Confocal & Multiphoton Microscopy Unit
Anna-Liisa Nieminen, PhD
Associate Professor, Department of Drug Discovery & Biomedical Sciences
Location: Drug Discovery Building 507
The Confocal & Multiphoton Microscopy Unit of the Cell & Molecular Imaging Shared Resource provides:
- Live cell imaging of parameter-sensitive fluorophores to monitor ions, electrical potentials, oxygen, and nitrogen radical generation, NAD(P)H, mitochondrial and plasmalemmal membrane permeability, cell viability (apoptosis and necrosis), fluorescent protein biosensors, and other parameters
- High resolution imaging of tissue sections for immunocytochemistry and fluorescent protein distribution
- Fluorescence resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP) to characterize and quantify interactions between specific molecules and their mobility
- Intravital microscopy to monitor microcirculation, leukocyte margination, invadopodia, mitochondrial polarization, and permeability, radical generation, gene expression, detection of collagen fibers, and other factors in living animals and tissues
The Confocal & Multiphoton Microscopy Unit is equipped with the following instruments:
- Zeiss LSM 510 NLO META multi-photon laser scanning confocal/multiphoton microscope: Multiple lasers for excitation at 458, 477, 488, 514, 543, and 633 nm and multi-tracking permit simultaneous recording of multicolor fluorescence in the confocal mode. A recently upgraded Coherent Chameleon Ultra II Verdi laser provides multiphoton excitation tunable from 680 nm to 1080 nm. A META photomultiplier array detector provides spectral detection and unmixing for both 1-photon and multiphoton fluorescence. An environmental chamber controls stage and objective temperature and specimen atmosphere. Note, an NIH instrumentation application is pending that would support the upgrade of this instrument to a Zeiss LSM 780 34 Channel Quasar.
- Zeiss LSM 510 META: Except for multiphoton capability, this microscope has all the capabilities of the LSM 510 NLO META microscope described above.
- Olympus FluoView FV10i LIV: The Fluoview FV10i is a self-contained compact inverted confocal laser scanning microscope with water immersion optics and environmental control for live cell imaging. Four diode lasers provide excitation at 405, 473, 559, and 635 nm. Detection uses a variable bandpass filter allowing selection of the most suitable wavelength for a particular fluorophore.
- Olympus FV1200 MPE: The FV1200 MPE is a dedicated multiphoton microscope configured for intravital imaging of live animals and is scheduled for installation in the near future. This inverted microscope is equipped with long working distance water and silicone oil immersion objectives, tunable multiphoton laser with prechriping, and highly efficient GaAsP detectors for intravital multiphoton projects in the future and will uniquely evaluate tumor metabolism and cell biology in vivo in response to genetic manipulations, pharmacologic agents, and cancer chemotherapy drugs.
- Leica TCS SP2 AOBS: This upright laser scanning confocal microscope features a full complement of DIC objectives, dipping lenses, programmable acousto-optical beam splitter, and prism spectrophotometer for emission detection. Leica software includes capabilities for multicolor analysis, 3D reconstruction, quantification tools, spectral unmixing, FRAP, and FRET.
- Fluorescence Plate Reader: A BMG NOVOstar fluorescence plate reader can be used for parallel measurements of fluorescence indicators in cultured cells grown on multi-well plates.
The iLabs Reservation Software enables online scheduling for users of the Confocal & Multiphoton Microscopy Unit: https://my.ilabsolutions.com/account/login.
|Microscope||Base Rate/Hr||HCC Rate/Hr||After Hours Rate/Hr|
|Zeiss 510 (HCC Building)||$40||$30||$20|
|Leica (HCC Building)||$20||$15||$10|
|Zeiss 510 (Drug Discovery)||$40||$30||$20|
|Zeiss 510 (Drug Discovery, Multiphoton Laser||$48||$36||$24|
|Olympus 10i LIV (Drug Discovery)||$40||$30||$20|
|Assistance/Training by Staff||$50||$37.50||Same|