Miscoscopy

Andor Dragonfly 200

Spinning disk confocal. Requires training.

Andor Dragonfly Spinning Disc Confocal Microscope

Training and self-service are offered.

The Andor Dragonfly 200 spinning disc confocal microscope provides high-speed, high-resolution imaging. Its configuration includes 10x and 20x dry, 40x water, and 63x glycerol objectives, alongside an additional 2x camera magnification, supporting subcellular level imaging. As a spinning disc system, the Dragonfly 200 allows for rapid image acquisition, making it an essential tool for real-time single-cell and molecular analyses. This instrument is able to accommodate live imaging of slides or wells.

Emission filters

Lasers

Excitation wavelengths are fixed: 405, 445, 488, 561, 637 and 730nm.

Keyence BZX-810

Keyence is not confocal, but it is pretty close to confocal and makes decent z-stacks using structured illumination.

The Keyence BZ-X800 is a versatile, all-in-one fluorescence microscope designed for high-resolution imaging and advanced analysis. It supports live imaging and accommodates multi-well plates, enabling efficient high-throughput studies. The microscope features four-color fluorescence, bright field imaging, tiling, and stitching for comprehensive sample coverage. It offers 3D views using structured illumination, allowing for precise z-stack imaging. This makes it ideal for detailed visualization of complex cellular structures and biological samples, and it is suitable for a wide range of research applications from basic cell biology to advanced biomedical studies.

High-resolution imaging
Live imaging capability
Multi-well plate compatibility
Four-color fluorescence imaging
Bright field imaging
Tiling and stitching for comprehensive sample coverage
3D views with structured illumination
Precise z-stack imaging

This 3D image was created using gCore's Keyence microscope. The z-stack imaging capability is enabled by a feature known as "Sectioning," which utilizes structured illumination microscopy (SIM) for 3D imaging. For training, please contact mrempel@ucsd.edu. The stained slide was provided by Gur-Cohen lab.

Keyence BZ-X810 Filter Cubes

49000-UF1 ET-DAPI (C211143): 358/461 nm (excitation/emission) - DAPI
49008-UF1 ET-mCH/TR (C211146): 560/630 nm - mCherry/Texas Red
49002-UF1 EGFP (C211144): 470/525 nm - Enhanced Green Fluorescent Protein (EGFP)
49006-UF1 ET-CY5 (C211147): 620/700 nm - Cyanine 5 (Cy5)

Objectives

4x PlanFluor - NA 0.13 - Working distance: 16.50mm
10x PlanApo - NA 0.45 - Working distance: 4.00mm
10x PlanFluor - NA 0.30 - Working distance: 14.50mm
20x PlanApo - NA 0.75 - Working distance: 0.60mm
20x PlanFluor - NA 0.45 - Working distance: 8.80-7.50mm
40x PlanApo - NA 0.95 - Working distance: 0.25-0.17mm
60x PlanApo - NA 1.40 - Working distance: 0.13mm

The long-distance objectives are in bold.

A higher Numerical Aperture (NA) gives more light and speeds up the imaging.

Imaris

gCore's Training videos

Imaris workstation is a commercial alternative to ImageJ, allowing automation of 3D reconstruction of cells, neurons, axons, dendrites, and vessels and also allowing automatic quantification of cells, nuclei, synapses, contact points, volumes and intensities.

We have a paid subscription to Imaris workstation, software and remote training. To schedule a remote training session, please write to Erich at ussupport_imaris@andor.com or at Imaris Remote Support Scheduler.

Included are IMARIS features:

gCore is subscribed to Imaris Single Full with ClearView (SFCV) license, which includes the following advanced image analysis tools:

Imaris Core: Allows rendering of 3D/4D images, along with interactive visualization, snapshot capture, and animation creation.
Imaris MeasurementPro: Facilitates the detection of 3D structures, enabling count, intensity, size, shape, and distance measurements, as well as object classification into user-defined classes, including options for machine learning.
Imaris Coloc: Supports pixel intensity-based colocalization analysis, including Mander’s, Pearson’s, and percentage overlap metrics.
Imaris Vantage: Provides tools to plot data in 1D to 4D and compare classes and groups using statistical tests.
Imaris XT: Offers custom analysis with integration for MATLAB, Python, Java, and FIJI. Users can utilize built-in scripts or XTensions made by the community, or create their own.
Imaris TrackLineage: Enables tracking of motion and morphology changes over time, detects divisions, and creates lineage trees.
Imaris Cell: Detects and analyzes cells (by membrane or cytoplasm) and organelles with per-cell statistics.
Imaris FilamentTracer: Assists in the detection, tracing, and analysis of filament-like structures, such as neurons and vessels.
Imaris Batch: Applies a saved protocol for batch analysis of multiple images.
ClearView Deconvolution: Utilizes GPU-accelerated deconvolution for enhanced image clarity.

These tools are included in our license, ensuring that you have access to a robust set of features for your imaging needs.

The following features are not included in our license:

Imaris Stitcher: Precise alignment and stitching of multiple image files.
Floating License Manager: Ability to share Imaris licenses between workstations.

Imaris features

Zeiss LSM780 confocal microscope

Located in Flow Core, rm 1301. Training is required.

The Zeiss LSM780 confocal microscope offers reliable imaging for cellular and molecular analysis. It features a GaAsP detector for improved signal-to-noise ratios and supports multiple laser lines (405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 594 nm, 633 nm) for flexible excitation of various fluorophores. The system allows multi-channel imaging and spectral imaging with a 32-channel GaAsP array detector, enabling the separation of overlapping spectra. It supports Z-stack acquisition for 3D reconstruction and time-lapse imaging for dynamic studies. Additional functionalities include high-speed scanning, photobleaching for FRAP experiments, and FRET analysis for protein interactions. The integrated Zen software provides tools for quantitative measurements and data visualization. The Zeiss LSM780 is suitable for diverse applications in cell biology and neurobiology, requiring standard fluorescence imaging capabilities.

Lasers

Olympus Confocal FV1000 FluoView

with Second Harmonic Generation and two-photon modes

The Olympus FV1000 FluoView microscope incorporates Second Harmonic Generation (SHG) and two-photon imaging modes, enabling advanced visualization of biological specimens with minimal disruption.

SHG mode facilitates label-free imaging of non-centrosymmetric structures, particularly collagen fibers. This non-invasive technique allows visualization of tissue architecture without exogenous stains, proving valuable for studying fibrosis and extracellular matrix remodeling. SHG imaging provides quantitative data on fiber orientation and density, offering insights into tissue structure and pathology.

Two-photon mode employs near-infrared light for deep tissue imaging, typically reaching depths up to 1 mm depending on tissue type. This technique offers reduced phototoxicity and photobleaching compared to single-photon excitation methods. It excels in applications such as in vivo brain imaging, tracking cell movements in intact organs, and long-term observation of living specimens. Two-photon imaging also provides enhanced three-dimensional resolution in thick, scattering tissues.

SHG - second harmonic generation, TPEF - two photon excited fluorescence by Olympus FV1000.

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