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RatioMaster™ SL200
Fluorescence Microscopy

 
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Applications

RatioMaster™ is up for the task for many applications
RatioMaster

[Ca2+] signals in cardiac myoblasts and myotubes

More Applications

  • IP3-induced intracellular
  • [Ca2+]c and mitochondrial [Ca2+]m
  • IP3-induced intracellular [Ca2+]c and mitochondrial[Ca2+]m
  • Visualization of Mitochondria by mitoGFP
  • [Ca2+]m signals in permeabilized myoblasts and myotubes
  • IP3-induced intracellular [Ca2+]c and mitochondrial [Ca2+]m

 

In-Situ Calibration of Intracellular [Ca2+]i

RatioMaster

By measuring the ratio of fluorescent intensity at 340/380 nm, [Ca2+]i can be measured over several orders of magnitude and with a high degree of precision.

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Simultaneous measurement of Ca2+ and myocyte cell length with Felix 32

RatioMaster

The above figure shows data traces from the simultaneous collection of fluorescence and cell length from a cardiac myocyte. Myocyte was loaded with Indo-1 and excited at 365 nm wavelength using the RatioMasterT RM-5 dual emission detection system. The blue trace shows the calcium ratio increase and decrease with the cell contraction . The contraction data (Green trace) can be correlated with sample accuracy with the fluorescence data since both signals were collect simultaneously.

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FRET for molecular proximity assay

FRET occurs because the seprase and the plasminogen activator receptor co-localize on the membrane of malignant melanoma cells. FRET was detected via a fluorescently labeled antibody targeted to each receptor. The results of this FRET: The acceptor's excitation spectrum gains spectral features of the donor's excitation spectrum. Not a real FRET, but similar. Instrument: PTI RM3.

RatioMaster

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FRET for Binding Assay

RatioMaster

FRET occurs when the EYFP fusing RII protein kinase binds to the ECFP fusing Ht31 protein (kinase anchoring protein). The results of this FRET: The donor (ECFP) emission fluorescence decreases. The acceptor (EYFP) emission fluorescence increases. Instrument: PTI's FRET RM pro.

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FRET to measure Ca2+ in ER

FRET occurs when ER Ca2+ binds to the "chameleon 3er", causing it to fold thereby bringing the two dyes in close proximity. The results of the FRET: The donor (CFP) emission fluorescence decreases. The acceptor (YFP) emission fluorescence increases. The emission ratio of Acceptor/Donor increases. InsP3 triggers the release of ER Ca2+, thus decrease the level of FRET. Instrument: PTI's FRET RM pro.

RatioMaster

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