Live Imaging and Other Capabilities of the Bioscience Imaging Facility (BIF)

The BIF provides the instruments and expertise needed to visualize molecules in preparations ranging from single cells to entire animals. Project specific instrument training & advice Consultation on sample preparation, image processing, and data analysis Charge-back rates; Collaborations are encouraged The Bioscience Imaging Facility

High NA oil objectives 10x, 20x, 40x, 60x Seven visible laser lines 446, 458, 488, 514, 561, 640 nm Four channels or 32-bin spectral detection Resonant scanner* Piezo stage* Live imaging chamber* *Ideal for live imaging monolayer cell cultures Rotate and Adj High NA oil objectives 10x, 20x, 40x, 60x Seven visible laser lines 446, 458, 488, 514, 561, 640 nm Four channel detection High NA oil objectives 10x, 20x, 40x, 60x Seven visible laser lines 446, 458, 488, 514, 561, 640 nm Four channels or 32-bin spectral detection Resonant scanner & Piezo stage* Live imaging chamber* *Ideal for live imaging monolayer cell cultures High NA oil objectives 10x, 20x, 40x, 60x Seven visible laser lines 405, 458, 488, 514, 561, 640 nm Four channel detection Nikon A1R Nikon C1+ Confocal Microscopy

Resonant Galvanometer Piezoelectric z-stage High Speed Image Acquisition

Spectral Acquisition --Document spectra --Remove autofluorescence --Spectral FRET --Create virtual channels T(λ) = C 1 R 1 (λ) + C 2 R 2 (λ) Other Uses: Spectral Unmixing Fluorophores in sample Spectral Detector

LWD (500 um) water 40x objective* Mai Tai DeepSee tunable IR laser* Four channel non-descanned detection* Resonant scanner* Live imaging chamber* Confocal option Two laser lines (405, 488); four channel detection. *Ideal for live imaging deep into living tissues Nikon A1R_MP Multiphoton Microscopy

Phase and PlanFluor objectives* DS-Ri1 color camera (standard) Photometrics QuantEM CCD (optional)* Live imaging chamber* *Ideal for high sensitivity or high frame rate imaging of monolayer cell cultures Nikon Ti-S Wide-field Microscopy

Microscopic Images Juan Martinez, Mark Hall Biochemistry Pablo Zavattieri Civil Engineering Shawn Liu, Xiaoqi Lin Biology Aaron Taylor Bindley

Cooled CCD Range of excitation and emission filters Spans the nm Isofluorane anesthesia provided *Ideal for collecting signal from within an organism IVIS Lumina II In vivo Optical Imaging

In vivo Optical Images Cansu Cimen, Tim Ratliff Comparative Pathobiology Dan Szymanski Agronomy

Combined SPECT and x-ray CT Enables SPECT and CT image registration Multi-pinhole collimators High resolution imaging (350 um voxels) High sensitivity imaging (kBeq source) Data binning relative to biological rhythms Reduces blur List mode acquisition and statistical 3-D image reconstruction algorithm. Improves image quality MiLabs U-SPECT-II/CT Micro SPECT / CT Imaging

Principles of microSPECT

Principles of Statistical Reconstruction ? Object Space Projection Space

Micro SPECT / CT Images Xin Lu, Shuang Liu Health Sciences Charity Wayua, Phil Low Chemistry

Live Imaging Accessories

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