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RatioMaster™
Fluorescence Microscopy

 
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PTI's time tested RatioMaster™, offering researchers solid, dependable, sensitive detection for the collection and analysis of ratio-metric photometry data for calcium, pH, and intracellular ion imaging!

The RatioMaster™ is a microscope-based ratio spectrofluorometer capable of dynamic ratio fluorescence measurements on a millisecond timescale. A xenon arc lamp provides high intensity, continuous broadband illumination. Alternating excitation wavelengths are selected by a computer-controlled high-speed random access monochromator coupled to a (user specified/supplied) inverted fluorescence microscope with a liquid light guide. Emitted light is collected from the sample and passed through a photometer having a bilateral translatable iris and a viewing eyepiece to a switchable analog/photon counting photomultiplier detector. Analog detection is used when emitted light is relatively intense; low light levels are detected by photon counting. All system functions are under computer control. Data is collected and analyzed by proprietary Windows based advanced fluorescence software.

PTI RatioMaster System

What's in the Box

  • DeltaRAM X™ monochromator
  • 2 meter liquid light guide
  • D104 microscope photometer
  • BryteBox acquisition hardware
  • Software
  • Installation and training

General Applications and Dyes

Excitation-Shifted

Application

Dye

Calcium Fura-2,
Fura PE3
FFP18 Quin-2
Fura-Red
Sodium SBFI
BCECF
HPTS
SNAFL®
DM-NERF®
CI-NERF®
Potassium PBFI
Magnesium Mag-Fura-2
Mag-Fura-5
Membrane Potential Di-4-ANEPPS
Di-8-ANEPPS

Emission-Shifted

Application

Dye

Calcium Indo-1
pH SNARF-1®
SNARF-2®
Magnesium Mag-Indo-1
Membrane Fluidity Pyrene Fas
PATMAN
Free Fatty Acids ADIFABTM
Membrane Potential JC-1
Lipids, phoholipids Nile Red
Nucleic acids Acridine
Orange

Calcium-related Measurement

  • Ca2+ transients
  • Ca2+ mobilization
  • Ca2+ homeostasis
  • Ca2+ waves
  • Ca2+ oscillation
  • Ca2+ spikes
  • Ca2+ entry
  • Capacitative Calcium Entry (CCE)

Calcium Measurement

  • Intracellular free Ca2+
  • Mitochondrial Ca2+
  • Endoplasmic reticulum (ER) Ca2+ pool
  • Lysosomal Ca2+
  • Extracellular near membrane Ca2+

Simultaneous Measurement of intracellular a2+ and:

  • Cell volume
  • Cell contraction: Edge detection device
  • Membrane current: Patch Clamps
  • Phygocytosis
  • Cl-
  • Oxidase activation
  • Ca2+-CaM

Membrane Potential

  • Plasma membrane Potential
  • Mitochondrial Membrane Potential

pH

  • Intracellular pH
  • Phagosomal pH
  • Vacuolar pH
  • Lysosomal pH
  • More..

Non-Ratiometric

Application

Dye

Calcium Fluo-3
Calcium Green™
Calcium Orange™
Calcium Crimson™
Rhod-2
Chloride SPQ SPA
MQAE
HIV protease HIV protein substrate
F-actin phallotoxins
PKC Fim-1
Rim-1

Dual Dye

Application

Dye

Nucleic acids DAPI and FITC
FITC and APC
Fura Red™ and BCECF
FITC and Texas Red®
DAPI and PE

FRET

  • Measure molecular proximity
  • Binding of enzyme to its substrate
  • Measure Endoplasmic reticulum (ER) Ca2+
  • Donor Excitation Spectroscopy
  • Emission intensity
  • Donor emission intensity
  • Acceptor emission intensity
  • Ratio of donor/acceptor emission intensity

Common FRET Pairs

  • EGFP / DSRed2 FRET
  • CFP / DsRed2 FRET
  • BFP / DsRed2 FRET
  • CoralHue M-Cyan / K-Orange
  • GFP / Rhod2
  • FITC / CY3 FRET
  • FITC / TRITC FRET
  • YFP / TRITC FRET
  • CY3 / CY5 FRET
  • CFP / YEP FRET
  • BFP / GFP FRET

Other Ions

  • Intracellular Mg2+
  • Intracellular Mn2+
  • Intracellular Zn2+
  • Intracellular Sr2+
  • Intracellular Na+
  • Intracellular Cl-
  • Intracellular Ba2+
  • Divalent cation influx
  • Labile iron pool (LIP)
  • Iron homeostasis
  • Na+ / H+ exchange
  • H+ / HCO3- exchange
  • Pf and PCO

Other Measurements

  • Respiratory burst & reactive oxygen species (ROS)
  • NAD(P)H
  • Nitric Oxide (NO)
  • Intracellular GSH
  • Identifying cells coexpressing GFP for analysis
  • Use ratiometric GFP (redox-GFP) to measure cellular/mitochondria reduce/oxidize reaction
  • Calcein-based cell viability assay
  • Oxygen consumption in cultured cells
  • And Many More..

 

 
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