Microscopy and Image Analysis Core (MIAC)

MDRC investigators receive consultation prior or during training

Advice focuses on immunological staining of fixed cells/tissues, fluorescent-based dyes for live cell imaging, microscopy techniques, processing and visualization of microscopic images, and morphometric features analysis

Core Director and Laboratory Director typically handle the consultations

Recommendations to other experts from the UM community provided when needed

Investigators can use Core equipment at an hourly rate or obtain services by core personnel for additional fees

The core has a Nikon A1, a Leica Stellaris 8, and a Leica TCS SP5 Confocal Microscope

Each system has its strengths to optimize acquisition for a variety of imaging configurations

Investigators are guided to the confocal system that fits best their needs

Images can be analyzed by users after training or Core personnel

Users receive four free hours of instruction

The service includes morphometric image analysis and preparation of publication-quality images

The Core has software programs like AutoQuant and Leica's Lightning for image restoration

Quantitative analysis of images can be done using MetaMorph, ImageJ, Aivia or MATLAB software

3D reconstruction and fluorescence intensity quantification can be performed with Imaris and Aivia software

Publication-quality figures and montages preparation available with Photoshop, Illustrator, Prism, and Powerpoint software

Instruction in the use of software provided by Laboratory Director

An inverted Nikon TiE microscope is available for imaging and standard fluorescent signal acquisition (blue, green red and far-red) with a sensitive Photometrics CMOS camera

Equipped with specialized excitation and emission filters to allow users to capture fluorescence-based images of transfected green fluorescent protein (GFP) chimeric molecules, eCFP and eYFP signals for Förster Resonance Energy Transfer (FRET)

Image acquisition are done with Nikon Elements software

Consultation begins with the Laboratory Director to decide appropriate microscopy technique between confocal and wide-field fluorescence microscopy

Stage-top devices available for temperature and CO₂ regulation

Support a variety of dyes and probes for sensing, labeling, and tracking 

The core has experience to date using fura-2, fluo3, calcium green and Magfura dyes for Ca²⁺, SBFI for Na⁺, BCECF for H⁺, SPQ for Cl^-, cSNARF-1 AM for pH, NADH/NAD+ sensors, lucifer yellow and lissarhodamine as gap junction tracers, and eCFP, eGFP, eYFP, DSRed, mCherry fluorescent proteins as markers for transfected cells, membrane potential sensors (TMRM, TMRE, JC-1), labeling mitochondria (MitroTracker and Bacmam MitoGFP/RFP) and following endocytosis

Frequency of image collection can be optimized to manage data files size and minimize probe bleaching

Consultation and training available for complex imaging techniques like FRET, FRAP, FLIP, FLIM, photoactivation and uncaging 

Nikon A1 and Leica Stellaris 8 systems have specialized protocols for image acquisition

Instrumentation and training for FRET is available but FRET is not offered as a routine service since it requires a large developmental component