HMSiR – Fluorescent Probe for Live-Cell Super-Resolution Imaging
Live-Cell Super-Resolution Imaging of Microtubules in Velo Cells
Velo cells, in which HaloTag®-ß-tubulin was expressed, were stained by HMSiR-Halo. Images were obtained by STORM microscopy (N-Storm).
A: Averaged image. B: Super-resolution image. C: Localization precision measurements.
At first, antibodies were labeled with HMSiR, absorbed on the glass surface, and observed by STORM (N-STORM). Positions of molecules were determined repeatedly and their distribution was fitted by Gaussian distribution. Only signals over 150 photons were analyzed. Super-resolution imaging with HMSiR does not need thiols or oxygen scavengers and can be performed under physiological conditions. HaloTag® is a trademark of Promega Corporation.
Reaction of HMSiR
Fluorescence of Single HMSiR Molecule
HMSiR displays spontaneous blinking without high-power laser irradiation which normally is necessary for dSTORM observations (100 W cm-2 is sufficient instead of ~1 kW cm-2). Hence cell damage by laser irradiation can be minimized significantly.
- Imaging possible under physiological conditions
- HMSiR displays spontaneous blinking with low-power excitation light and without additives
- Overcoming the problems of dSTORM: performing super-resolution imaging without thiols, oxygen scavengers or high-power laser irradiation
- Minimize cell damage
- Super-resolution microscopy is a form of light microscopy. Due to the diffraction of light, the resolution of conventional light microscopy is limited. Super-resolution techniques allow images to be taken with a higher resolution than the diffraction limit providing many new insights in cellular biology. Direct stochastic optical reconstruction microscopy (dSTORM) is a super-resolution imaging technique that utilizes sequential activation and time-resolved localization of photoswitchable fluorophores to create high-resolution images, allowing spatial resolution of about 20 nm.
- The new released HMSiR series displays spontaneous blinking when it emits fluorescence. Unlike the emission of other probes - which show a constant fluorescence signal - this probe is flashing. Until now fluorophores did not blink by itself without the addition of additives. On the other hand, however, additives are interfering with live cell imaging. By using the HMSiR series super-resolution imaging is now possible without additives like thiols or oxygen scavengers. Furthermore, irradiation with high-power laser was necessary in previous dSTORM observations. This new probe enables live-cell super-resolution imaging with low-power excitation light retaining physiological conditions.
Live Cell Imaging under Physiological Conditions
Spontaneous Blinking Without High-Power Laser Irradiation
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|A202-01||HMSiR labeled Goat anti-mouse IgG (whole)||100 μg||896,00|
|A203-01||HMSiR labeled Goat anti-rat IgG (whole)||100 μg||896,00|
|A204-01||HMSiR labeled Goat anti-rabbit IgG (whole)||100 μg||896,00|
- All prices are in EURO excl. VAT and shipping. Only available in Europe.
- Uno SN. et al.: A spontaneously blinking fluorophore based on intramolecular spirocyclization for live-cell super-resolution imaging. Nat Chem. 681-689, (2014) Pubmed
- GORYO Fluorescent Probes Catalog (PDF)