Analytic Microscopy

Purpose

The Analytic Microscopy Core’s (AMC) purpose is to provide access to the latest optical microscopy imaging technologies and expertise. The Core provides training and assisted support for many of its microscopes and imaging analysis software. Core staff members are available to assist investigators in the experimental and scientific design of their imaging experiments so that the correct reagents, staining protocols and imaging techniques are employed to best address the scientific questions of the investigator. The AMC staff works closely with members to design, image, and analyze experiments, allowing members to obtain high-quality and reproducible data. Further, the AMC collaborates with other Shared Resource Facilities at MCC to manage complex projects and to ensure proper handoffs of materials and data.

Services

Opera Phenix High-Content Screening

• Confocal imaging for live cell experiments or endpoint assays
• 4 laser/detector channels for DAPI/FITC/TRIC and Cy5 (or similar fluorophores)
• 20x, 40x and 60x High-resolution water immersion objectives
• Image multi-well plates, slides and even 3D-printed plates with unique formatting
• Robotic abilities to move plates from an attached incubator to instrument
• Liquid handling enables the automatic addition of reagents or compounds during the experiment
• Manages up to 80 plates at one time
• Includes Harmony software for full HCS experimental design, setup and analysis

Leica SP5 AOBS Multiphoton Confocal Microscope:

• 9 tunable laser lines and MP Ti-sapphire laser
• 4 PMT and 2HyD detectors
• 3D imaging and reconstruction
• Colocalization assays
• Multiphoton imaging
• FRET & FRAP
• Calcium imaging
• Second Harmonic Generation
• Environmental chamber for animal and live cell imaging

Leica SP8 Confocal Microscope:

• 4 laser lines and 2 PMT detectors
• 2 HyD detectors
• Colocalization assays
• 3D imaging and reconstruction
• An incubation/hypoxia chamber is available for live cell imaging
• Fitted with a 40x, 63x and 100x oil objectives with additional zoom software function enabled

Widefield Fluorescence Microscopes:

• Fully automated upright and inverted Zeiss microscopes
• LED light source and detection capabilities of 350-850nm
• Tile scan and z-stack acquisition with post-image processing
• Deconvolution
• Fully enclosed incubation chamber for long-term time-lapse live cell imaging
• Apotome available for pseudo-confocal imaging
• EVOS FL simple observation and acquisition
• EVOS Auto including tile scan and live cell imaging

PhenoCycler-Fusion

• Akoya Biosciences HT system
• Whole slide multi-plex staining (performed in-house) with barcode technology allows this system to capture 100+ biomarkers
• Accommodates FFPE, Fresh frozen or TMA samples
• Images fluorophores ranging from AF488-AF750 (plus DAPI) and optimized for Opal fluorophores

Intravital Microscopy:

• Live animal imaging via dorsal skin flap or cranial window chambers
• Analyze tumor growth, invasion, collagen or vasculature modeling longitudinally
• Real-time analysis of angiogenesis, blood flow, vascular permeability, drug/agent kinetics
• Real-time microscopy quantification of pH or other metabolomic gradients

Aperio ScanScope AT2 Slide Scanner:

• Holds up to 400 slides; scan time 30 slides/hour
• Segments TMAs into individual cores
• Eslide manager software capable of positive pixel, nuclear, cell, membrane, and vessel algorithms
• Genie software capable of segmenting regions of interest for IHC analysis

Incucyte S3 Two-Color FL  and SX5 Three-Color FL systems:

• High throughput live cell microscope with phase contrast and green/red FL – plus far red FL in SX5
• 96-well Wound Maker for wound healing assays
• Spheroid, migration, metabolism and Chemotaxis experimental modules
• Various assay kits are available through Sartorius

Image Analysis:

• Definiens Developer
• Definiens Tissue Studio
• Visiopharm
• Imaris 3-D
• Media Cybernetics Image Pro Plus
• Leica and Zeiss software

Also available:

• Laser Capture Microdissection
• Stereoscopy

Location

The AMC is located in the Moffitt Research Center (MRC) on the fourth floor, room 4058. An additional satellite facility is located in the Stabile Research Building (SRB) on the ground floor, room 20307. This facility houses the Leica SP5 AOBS Multiphoton Confocal Microscope.

Scheduling

The AMC uses a Laboratory Information Management Suite (LIMS) located on the Moffittnet for all Core requests, scheduling, billing and usage tracking.

Publications

All publications based on work conducted in the Analytic Microscopy Core should acknowledge the facility. A suggested statement is as follows:

If a Core Staff member(s) significantly contributes or participates in a project, investigators are encouraged to consider including the individual(s) in the list of authors.

Contacts

Scientific Director

Marilyn Bui, MD, PhD
marilyn.bui@moffitt.org

Core Facility Manager   

Joseph Johnson
joseph.johnson@moffitt.org

Office: 813-745-8710
Lab Number: 813-745-6157     

E-mail for all Core staff and incucyte/Opera Phenix reservations:
AMC@moffitt.org

Download Brochure (PDF)