Quantitative microscopy assays - Fluorescence, bright field, confocal and two-photon

Image Analyst MKII supports a range of fluorescence microscopic imaging assays by generic protocols for data recording on typical microscopy systems and specific analysis protocols using pre-configured pipelines accessible from the main menu.

• • All quantitative microscopy

  • Intensity-based

  • Morphology-based

  • Cell Physiology

  • Mitochondria

  • Ca²⁺

  • ROS

  • With protocol

  • With video

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  Intracellular ion measurements with ratiometric dyes (Fura-2, SBFI, PBFI) (2,9,16,18,19,25)
  Video

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  Intracellular calcium measurements with non-ratiometric dyes   (Fluo-4, Rhod-2, X-Rhod dyes)(1,3,18,28)
  Pipeline

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  Mitochondrial calcium measurement using the high pass filtering technique (1,3,18,43)

• 

  Mitochondrial NADH/NAD⁺ measurement using the high pass filtering technique (29)
  Protocol Tutorial data

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  Mitochondrial or plasma membrane potential measurements (3,7,8,16,19)
  Pipeline

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  Absolute calibrated of mitochondrial and plasma membrane potential measurements (10,14,17,26,30,31) Protocol Video More

• 

  FRET calculations (9)
  Pipeline Tutorial data

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  Fluorescence bleed-through correction (9,16)
  Protocol Tutorial data

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  Segmentation-based, automated analysis of fluorescence intensities (8,13,16,19,25,26)
  Protocol Pipeline More

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  Reactive oxygen species measurements (26) Protocol DCFDA Protocol DHE Protocol MitoSOX Video Tutorial data

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  Mitochondrial swelling assay using optimized spatial filtering (6,11,22,30)
  Protocol Tutorial data

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  Organelle motion assay using optical flow (7,38)
  Protocol Tutorial data

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  Segmentation based morphological assays, adaptive thresholding (8,10,13,16,19,25,26)
  Pipeline 1 Pipeline 2 More

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  Quantification of intramitochondrial phosphate precipitates in electron microscopic images (10)

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  Analysis and quality control of synaptosome preparations (8,13,19,25). Selective imaging of dopaminergic nerve termini by image segmentation and gating (13)

• 

  Measurement of mitochondria to cell volume fraction with confocal microscopy (13,14,17,21,24,26,31)
  Protocol Tutorial data

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  Counting protein aggregates in c. elegans (15)

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  Whole-well cell counting for normalization of Seahorse respirometry data (12)
  Video

• 

  Cell tracking at low temporal resolution, for microplate imaging (unpublished)
  Pipeline

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  Senescence associated β-galactosidase (SA-BGAL) densitometry (unpublished)
  Pipeline

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  Apoptosis vs. necrosis cell count (unpublished)
  Pipeline

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  Assay of succinate dehydrogenase activity in cell culture by measuring formazan absorbance normalized to SDHB immunofluorescence (32)
  Pipeline - coming soon...