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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.
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..
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Non-Ratiometric
Application |
Dye |
| Calcium |
Fluo-3
Calcium Green™
Calcium Orange™
Calcium Crimson™
Rhod-2
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| 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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