State-of-the-art fluorescence imaging of organized biochemical activities at the molecular, cellular and tissue levels.

High Content Microscopy

This instrument is equipped with objectives ranging from 1x to 100x, 0.05 to 0.95 NA for air and 1.4 NA for oil, capturing a variety of cellular and intra-cellular events and is capable of capturing > 10 million cells/hour in a low-resolution, whole-well, 3 color cell scoring application, or > 1 million cells/hour in a high resolution two-color assay for either compound or genetic screening applications.  A robotic fluidics handler allows kinetic manipulation of cell signaling pathways via the interactive addition of compounds.

Location: SDSU
Contact: Adam Hoppe

Multiphoton Microscope

The multiphoton microscope is outfitted with a 25X water dipping objective with a numerical aperture of 1.05, 2 mm working distance, and correction collar capable of capturing bright images from deep focal points with a relatively wide field of view.  The high scanning speed of this instrument enables capture of dynamic real time intravital imaging data in living organisms.  Frame rates of 30 fps can be achieved in 512 x 512 pixel fields with a maximum of 438 fps possible in narrower fields of 512 x 32 pixels.  This imaging system is equipped with two infrared wavelength femtosecond pulse lasers (InSight 680-1300 nm, MaiTai 690-1040 nm) enabling high powered multicolor imaging with minimal phototoxicity, photobleaching, or tissue damage.  The additional laser light stimulation SIM scanner supports simultaneous activation and read-out imaging from techniques such as uncaging, photoactivation, or FRAP.

Location: SDSU
Contact: Darci Fink, Sen Subramanian

Multiphoton

Median optical section of a soybean root tip (left) and an emerging lateral root (right) expressing fluorescent sensor constructs to localize outputs from two major hormones auxin (green) and cytokinin (red).

Atomic Force Microscopy (AFM) and Photo-Activated Localization Microscopy (PALM)

Capable of resolutions nearing 10 nm in three dimensions, PALM and its variations provide unprecedented access to the subcellular organization of molecular complexes. Dr. Steve Smith has recently assembled and tested a PALM imaging system at SDSMT. Dr. Smith and new faculty hires will work to develop new infrastructure for integrated PALM & AFM and other techniques for nanoscale microscopy in the BioSNTR

Location: SDSMT
Contact: Steve Smith

AFM

(a) Composite fluorescence image of an unroofed SKMEL cell, composing of cyan fluorescence from Tq2-tagged clathrins and green fluorescence from F-actin filaments immunolabeled with green phalloidin antibodies; (b) composite AFM image comprising of eight individual 10 µm ×10 µm AFM images, and (c) overlaid of fluorescence-AFM images.

High-Resolution Transmission Electron Microscopy (HR-TEM)

To complement the superresolution fluorescence methods proposed here, the High-Resolution Transmission Electron Microscopy (HR-TEM) laboratory at SDSMT is equipped to accommodate biological samples. This facility is managed by Dr. S. P. Ahrenkiel, who has decades of experience in HR-TEM, including experience in cryo-micro sectioning of biological samples, tomographic imaging and 3D image reconstruction.

Location: SDSMT
Contact: Phil Ahrenkiel

HR-TEM

 

Lattice Light Sheet Microscope

The lattice light-sheet microscope is a refinement on the idea of selective-plane illumination microscopy (SPIM), where sample excitation is a thin sheet of light, sectioning the sample and reducing background fluorescence, photo-bleaching, and photo-toxicity.  Sampling frame rates are dramatically improved by widefield imaging of the illuminated plane with the detection objective oriented normal to the excitation plane.This instrument will enable acquisition of spatiotemporal information in living cells not possible on any existing microscope systems and will play an important role in accessing information about molecular organization and dynamics.

Location: SDSMT
Contact: Robert Anderson

Lattice Light Sheet

(A) Lattice light-sheet microscope with control rack (background), (B) LLSM without enclosure, (C) laser combiner without enclosure.