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 | Detection and Modulation of Cariomyocyte Calcium Oscillations Using the FDSS6000- Author: Hamamatsu Photonics, K.K.
- Source: Hamamatsu Photonics, K.K.
- Date: Aug-06
- Abstract: Current methodology to detect calcium oscillations in mouse fetal cardiomyocytes uses plated on cover slips or Petri dishes. Using this model cells are exposed to putative cardiomodulators to investigate their pharmacological We report a methodology to transfer this model to high throughput screening using the FDSS6000. Wells show a baseline calcium oscillation (pulse) rate of either about 1/sec or appear quiescent. In both well types the agonist Isoproterenol increased pulse rate. In the latter wells we show the antagonist Propranolol blocks Isoproterenol agonism.
| | Detection and Modulation of Cariomyocyte Calcium Oscillations Using the FDSS6000- Author: Hamamatsu Photonics, K.K.
- Source: Hamamatsu Photonics, K.K.
- Date: Aug-06
- Abstract: Current methodology to detect calcium oscillations in mouse fetal cardiomyocytes uses plated on cover slips or Petri dishes. Using this model cells are exposed to putative cardiomodulators to investigate their pharmacological We report a methodology to transfer this model to high throughput screening using the FDSS6000. Wells show a baseline calcium oscillation (pulse) rate of either about 1/sec or appear quiescent. In both well types the agonist Isoproterenol increased pulse rate. In the latter wells we show the antagonist Propranolol blocks Isoproterenol agonism.
|  | Effect of Plate Mixing, Fluid Addition Height and Speed on Reducing Addition Artifacts and Negative Control Drift Using the FDSS6000- Author: Hamamatsu Photonics, K.K.
- Source: Hamamatsu Photonics, K.K.
- Date: May-06
- Abstract: Two artifacts often present following fluidic addition to wells are: 1) Sharp change in signal (either up or down) not associated with a biological response and 2) Drifting of the negative control well signal over the time course of the assay. Such artifacts are reduced using software algorithms including negative control correction (CeuticalSoft, Hudson, NY). However, the versatility of FDSS6000 hardware allows scientists to address addition artifacts, a more elegant solution than post assay data manipulation. In this study we compare the effects of cell plate shaking, pipettor fluid addition height, and pipettor fluid addition speed on reducing artifacts and improving assay response, using the FDSS6000
|  | Comparison of the No Wash Reagent Kits on the FDSS6000- Author: Hamamatsu Photonics, K.K.
- Source: Hamamatsu Photonics, K.K.
- Date: Mar-06
- Abstract: Abstract: Several commercially available 'No-Wash' reagent kits for measuring calcium mobilization using the FDSS6000 are now available. In some cases materials contained in a given kit may affect assay performance. Each kit offers advantages in terms of cost, performance, ease of use, availability, technical support, etc. We present examples showing the effect of kit choice on assay performance, in two separate studies.
|  | Multiplexing Calcium Mobilization and Membrane Potential Assays Using the FDSS6000.- Author: Hamamatsu Photonics, K.K.
- Source: Hamamatsu Photonics, K.K.
- Date: Mar-06
- Abstract: The FDSS6000 from Hamamatsu Photonics Systems uses Xenon lamps, enabling the use of fluorescent dyes ranging from UV to visible excitation wavelengths. FDSS6000 optics can read up to two excitation wavelengths and two emission wavelengths in the same assay. Current high throughput screening (HTS) strategies use only one dye per assay, such as Fura-2-AM, Fluo-3-AM, or Fluo-4-AM (calcium mobilization), or DisBAC2(3) (membrane potential). We present a novel multiplexing method for HTS Screening: Loading the same cells using both the UV dye Fura-2 AM and a no wash Membrane Potential Dye. Multiplexing assays measure orthogonal cell responses (calcium mobilization and membrane potential) using one volume of valuable library compound.
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