciphering molecular basis of life requires cross scale imaging > Imaging technologies that can cross spatial and temporal scales <1 um <100 um Systematic map of molecular mechanisms at the cellular level > 1 mm Mechanistic understanding at tissues, organisms and disease New CRISPR genomic engineering tools allow to specifically label and manipulate almost every gene/protein in living cells.
Fast 3 D imaging Light-sheet microscope 200μm Confocal microscope area detector point detector 800 seconds ~number of planes In toto embryo imaging time Photo-toxicity <1 seconds light sheet thickness Pioneered at EMBL Heidelberg Huisken, et al, 2004
Imaging biological specimen across scales 3 D live imaging Zebrafish brain Fruit fly Krzic et al, Nat Methods 2012 Light sheet nanoscopy Mouse embryo Strnad et al, Nat Methods 2015 Quantitative measurements Capoulade et al, Nature Biotechnology 2011 P. Hoyer, J. Engelhardt, S. Hell, L. Hufnagel PNAS 2016
Where will we go? feasible today 2 Mpixels 5 TB/day 4 Mpixels future 8 -16 Mpixels >50 Mpixels 23 TB/day >160 TB/day
Realtime data processing Phallusia imaging Digital representation GPU and FPGA based processing GPU–based realtime compression library for fluorescent images with up to 1 GB/s yields 1/30 compression Realtime data processing and segmentation
On sensor scattering discrimination dumped constant reset readout activation front Gustavo de Medeiros, et al Nat Comm 2015 Patent Pending
Summary • Light sheet microscopy allows 3 D imaging of biological specimen at low light levels for extended periods of time. • Light sheet microscopy is compatible with a vast variety of molecular imaging techniques. • Integration of realtime data processing will be vital for future applications Light sheet imaging would profit from: 1. Gated or time-resolved imaging (~ns) • molecular environment • protein-protein interaction • kinetics, concentrations 2. Large (>16 MP) small pixel size (1 -2 um) sensor with low noise (<1 e rms) with high sensitivity (QE>80%) in VIS to NIR range. 3. Integration of data processing and imaging 4. Hyperspectral imaging in the VIS range
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