Multi-Photon Microscopy Core

Leica TCS SP5 AOBS Multi-photon Microscope

This dedicated intravital multiphoton imaging system is based on an inverted Leica DMI 6000 microscope powered by a fully automated, broadly tunable (680-1080 nm) Chameleon Ultra-II modelocked Ti: Sapphire MPE laser (Coherent). The system also has visible range Argon (458-514 nm) and He-Ne (543, 594 and 633 nm) lasers for conventional confocal microscopy. The six detector channels include two external, non-descanned high-sensitivity Hamamatsu PMTs for intravital multiphoton imaging, one transmitted light detector and three internal, pinhole detectors for conventional confocal imaging. A fully equipped small animal surgery room is directly adjacent, and used for preparation of the animal for intravital imaging. Additional instrumentation around the microscope includes heat-isolation stage plate and body temperature control via thermal blanket (Harvard Apparatus), continuous iv fluid minipump and mobile blood pressure measuring system (BP-1, WPI) for simultaneous maintenance and recording of cardiovascular parameters during intravital imaging, and rodent anesthesia machine (E-Z Systems) with nosepiece for continuous isoflurane anesthesia of mice and rats.

The mandate of this core facility is to provide access to multi-photon in vivo imaging of intact organs in small animals and this state-of-the-art technology is recognized for its capability of deep optical sectioning of living organs. It enables USC investigators to perform ultra-sensitive, quantitative imaging of organ functions in health and disease with high spatial (submicron) and temporal (50-1000Hz) resolution that other imaging modalities cannot achieve. The core has extensive experience in intravital multi-photon imaging of various organs in the rat and mouse, including the skin, kidney, spleen, liver, pancreas (both exocrine and endocrine), intestine, and bone. Recent applications include serial, intravital multi-photon microscopy of the same intact tissue area/volume over days for tracking the fate of single somatic or stem cells in the living mouse kidney (Hackl et al Nat Med 2013) and detection of changes in cell calcium in vivo in the intact kidney during a disease process (Burford et al J Clin Invest 2014).

The usage charge for the SP5 multi-photon system is $50/hour.

Leica SP8 DIVE Multiphoton Confocal Microscope

This new model, the Leica SP8 DIVE MP system is an advanced microscope to increase tissue depth penetration for intravital imaging of highly scattering intact organs and tissues. This is achieved by the combination of at least three features of the SP8 DIVE system: long wavelength excitation using extended IR spectrum multiphoton laser (680-1300 nm with the Discovery laser system from Coherent), ultra-high speed resonant scanning (8 kHz), and ultra-high sensitivity quad module four spectral HyD SP GaAsP detectors (first in class). The Leica SP8 DIVE system comes with an DMi8CEL advanced modular inverted microscope, one high sensitivity Hamamatsu PMT and two internal HyD detector channels, AOBS scan head, RYB confocal laser kit for AOBS, long working distance (up to 2.4 mm) 25X, 40X, and 63X MP objectives, motorized scanning stage, LAS X SP8 and 3D visualization software and Huygens package.

The usage charge for the SP8 DIVE confocal system is $50/hour.

The Core is available to the entire USC community by appointment. For appointment and initial training, please contact the core director or manager directly.

Watch Videos

• Pod-Confetti movie showing z-sectioning of an intact renal glomerulus in vivo in which podocytes were labeled in multi-color by the confetti reporter.
• Pod-GCaMP3 movie showing cell-to-cell propagating calcium wave in vivo

Contact Info:

Director Dr. Janos Peti-Peterdi,
Professor of Physiology and Biophysics, and Medicine
Office: (323) 442-4337
Email: petipete@usc.edu

Location

Health Sciences Campus
Zilkha Neurogenetic Institute
Room ZNI 332
1501 San Pablo Street,
Los Angeles, CA 90033