IVM Multi-Photon Facility

 The Intra-Vital Imaging (IVI) Multi-Photon Imaging Facility in the Houston Methodist Department of Systems Medicine & Bioengineering allows microscopic imaging of living tissue at the highest resolution possible with light microscopy. Olympus multi-photon excitation microscopy is an advanced fluorescence microscopy technique enabling subcellular resolution beyond the depth limits of conventional confocal microscopy. Using this technique, researchers can acquire high-resolution images within tissues of a living model. Furthermore, an additional SIM scanner and various associated laser beams enable simultaneous multi-photon imaging with direct laser stimulation. Researchers can use this system at Houston Methodist not only to view and capture remarkable 3-D images of tissues within living organisms, but also to stimulate and excite individual cells or networks of associated cells directly to probe biological mechanisms and develop therapeutic strategies.

In addition to the intra-vital microscope, the imaging area is equipped as a surgical suite containing a number of small instruments dedicated to physiological monitoring. Blood-gas analysis, electrolyte levels, blood pH, blood pressure and a number of other factors can be monitored both during surgical procedures and intra-vital imaging. This provides researchers at Houston Methodist with the ability to monitor and control the biological state of the subject visualized precisely, helping to ensure the validity of scientific results. The multi-photon instrumentation and associated physiological monitoring capabilities of the IVI imaging facility supports the translational research efforts of the Houston Methodist, that is, contributing to the unearthing of novel findings and strategies to lead to improved patient care.

Equipment Available at Intra-Vital Imaging (IVI) Multi-Photon Imaging Facility 

Full R6-416 equipment

FluoView FV1000 laser scanning microscope with SIM scanner and multi-photon imaging

  • Olympus BX61WI upright microscope
    • Maximum workspace allowing intra-vital imaging of small animals
  • SIM scanner with multi-combiner
    • Targeted laser stimulation while allowing simultaneous imaging
    • Two independent beam outputs with full control
    • Dual-port laser combiner enables lasers to be shared between stimulation and imaging
  • Mai Tai DeepSee Ti:Sapphire femtosecond laser
    • Optimized for multi-photon imaging
    • Tunable wavelengths from 690 to 1040 nm
    • 2.4 W of average power
  • Lasers: 405 LD, 459 Multi Line Argon, 488 Argon, 515 Multi Line Argon, 559 LD, 635 LD
  • Fluorescence illumination module
    • Mercury lamp with motorized shutter for epifluorescence imaging
  • Transmitted light detection module
    • Simultaneous multi-channel fluorescence confocal and transmitted DIC imaging
  • LSM, R690, VBYIR, GFP, CY3, DICT filter sets in Turret
  • Objectives
    • Piezo nano positioning objective scanner
    • PI-E665 LVPZT amplifier/servo controller
    • PI 721.CDQ PIFOC piezo flexure objective scanner
    • Sub-nanometer resolution with millisecond settling time
    • Travel range of 100 um
  • Siskiyou Model 100cr motorized X, Y stage with ASI controller
    • Automated with Fluoview software for multi-area imaging
    • 24 mm range of movement in both X and Y directions
  • OLY-150 Camera, controller with Rainbow CCTV
    • Real time imaging

Olympus SZX10 research stereomicroscope

    • High NA of 0.2
    • Mounted on an X, Y, Z movable stand for use during microsurgery

Shutter Instruments ROE-200 manipulator controller with two manipulator arms

    • Both arms are independently controlled by a single controller
    • Automated pipette exchange

Narishige IM300 microinjector

    • Programmable
    • Precise pneumatic control of small injection volumes

Harvard Apparatus homeothermic monitor

    • Feedback thermoregulation for intra-vital studieWarner TC-344B dual automatic temperature controller

Warner TC-344B dual automatic temperature controller
Grass Technologies S88 stimulator

    • Independent dual channel output stimulator
    • Single cell to muscle stimulation

Molecular Devices Axon CNS multiclamp 700B

    • Microelectrode amplifier for patch clamp, voltammetry/amperometry, ion-selective measurements or bi-layer recordings
    • Patch voltage-clamp or high-speed current clamp recording

Molecular Devices Axon CNS Digidata 1440A

    • 16-bit high-resolution data acquisition system
    • Digitizer with up to 8 simultaneous cell recordings
    • Acquire data through the accompanying AxoScope software

SAR-830/AP ventilator

    • Volume or pressure cycled operation for controlling respiration of small animals

Harvard Apparatus Inspira ASV 

    • Small animal respirator
    • Pressure monitoring, sigh breath, assist mode, and variable I:E ratio

VetEquip anesthetic instrumentation

    • Complete anesthetic setup with dedicated areas for animal prep, surgery, imaging, and recovery
    • Use in conjunction with small animal ventilators

Multi-gas setup

    • Allows mixing of various gases in any proportion desired
    • Create hypoxic, anoxic, or hypercarbic conditions
    • Use in conjunction with small animal ventilator, and/or anesthetic equipment

World Precision Instruments Pressure Monitor BP-1

    • Monitor animal arterial or venous blood pressure
    • Display Systolic, diastolic or average pressure

ACE Light source with dual arms

    • AC halogen light source

Omnidrill 35

    • High-torque tool for grinding, finishing and drilling bone

Germinator 500

    • Dry sterilize surgical Instruments in seconds

Stephen Wong, Ph.D, PE
R6-211, stwong@houstonmethodist.org